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1.
Article in Chinese | WPRIM | ID: wpr-1021604

ABSTRACT

BACKGROUND:Graphene is the thinnest,strongest,and toughest type of two-dimensional new crystal material,demonstrating significant advantages in biomedical applications.Angiogenesis and vascularization of bone are key factors in tissue repair and regeneration,and are effective ways to address vascular and osteogenic issues. OBJECTIVE:To review the characteristics and mechanisms of graphene and its derivatives in promoting angiogenesis activity and vascularizing bone,in order to provide a reference for their clinical application in vascular tissue repair and regeneration. METHODS:Using a computer to search for relevant literature included in PubMed,ScienceDirect,CNKI,and Wanfang databases,the Chinese search terms were"grapheme","angiogenesis,vascularization","vascularized bone",and"endothelial cells",while the English search terms were"graphene""angiogenesis OR vascularization""vascularized bone""endothelial cells".After excluding literature unrelated to the topic of the article,according to the inclusion and exclusion criteria,62 articles were ultimately included for result analysis. RESULTS AND CONCLUSION:(1)At present,graphene oxide has been studied more and is the most widely used in graphene and its derivatives.(2)Graphene and its derivatives are suitable for heart,bone,nerve,and wound healing related diseases.(3)Graphene and its derivatives have excellent physical and chemical properties and biological properties,but they have potential cytotoxicity.We should pay attention to its biological safety in application.(4)The application of graphene and its derivatives requires further research to demonstrate the optimal size and concentration and measures to reduce toxicity.(5)On the cellular level,graphene and its derivatives can promote angiogenic activity by tip endothelial cell phenotype,mesenchymal stem cell adhesion and proliferation, and vascular smooth muscle cell growth.(6)On the molecular level,graphene and its derivatives can increase the expression of vascular endothelial growth factor,basic fibroblast growth factor,hepatocyte growth factor and activate reactive oxygen species/nitric oxide synthase/nitric oxide signaling pathway,lysophosphatilate R6/Hippo-YAP pathway,stromal cell-derived factor-1/vascular endothelial growth factor and ZEB 1/Notch1 pathway.(7)Grapheme oxide and graphene oxide-copper phosphorylated extracellular regulatory protein kinase and activated hypoxia-inducible factor-1,thereby promoting the up-regulation of vascular endothelial growth factor and bone morphogenetic protein-2 expression,and promoting angiogenesis and vascularized bone.(8)In summary,graphene and its derivatives,especially graphene oxide,have great application prospects in the repair and regeneration of vascularized tissues due to their excellent biological properties,good angiogenesis and vascularized bone ability.

2.
Article in Chinese | WPRIM | ID: wpr-1021652

ABSTRACT

BACKGROUND:Endothelial injury is one of the causes of cardiovascular diseases.Human induced pluripotent stem cells are easy to obtain,have strong differentiation ability,and have less exclusiveness,and their endothelial differentiated cells can be used as ideal cells for cardiovascular disease research. OBJECTIVE:To investigate the effect and mechanism of calycosin on endothelial differentiation of human induced pluripotent stem cells and to provide technical support for microvascular regeneration. METHODS:Human induced pluripotent stem cells were divided into control group and calycosin group(1.25,2.5 μg/mL),and growth factors were added to induce single-layer endothelial differentiation.After the induction of differentiation for 8 days,the positive rate of endothelial cell marker CD144 was detected by flow cytometry.Fluorescent expressions of CD144 and CD31 were detected by the immunofluorescence method.Lentivirus RNAi GFP puromycin was used to silence human-induced pluripotent stem cell Piezo1 mRNA followed by endothelial directed differentiation.After 8 days of differentiation,the positive rate of CD144 in differentiated cells was detected by flow cytometry.The mRNA expression levels of CD144,Piezo1 and MEK were detected by qPCR. RESULTS AND CONCLUSION:(1)Compared with the control group,the positive rate of CD144 was significantly increased in the 1.25 and 2.5 μg/mL calycosin groups(P<0.05).The expressions of CD144,Piezo1,and MEK mRNA were increased in the 2.5 μg/mL calycosin group(P<0.05).The fluorescence expressions of CD144(P<0.01)and CD31(P<0.001)were significantly increased in the 2.5 μg/mL calycosin group.(2)Compared with the shNT group,CD144 positive rate and CD144,Piezo1,MEK mRNA expressions were significantly increased in the shNT + calycosin 1.25,2.5 μg/mL groups(P<0.05).Compared with the shPiezo1 group,the positive rate of CD144 and mRNA expressions of CD144,Piezo1 and MEK had no significant changes in the shPiezo1+calycosin 1.25,2.5 μg/mL groups(P>0.05).(3)It is concluded that 2.5 μg/mL calycosin promotes the differentiation of human-induced pluripotent stem cells into endothelial lineages.Calycosin promotes the downstream MEK expression,thereby promoting the endothelial differentiation of human induced pluripotent stem cells by targeting the expression level of Piezo1.

3.
Article in Chinese | WPRIM | ID: wpr-1021722

ABSTRACT

BACKGROUND:Combining seed cells with 3D bioprinting technology enables the specific construction of various tissues and organs to meet the demands of tissue repair.However,further research is needed on the promotion of angiogenesis in damaged tissues. OBJECTIVE:By cultivating a 3D scaffold structure of methacrylated gelatin loaded with fibroblasts,obtaining the supernatant,and mixing it in different proportions with a complete culture medium to simulate the cellular microenvironment during tissue repair,this study aimed to explore the role of various cellular microenvironments in promoting angiogenesis in endothelial cells. METHODS:A methacrylated gelatin scaffold structure loaded with fibroblasts was prepared using an extrusion-based 3D bioprinting process.Hydrogel scaffold extract was prepared and mixed with a complete culture medium in ratios of 1:1,1:2,and 1:4 to obtain conditioned medium.Mouse embryonic fibroblasts BALB3T3 and human umbilical vein endothelial cells were co-cultured with complete medium(control group)and hydrogel scaffold extract,respectively.Cell proliferation was assessed using the CCK-8 assay and cell viability was analyzed using live/dead staining.Three kinds of conditioned medium and complete medium(control group)were used to co-culture with human umbilical vein endothelial cells for tube formulation assay,vascular genetic testing,and immunofluorescence staining of CD31. RESULTS AND CONCLUSION:(1)Scanning electron microscopy revealed that the methacrylated gelatin scaffold exhibited a porous structure,and rheological results demonstrated excellent mechanical properties of the hydrogel.CCK-8 assay and live/dead cell staining showed that the hydrogel scaffold extract had no obvious cytotoxicity.(2)Tube formulation assay indicated that the hydrogel showed the total length of cell tubules in 1:1 conditioned medium group was smaller than that in the control group(P<0.05).There were no statistical differences among the four groups in the number of vascular branches formed by endothelial cells(P>0.05).(3)qRT-PCR results showed that for vascular endothelial growth factor mRNA expression,the 1:2 conditioned medium group was lower than the 1:1 conditioned medium group on day 1(P<0.01).On day 3,the expression level of vascular endothelial growth factor in the 1:2 conditioned medium group was higher than that in the control group(P<0.01).On day 5,the cytokine expression level in the 1:2 conditioned medium group was significantly higher than that in the other three groups(P<0.01 or P<0.000 1).The expression in the 1:1 conditioned medium group was significantly lower than that in the other three groups(P<0.05 or P<0.01).On day 1,the expression level of basic fibroblast growth factor in the 1:1 conditioned medium group was significantly higher than that in the control group and 1:4 conditioned medium group(P<0.01,P<0.05).The expression was higher in the 1:2 conditioned medium group than that in the control group(P<0.05).On day 3,the expression levels of cytokines in the 1:4 conditioned medium group was higher than that in the control group(P<0.05).(4)On day 3,the expression of CD31 in the 1:2 conditioned medium group was higher than that in the control group and the 1:4 conditioned medium group(P<0.05).(5)The results indicate that the resulting conditioned media can simulate the microenvironment of vascular regeneration after tissue damage,promoting the vascularization process of endothelial cells.The best promotion of vascularization in endothelial cells was observed when the ratio of supernatant to complete culture medium was 1:2.

4.
Article in Chinese | WPRIM | ID: wpr-1021808

ABSTRACT

BACKGROUND:Heterotopic ossification of skeletal muscle is a clinically serious complication.For heterotopic ossification of skeletal muscles,the cells involved in the process of heterotopic ossification remain unclear. OBJECTIVE:To investigate the involvement of myocytes,fascia cells,and endothelial cells in the process of heterotopic ossification in skeletal muscle and to observe the cell origin of heterotopic ossification in skeletal muscle induced by bone morphogenetic protein 4. METHODS:Both C2C12 cells and the myotubes formed by the C2C12 cells in the induction medium were cultured,and 500 ng/mL bone morphogenetic protein 4 was added to the medium respectively,and whether the C2C12 cells and myotubes continued to proliferate within 10 days under the treatment were observed under a microscope.Myogenic cells(L6,derived from rats)and fibroblast-derived cells(derived from human)were co-cultured.After treatment with 500 ng/mL bone morphogenetic protein 4 and 10 ng/mL transforming growth factor-β,osteogenic and chondrogenic differentiation potential within 21 days were observed using Safranine O staining and Alcian blue staining.Using transgenic animal FVB/N-TgN(TIE2-LacZ)182Sato mice,15 μL of adeno-associated virus-bone morphogenetic protein 4(5×1010 PFU/mL)were implanted in the thigh muscle space of genetic mice for 10 and 14 days.X-gal staining was used to observe the formation of new blood vessel endothelium in the differentiated bone. RESULTS AND CONCLUSION:(1)Bone morphogenetic protein 4 caused myotube breakdown and increased C2C12 cell proliferation.Compared with other groups,the pure fibroblast-derived cell group had a higher area of positive alcian blue and safarin O staining(P<0.05)and a lower area of alkaline phosphatase staining(P<0.05),while the pure L6 group had a bigger area of alkaline phosphatase staining(P<0.05)but a smaller area of positive alcian blue and safarin O staining(P<0.05).(2)Transplantation of adeno-associated virus-bone morphogenetic protein 4-adsorbed gelatin sponge into FVB/N-TgN(TIE2-LacZ)182Sato mice resulted in heterotopic ossification.(3)X-gal staining results demonstrated that there was no obvious staining in chondrocytes and differentiated bones and Tie2+ endothelial cells did not participate in the formation of the alienated bone.(4)These findings verify that fibroblasts are the primary source of osteoblasts during the adeno-associated virus-bone morphogenetic protein 4-induced ectopic endochondral ossification in skeletal muscle,but myogenic cells are the main source of osteoblasts.Tie2+ endothelial cells might not be the cell source for cartilage and bone.

5.
Article in Chinese | WPRIM | ID: wpr-1021812

ABSTRACT

BACKGROUND:Increased homocysteine level induces apoptosis of human umbilical vein endothelial cells,but the mechanism remains unclear. OBJECTIVE:To investigate the role of hsa-circ-0001360 in human umbilical vein endothelial cell apoptosis induced by homocysteine. METHODS:In vitro cultured human umbilical vein endothelial cells were divided into control group,homocysteine group,interference control group,interference control + homocysteine group,hsa-circ-0001360 interference group,hsa-circ-0001360 + homocysteine interference group,overexpression control group,overexpression control + homocysteine group,hsa-circ-0001360 overexpression group and hsa-circ-0001360 + homocysteine overexpression group.All groups were treated with 100 μmol/L homocysteine.After 72 hours of intervention,the expressions of apoptosis-related proteins Bax,Bcl-2,and Caspase-3 were detected by western blot assay.The apoptotic rate was detected by flow cytometry.Quantitative real-time PCR was used to detect the expression of hsa-circ-0001360. RESULTS AND CONCLUSION:(1)Compared with the control group,the expression of Caspase-3 and Bax was significantly increased(P<0.01),and the expression of Bcl-2 was significantly decreased(P<0.01),and the apoptotic rate was significantly increased(P<0.01)in the homocysteine group.(2)Compared with control group,the expression of hsa-circ-0001360 was significantly increased in the homocysteine group(P<0.01).(3)The expression of hsa-circ-0001360 was significantly higher in the cytoplasm than that in the nucleus(P<0.01).(4)Compared with the interference control C group and interference control + homocysteine group,the expressions of Caspase-3 and Bax were significantly decreased(P<0.01),while the expression of Bcl-2 was significantly increased(P<0.01);the apoptotic rate was significantly decreased(P<0.01)in sh-hsa-circ-0001360 interference group and sh-hsa-circ-0001360 + homocysteine interference group.(5)Compared with overexpression control group and overexpression control + homocysteine group,the expressions of Caspase-3 and Bax were significantly increased(P<0.01),while the expression of Bcl-2 was significantly decreased(P<0.01);the apoptotic rate was significantly increased(P<0.01)in the hsa-circ-0001360 overexpression group and the hsa-circ-0001360 + homocysteine overexpression group.(6)In conclusion,hsa-circ-0001360 can promote the apoptosis of human umbilical vein endothelial cells induced by homocysteine.

6.
Article in Chinese | WPRIM | ID: wpr-1022542

ABSTRACT

Objective:We applied a hypoxia-induced model of human fetal retinal microvascular endothelial cell (RMEC) to study the effect of carbonic anhydrase 9 (CA9) on cell proliferation.Methods:The eyeballs of spontaneously aborted fetuses in Guangdong Women and Children's Hospital were obtained, and the retinas were isolated. RMEC was obtained by trypsin and collagenase two-step enzyme digestion, and endothelial cells were identified by CD34. The fetal RMEC and the purchased adult RMEC were cultured in normoxic and hypoxic incubators (1%O 2+5%CO 2+94%N 2), and the expression of CA9 was detected by qPCR and Western blot. After knocking down the CA9 by small interference RNA technique, the cell proliferation was detected by CCK-8 method, and the cell viability was detected by CCK-8 after adding CA9 inhibitor U-104. Results:The primary RMEC was extracted successfully. Immunofluorescence staining showed the percentage of CD34 positive cells in the third-generation cells was nearly 100%. The expression of CA9 mRNA in immature fetus and adult RMEC under hypoxia culture was higher than that under normoxic culture (fetal 1% O 2 group vs. fetal 21% O 2 group: 67.80±10.31 vs. 1.00±0.04, P<0.001; adult 1% O 2 group vs. adult 21% O 2 group: 1.72±0.22 vs. 1.00±0.02, P=0.014). Western blot analysis showed significantly increased expression of CA9 in the fetal RMEC exposed to hypoxia, which aligned with the expression of CA9 mRNA. When fetal RMEC was transfected with siCA9 20 nM, the knockdown rate of CA9 was 95% ( P<0.001). CCK-8 assay showed significantly lower proliferation of fetal RMEC cells in siCA9 group compared to siNC group (0.57±0.05 vs. 0.90±0.03, P<0.001), which was reflected by the OD value. With the addition of 100 μM CA9 inhibitor U-104, the viability of fetal RMEC in the treated groupwas significantly lower than that in the untreated group (99.16%±3.82% vs. 119.10% ±1.72%, P=0.002). Conclusions:The expression of CA9 differed between adult and preterm fetus in our hypoxia-induced RMEC model. Inhibiting CA9 can inhibit the proliferation of retinal microvascular endothelial cells of preterm fetus.

7.
Article in Chinese | WPRIM | ID: wpr-1022748

ABSTRACT

Objective To observe the effects of repeated intravitreal injections of ranibizumab and aflibercept on cor-neal morphology of patients with neovascular age-related macular degeneration(nAMD),diabetic macular edema(DME)or retinal vein obstruction(RVO).Methods In this prospective study,64 patients(64 eyes)who underwent therapy in the injection center of the Ophthalmology Department of our hospital from June 2021 to June 2022 were enrolled,including 19 nAMD patients,20 DME patients and 25 RVO patients.Among these patients,29 were treated with aflibercept(40 g·L-1)and 35 were treated with ranibizumab(10 g·L-1).Monocular injections were adopted for all patients,and 3+pro re nata(PRN)therapy was used.Confocal microscope was used for corneal nerve examination,and corneal endo-thelial microscope was used to measure corneal thickness(CT)and corneal endothelial cells.The CT,corneal endothelial cell density(ECD),coefficient of variation(CV),average cell size(ACS),proportion of hexagonal cells(Hex%),cor-neal nerve fiber length(CNFL),corneal nerve fiber density(CNFD)of patients with nAMD,DME and RVO after repeated intravitreal injections of anti-vascular endothelial growth factor(VEGF)drugs were compared,and those parameters at 1 month after injection of different anti-VEGF drugs were compared with the baseline.Results Before injection,ECD in the DME group was lower than that in the nAMD and RVO groups,and the ACS in the DME group was higher than that in the nAMD and RVO groups(all P<0.05).There was no significant difference in the other indexes among the three groups(all P>0.05).After 3 injections of anti-VEGF drugs,the ECD in the DME group was lower than that in the nAMD and RVO groups,the ACS in the DME group was higher than that in the nAMD and RVO groups,and the CNFL in the DME group was lower than that in the nAMD and RVO groups(all P<0.05).The ECD decreased compared with that before injection from the 2nd injection of aflibercept in the nAMD group(all P<0.05).Hex%decreased significantly after each injection compared with the baseline(all P<0.05).Other indexes have no significant differences from the baseline(all P>0.05).In the RVO group,ECD decreased from the 2nd ranibizumab injection compared with the baseline(all P<0.05).Conclu-sion Repeated intravitreal injections of anti-VEGF drugs can reduce the Hex%and ECD to a certain extent.After injec-tions,CNFL in the DME group is significantly lower than that in the nAMD and RVO groups.

8.
Article in Chinese | WPRIM | ID: wpr-1025103

ABSTRACT

Pyroptosis is a programmed mode of cell death.Activated caspase-1 can induce the occurrence of pyroptosis,promote the release of inflammatory factors,and trigger a violent inflammatory response.Depending on the type of caspase involved,pyroptosis can be divided into a caspase-1-mediated typical inflammasome pathway and a human caspase-4/5(or mouse caspase-11)-mediated atypical inflammasome pathway.In recent years,studies have found that pyroptosis is closely related to the occurrence,development,and prognosis of atherosclerosis.This article reviews the roles and mechanisms of endothelial cells,vascular smooth muscle cells,and macrophage cells in the occurrence and development of atherosclerosis,with the aim of promoting new ideas for research into the pathogenesis,diagnosis,and treatment of atherosclerosis

9.
Chinese Critical Care Medicine ; (12): 160-165, 2024.
Article in Chinese | WPRIM | ID: wpr-1025367

ABSTRACT

Objective:To observe the effect of lipopolysaccharide (LPS) induced conditioned medium of alveolar epithelial cells on the inflammatory response and cell damage of vascular endothelial cells, and explore its mechanism.Methods:The LPS induced type Ⅱ alveolar epithelial cells (A549) conditioned medium was used as a stimulus to induce human umbilical vein endothelial cells (HUVEC) damage. The cell counting kit-8 (CCK-8) was used to detect the effect of 0% (blank group), 12.5%, 25%, 50%, 75% and 100% A549 cell conditioned medium cultured for 6, 12, 24 and 48 hours on the cell viability of HUVEC. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)] and vasoactive substances [vascular endothelial growth factor (VEGF), endothelin-1 (ET-1)] in the supernatant. Phalloidin staining was used to observe the effects of A549 cells conditioned medium on cell morphology. The expressions of protein kinase B/nuclear factor-κB (AKT/NF-κB) pathway in HUVEC induced by conditioned medium was detected by Western blotting.Results:Compared with the blank group, A549 cells conditioned medium with concentrations of 12.5%, 25%, and 50% had no significant effects on cell viability of HUVEC after 6, 12, and 24 hours, but the activity of HUVEC decreased significantly after 48 hours. Therefore, 12.5%, 25%, 50% A549 cell conditioned medium stimulated for 24 hours was selected as the induction condition for follow-up experiments. Compared with the blank group, the level of IL-6 was significantly increased in 12.5% and 50% conditioned medium groups (ng/L: 2?438.95±64.89, 3?036.41±96.69 vs. 1?736.75±20.99, both P < 0.05), the level of TNF-α was significantly increased in 12.5% and 25% conditioned medium groups (ng/L: 174.08±11.09, 81.37±8.17 vs. 50.03±0.26, both P < 0.01), the levels of VEGF and ET-1 were significantly increased in 12.5%, 25% and 50% conditioned medium groups [VEGF (ng/L): 173.60±41.44, 192.49±12.38, 318.89±27.90 vs. 66.68±19.65; ET-1 (ng/L): 54.88±1.37, 36.69±0.29, 24.07±0.73 vs. 10.67±0.25, all P < 0.01]. Phalloidin staining showed that HUVEC induced by 25% A549 cells conditioned medium were irregular in shape, uneven in size, disordered in arrangement, widened in gap, dense and unclear in microfilament structure and serrated in cell membrane. Furthermore, the average fluorescence intensity of 25% conditioned medium group significantly increased compared to the blank group (67?205.60±3?430.40 vs. 56?272.67±7?650.95, P < 0.05). Western blotting showed that compared with the blank group, the expression of HUVEC cells phosphonated inhibitor α of NF-κB (p-IκBα) was significantly decreased in the 12.5%, 25%, and 50% conditioned medium groups (p-IκBα/IκBα: 0.38±0.08, 0.67±0.12, 0.31±0.07 vs. 1.00±0.00, all P < 0.01), the expressions of phosphonated-AKT (p-AKT) and VEGF were significantly increased (p-AKT/AKT: 1.50±0.18, 1.42±0.27, 1.61±0.14 vs. 1.00±0.00, VEGF/GAPDH: 1.37±0.10, 1.53±0.22, 1.40±0.12 vs. 1.00±0.00, all P < 0.05), the expression of phosphonated NF-κB p65 (p-P65) was significantly increased in the 25% conditioned medium group (p-P65/P65: 1.45±0.14 vs. 1.00±0.00, P < 0.05). Conclusion:LPS induced conditional culture medium of alveolar epithelial cells induced endothelial cell damage via activating AKT/NF-κB pathway.

10.
Chinese Circulation Journal ; (12): 185-193, 2024.
Article in Chinese | WPRIM | ID: wpr-1025452

ABSTRACT

Objectives:To investigate the effect of inhibition of long non-coding RNA(lnc RNA)in human metastasis associated lung adenocarcinoma transcript 1(MALAT1)on glycolipitoxicity-induced human umbilical vein endothelial cell dysfunction. Methods:Human umbilical vein endothelial cells were treated with glucose and palmitic acid in vitro to establish the glycolipitoxic endothelial cell models.Following groups were examined:control group,high-glucose and high-fat group,high-glucose and high-fat + non-targeting RAN control group,high-glucose and high-lipid+MALAT1 siRNA group,and high-glucose and high-lipid+MAPK1 siRNA group.RT-qPCR was used to detect the mRNA expression of MALAT1 and MAPK1.Western blot was used to detect the expression levels of autophagy,mitochondrial fusion division,apoptosis,and pathway-related proteins.Immunofluorescence confocal localization was used to detect the fluorescence colocalization of autophagy and lysosome-related proteins.The number of autophagolysosomes in endothelial cells was observed by transmission electron microscopy.Mitochondrial probe staining was used to detect mitochondrial morphology,immunofluorescence was used to detect intracellular reactive oxygen species(ROS)production,flow cytometry was used to detect the apoptosis of cells in each group,cell proliferation and scratch assays were used to detect the proliferation and migration ability of cells in different groups at different time points.The angiogenesis was quantified by counting the number of new blood vessels in each group. Results:Compared with the control group,the expression of lncRNA MALAT1 mRNA and the expression of phosphorylated mito-activated protein kinase 1(p-MAPK1)were upregulated(both P<0.05)and the expression of phosphorylated mammalian target protein(p-mTOR)was downregulated in the high-glucose and high-fat group and the high-sugar and high-fat control group(all P<0.01).Compared with the high-glucose and high-fat non-targeting RNA control group,the expressions of microtubule-associated protein 1A/1B-light chain 3(LC3)and p62 were downregulated(P<0.01,P<0.05),LC3 and lysosome-associated membrane protein 2(LAMP2)protein co-localized positive fluorescence particles were increased(both P<0.01),number of lysosomes were decreased,the expression of ROS was decreased(P<0.01),the expression level of mitochondrial fusion protein optic nerve atrophin 1(OPA1)was increased(P<0.05),the expressions of cleaved caspase-3 and BCL-2-related X protein(BAX)were decreased and BCL-2 was increased(all P<0.05),cell proliferation,migration,and tube-forming ability were increased(all P<0.01),and the expression of p-MAPK1 was decreased(P<0.05)and p-mTOR expression was increased(both P<0.05)in the high-glucose and high-lipid+si-MALAT1 group.Compared with the high-glucose and high-fat non-targeting RNA control group,the expression of p-MAPK1 in endothelial cells was decreased and the expression of p-mTOR was increased in the high-glucose and high-lipid+si-MAPK1 group(both P<0.01). Conclusions:Inhibition of lncRNA MALAT1 expression can reduce the level of mitophagy in glycolipidotoxic environments,reduce apoptosis of endothelial cells and improve endothelial cell function,which may be related to the regulation of MAPK1/mTOR signaling pathway.

11.
Article in Chinese | WPRIM | ID: wpr-1027416

ABSTRACT

Radiation-induced damage to vascular endothelium is a major complication of radiotherapy and a primary cause of morbidity and mortality in the population exposed to radiation. Ionizing radiation-induced cellular senescence serves as a critical factor in damage to vascular endothelial cells. Therefore, understanding the mechanisms of cellular senescence caused by senescence-associated secretory phenotype (SASP), as well as its role in ionizing radiation-induced damage to vascular endothelial cells, is significant for preventing and treating ionizing radiation-induced damage to vascular endothelial cells. In this study, the relationship between SASP-related premature senescence and this ionizing radiation-induced damage was explored from the following aspects: the mechanisms behind ionizing radiation-induced damage to vascular endothelial cells, ionizing radiation-induced cellular senescence, and the role of SASP-related premature senescence in the ionizing radiation-induced damage to vascular endothelial cells, as well as potential targets.

12.
Article in Chinese | WPRIM | ID: wpr-1030994

ABSTRACT

Objective @#To examine the role of LMO4 in the regulation of endothelial cell differentiation and angio- genesis in murine embryonic stem cells (mESC) .@*Methods @#Mouse Lmo4 cDNA was obtained from MEL cells by using the reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into the expression vector pFG to generate the pFLG ,in which contained Flk-1 promoter to drive Lmo4 expresses in only FLK-1 + cells.The mESC were transfected with pFG or pFLG plasmids and subsequently screened with geneticin ( G418) to produce cell clones. These cell clones were named mESC /pFG and mESC /pFLG ,respectively. The mESC /pFG and mESC /pFLG were cultured in the differentiation medium for either 4 days or 10 days to generate embryoid bodies (EB) .The 10-day embryoid bodies ( 10 d-EBs) carrying the pFG and pFLG vectors were subsequently stimulated to generate the blast-colony forming cells (BL-CFC) ,which indicated the presence of hemangioblasts.The endo- thelial cell sprouting analysis was performed by using 10 d-EBs.The expression of the interest genes was detected by using qualitative RT-PCR or Western blot analysis. @*Results @#The pFLG expression vector was successfully con- structed through PCR identification.The mESC /pFG and mESC /pFLG cells were obtained after transfected with the pFG or pFLG vectors and selected by G418.The cells spontaneously differentiate to generate EBs,in which some green fluoresce cells were present.Western blot analysis showed that a significant increase in LMO4 expression in both 4 d-EB and 10 d-EB when compared to mESC.BL-CFC analysis showed that the 4 d-EB/ pFLG had a higher cloning efficiency ( 7. 70% ± 1. 27% ) ,comparing with that of the 4 d-EB/ pFG ( 1. 15% ± 0. 48% ) ( P = 0. 021) .Quantitative RT-PCR results showed that the expression of Flk-1,C-kit,Tie-2 and Ve-cad genes in 10 d- EBs /pFLG increased more than 2-fold compared to 10 d-EBs /pFG.The endothelial cell sprouting analysis result showed a significant increase in the number and length of new blood vessels in 10 d-EB/ pFLG compared to 10 d- EB/ pFG (P<0. 05) .@*Conclusion @#Overexpression of LMO4 promotes hemangioblast differentiation from mESC, and benefits for endothelial cell differentiation and angiogenesis.

13.
Article in Chinese | WPRIM | ID: wpr-1031055

ABSTRACT

Background Vascular endothelial injury is an important pathogenic step of vibration-induced hand arm vibration disease (HAVD), and long-term vibration exposure can lead to vascular endothelial cell dysfunction and cell damage. Cell ferroptosis may be one of the important mechanisms of vibration-induced vascular endothelial cell injury and HAVD. Objective To explore whether vibration can induce changes in ferroptosis-related indicators in vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells (HUVEC) were divided into four vibrationgroups and two control groups. The vibration groups were exposed to an vibration setting of 125 Hz, 6.5 m·s−2 frequency band and for different durations: 1 d 2 h (total 1 d, 2 h per day), 1 d 4 h (total 1 d, 4 h per day), 2 d 2 h (total 2 d, 2 h per day), and 2 d 4 h (total 2 d, 4 h per day), respectively. All control groups were treated the same as the experimental groups except no vibration exposure. When the cells were 80% confluent, the control groups and the corresponding experimental groups were harvested at the same time. The effects of subgroup treatments on iron, reduced glutathione (GSH), and malondialdehyde (MDA) in HUVEC were detected with a cell ferrous colorimetric test kit, a reduced GSH colorimetric test kit, and a trace MDA test kit, respectively. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of ferroptosis-related genes acyl-CoA synthetase long chain family member 4 (ACSL4), tumor protein 53 (P53), recombinant human ferritin heavy chain (FTH1), and glutathione peroxidase 4 (GPX4). Western blot (WB) was used to detect the expression levels of ferroptosis-related proteins in HUVEC. Results Compared with the control groups, the vibration induced an increase in the iron content of HUVEC with a dose-response trend. Compared with the control groups, the reduced GSH content of HUVEC in the vibration group decreased with the increase of vibration time and frequency, and there was a dose-response trend. Compared with the control groups, the intracellular MDA content of HUVEC in the 1 d 2 h, 1 d 4 h, and 2 d 4 h vibration groups increased, and the MDA content in the 1 d 2 h and 1 d 4 h vibration group increased with time. The RT-PCR results showed that the mRNA expression levels of ACSL4 and P53 in the 1 d 4 h group increased compared with the 1 d 2 h group. Compared with the 2 d control group, the mRNA expression levels of ACSL4 in the 2 d 2 h vibration group and the 2 d 4 h vibration group increased, and the mRNA expression level of P53 in the 2 d 4 h vibration group increased. Compared with the 1 d control group, the mRNA expression levels of FTH1 and GPX4 in endothelial cells in the vibration 1 d 2 h group decreased. The WB results showed that compared with the control groups, the expression level of ferroptosis-related protein ACSL4 in endothelial cells increased in the vibration 1 d 2 h group; the expression levels of P53 in the 1 d 2 h and 2 d 4 h vibration groups increased; the expression levels of GPX4 decreased in the 1 d 4 h and 2 d 2 h vibration group, and the decrease was more obvious in the 2 d 2 h vibration group than in the 1 d 2 h vibration group; the above differences were statistically significant (P<0.05). Conclusion Vibration induces an increase in iron content, a decrease in GSH, and an increase in MDA in vascular endothelial cells in vitro, as well as mRNA and protein expressions of ferroptosis-related genes ACSL4, P53, FTH1, and GPX4.

14.
Article in Chinese | WPRIM | ID: wpr-1039315

ABSTRACT

Objective@#To explore the effect of adenoassociated virus sense transfection up-regulating the expressionlevel of the growth and differential factor 11 ( GDF11) in vivo on aortic injury in type 2 diabetic mellitus rats(T2DM).@*Methods@#Nine-week-old male SD rats were randomly selected to establish a T2DM model by usinghigh -sugar and high-fat chow plus small-dose streptozotocin (STZ) combined induction. Both normal rats and diabetic model rats were randomly divided into five groups: blank control group ( Control group) , negative virus control group ( NC group), GDF11 adeno-associated virus group ( GDFl1 group), diabetic group ( DM group), anddiabetic + GDF11 adeno-associated virus group ( DM + GDFl1 group). After 8 weeks of feeding, the serum con-centrations of insulin ( INS ), advanced glyeosylation end products ( AGES), recombinant growth transforming fac.tor 11 ( GDF11 ), total cholesterol (T-CH0 ), triglycerides (TG), low-density lipoproteins (LDL-C), high-density lipoproteins ( HDL-C), asymmetric dimethylarginine ( ADMA), and malondialdehyde ( MDA) were assayed inthe rats ; periodic acid-schiff stain( PAS stain) was used to observe the sites of glycogen deposition, and hematoxylin-eosin staining ( HE) was used to observe vascular damage. Scanning electron microscopy was used to observethe damage of vascular endothelial cells and vascular elastic fibers, and protein blotting and immunohistochemistrwere used to detect the expression levels of vascular injury-related proteins. Protein blotting and immunohistochemistry were used to detect the expression levels of vascular injury-related proteins. @*Results@#The biochemical indexes showed that the serum concentrations of AGES, T-CHO, TG, LDL-C and MDA were higher in the DM groupthan those in the Control group (P <0. 05), the concentrations of INS, GDF11, HDL-C and ADMA were signifi.cantly lower than those in the Control group (P <0. 05 ), and the concentrations of AGE'S and HDL-C were not sig.nificantly lower in the DM + GDF1l group compared with the DM group ( P <0. 05). HDL-C was not significantlydilerent from the DM group, and several other data were improved ( P<0. 05 ). Pathological staining suggestedthat PAS staining in the DM group suggested that glycogen particles deposited in the endothelium and subendotheli.um of the aorta, HE staining observed thickening of the aortic mesentery, endothelial cells and elastic fibers brokeolf in an irregular alignment , and electron microscopy observed endothelial damage in the vasculature and elastic fibers broke off in the DM group, and these changes attenuated in the DM + GDFl1 group. Protein blotting and im.munohistochemistry indicated that the expression of endothelial cell-associated proteins decreased in the DM groug( P <0. 05), and mesenchymal markers elevated in the DM group ( P <0. 05 ), these proteins were regressed inthe DM + GDFl1 group, and the difference was statistically significant (P<0.05).@*Conclusion@#Inereasing theexpression level of GDFl1 in vivo can improve aortic vascular injury caused by diabetes mellitus, which is inferredthat it may be related to the inhibition of endothelial mesenchymal transition to protect the function of vascular endo.thelial cells and thus improve vascular injury.

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Article in Chinese | WPRIM | ID: wpr-1039618

ABSTRACT

ObjectiveTo investigate the effect of Gualou Xiebai Banxiatang on cardiac function and myocardial histopathological changes in rats with ischemic myocardial injury, and to observe the effect of myocardial microvascular density (MVD), phosphatidylinositol 3-kinase (PI3K), mammalian target of rapamycin (mTOR), hypoxia-inducible factor-1 alpha (HIF-1α), and vascular endothelial growth factor (VEGF) signaling pathways on myocardial microangiogenesis. MethodSeventy male SD rats were randomly selected, with six rats in the normal group. The remaining rats were fed a high-fat diet and injected with isoproterenol hydrochloride (ISO,80 mg·kg-1·d-1, 2 d) to induce a hyperlipidemia-based ischemic heart disease model. After successful modeling, the rats were randomly divided into the model group, high, medium, and low dose groups of Gualou Xiebai Banxiatang, and the metoprolol group. The high, medium, and low dose groups of Gualou Xiebai Banxiatang were given Gualou Xiebai Banxiatang at 10.42, 5.21, 2.61 g·kg-1·d-1, respectively, while the metoprolol group was given metoprolol at 2.6 mg·kg-1·d-1. Both the normal and model groups were given an equivalent volume of physiological saline for 28 days. After the intervention, relevant tests were conducted, and serum was collected to measure heart function-related indicators. Hematoxylin-eosin (HE) and Masson staining were performed on ventricular tissue to observe pathological changes under a light microscope. Immunohistochemistry (IHC) was used to detect the positive expression of platelet endothelial cell adhesion molecule (CD31). Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of N-terminal pro-brain natriuretic peptide (NT-proBNP) and VEGF. Western blot was used to detect the protein expression levels of PI3K/mTOR/HIF-1α/VEGF. ResultCompared with the normal group, the model group showed significantly increased serum levels of LDH, CK, CK-MB, NT-proBNP, and VEGF (P<0.01), significantly increased collagen volume fraction (CVF) (P<0.01), significantly decreased MVD (P<0.01), and elevated protein expression levels of PI3K, mTOR, HIF-1α, and VEGF (P<0.05, P<0.01). Compared with the model group, the metoprolol group had significantly lower serum levels of LDH, CK, CK-MB, and NT-proBNP (P<0.01), significantly higher VEGF levels (P<0.01), significantly decreased CVF (P<0.01), significantly increased MVD (P<0.01), and significantly increased protein expression levels of PI3K, mTOR, and VEGF (P<0.01), with no statistically significant change in HIF-1α protein expression. Compared with the model group, the high and medium dose groups of Gualou Xiebai Banxiatang had decreased serum levels of LDH, CK, CK-MB, and NT-proBNP (P<0.05, P<0.01), increased VEGF levels (P<0.05, P<0.01), significantly reduced CVF (P<0.01), increased MVD (P<0.05, P<0.01), and significantly increased protein levels of PI3K, mTOR, HIF-1α, and VEGF (P<0.01). In the low dose group of Gualou Xiebai Banxiatang, compared with the model group, serum levels of LDH and NT-proBNP were decreased (P<0.05), VEGF was increased (P<0.05). Moreover, CVF was decreased (P<0.05), and the protein expression levels of PI3K, mTOR, HIF-1α, and VEGF were significantly increased (P<0.01). ConclusionGualou Xiebai Banxiatang can improve cardiac function, reduce myocardial pathological damage, enhance endothelial cell function, promote myocardial microvascular formation, and upregulate the expression of PI3K, mTOR, HIF-1α, and VEGF proteins in myocardial tissue in rats with ischemic myocardial injury.

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Article in Chinese | WPRIM | ID: wpr-1017808

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Objective To investigate the relationship between serum endothelial cell specific 1(endocan),deception receptor 3(DcR3),airway inflammation and clinical efficacy in children with bronchial asthma.Methods A total of 171 children with bronchial asthma who were admitted to the hospital from June 2020 to June 2022 were selected as the observation group,and 80 healthy children who underwent physical examina-tion in the hospital during the same period were selected as the control group.The levels of serum endocan,DcR3 levels and exhaled nitric oxide(FeNO)in the two groups were compared,and the correlation between the serum endocan,DcR3 levels and FeNO level in the observation group was analyzed.The levels of serum endocan and DcR3 were compared after individualized glucocorticoid treatment.The related factors affecting the clinical efficacy of the children were analyzed by univariate and multivariate Logistic regression analysis.Results The levels of serum endocan,DcR3 and FeNO in the observation group were higher than those in the control group(P<0.05).There was positive correlation between the levels of serum endocan,DcR3 and Fe-NO in the observation group(r=0.569,0.398,P<0.05).After treatment,the levels of serum endocan,DcR3 and FeNO in the observation group were lower than those before treatment(P<0.05).After treatment,there were 140 cases of children in the effective group,and 31 cases of children in the ineffective group.Univariate a-nalysis showed that there were statistical differences between the effective group and the ineffective group in the severity of disease,allergy history,endocan,DcR3 and FeNO levels(P<0.05).Multivariate Logistic re-gression analysis showed that severity of disease,allergy history,elevated serum levels of endocan and DcR3 were all independent risk factors affecting the clinical efficacy of children(P<0.05).Conclusion The levels of serum endocan and DcR3 are significantly related with airway inflammation in children with bronchial asth-ma,and their abnormally high levels could affect the clinical efficacy of glucocorticoid treatment in children with bronchial asthma.

17.
Journal of Clinical Hepatology ; (12): 533-538, 2024.
Article in Chinese | WPRIM | ID: wpr-1013133

ABSTRACT

ObjectiveTo investigate the role and mechanism of podoplanin (PDPN) in hepatic stellate cell (HSC) activation and liver fibrosis. MethodsLiver biopsy samples were collected from 75 patients with chronic hepatitis B who attended Department of Infectious Diseases, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, for the first time from September 2019 to June 2022, and RT-PCR and immunohistochemistry were used to measure the expression of PDPN in liver tissue of patients in different stages of liver fibrosis. A total of 12 male C57/BL6 mice were randomly divided into control group and model group. The mice in the model group were given intraperitoneal injection of 10% CCl4, and those in the control group were injected with an equal volume of olive oil, for 6 weeks. HE staining and Sirius Red staining were used to observe liver histopathological changes; primary mouse liver cells were separated to measure the mRNA expression of PDPN in various types of cells; primary mouse HSCs were treated with PDPN protein, followed by treatment with the NF-‍κB inhibitor BAY11-708, to measure the expression of inflammatory factors in HSCs induced by PDPN. The independent-samples t test was used for comparison of normally distributed continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. The Spearman correlation analysis was used to investigate data correlation. ResultsAs for the liver biopsy samples, there was a relatively low mRNA expression level of PDPN in normal liver, and there was a significant increase in the mRNA expression level of PDPN in liver tissue of stage S3 or S4 fibrosis (all P<0.001). Immunohistochemical staining showed that PDPN was mainly expressed in the fibrous septum and the hepatic sinusoid, and the PDPN-positive area in S4 liver tissue was significantly higher than that in S0 liver tissue (t=8.892, P=0.001). In normal mice, PDPN was mainly expressed in the hepatic sinusoid, and there was a significant increase in the expression of PDPN in CCl4 model mice (t=0.95, P<0.001), mainly in the fibrous septum. RT-PCR showed a significant increase in the mRNA expression of PDPN in the CCl4 model mice (t=11.25, P=0.002). Compared with hepatocytes, HSCs, Kupffer cells, and bile duct endothelial cells, hepatic sinusoidal endothelial cells showed a significantly high expression level of PDPN (F=20.56, P<0.001). Compared with the control group, the primary mouse HSCs treated by PDPN protein for 15 minutes showed significant increases in the mRNA expression levels of the inflammation-related factors TNFα, CCL3, CXCL1, and CXCR1 (all P<0.05), and there were significant reductions in the levels of these indicators after treatment with BAY11-7082 (all P<0.05). ConclusionThere is an increase in the expression of PDPN mainly in hepatic sinusoidal endothelial cells during liver fibrosis, and PDPN regulates HSC activation and promotes the progression of liver fibrosis via the NF-κB signaling pathway.

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Article in Chinese | WPRIM | ID: wpr-1013567

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Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm2 and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm2 and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.

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Article in Chinese | WPRIM | ID: wpr-1031582

ABSTRACT

【Objective】 To explore the plasma-activated medium (PAM) produced by low temperature plasma (LTP) on the proliferation and angiogenesis of human umbilical vein endothelial cells (HUVECs) so as to provide theoretical basis for the future use of PAM to promote wound healing and inhibit tumor angiogenesis. 【Methods】 HUVECs were selected as the in vitro research model. The PAM-containing medium after LTP treatment for different time points (0 s, 15 s, 30 s, 45 s, 60 s, and 75 s) was used for intervention. The influence of PAM on HUVECs viability was assessed using the MTT assay and cell cycle analysis. The effects of PAM on angiogenesis were examined through angiogenesis experiments. Intracellular levels of reactive oxygen species (ROS) were measured using fluorescence probes. A melanoma mouse model was established, and CD31 expression was detected by immunohistochemistry. 【Results】 As the treatment time increased, the intracellular levels of ROS also elevated. PAM derived from LTP exhibited a bidirectional effect on angiogenesis in HUVECs. Compared to the control group (0 s), low-dose treatments (15 s and 30 s) enhanced HUVECs viability, while high-dose treatments (45 s, 60 s, and 75 s) significantly decreased cell viability (P<0.05). The proportion of HUVECs in the S phase was significantly increased in the PAM-15 s and PAM-30 s groups, but markedly decreased in the PAM-45 s, PAM-60 s, and PAM-75 s groups, with statistically significant differences (P<0.05). The HUVECs tube formation ability was enhanced in the 15 s and 30 s PAM groups, but diminished in the PAM-45 s, PAM-60 s, and PAM-75 s groups, characterized by the decreased numbers of vascular nodes, intersections, meshes, and branching points (P<0.05). After PAM treatment in the melanoma mouse model, the control group exhibited widespread distribution of CD31 in tumor tissue, while the PAM-5 min and PAM-10 min groups displayed reduced distribution of CD31. 【Conclusion】 Short-term exposure to PAM enhances HUVECs proliferation and angiogenesis, whereas prolonged exposure suppresses cell viability and inhibits angiogenesis.

20.
Article in Chinese | WPRIM | ID: wpr-1021200

ABSTRACT

BACKGROUND:Dapagliflozin,an inhibitor of sodium-glucose cotransporter 2,can delay the progression of atherosclerosis by regulating glucose metabolism,inhibiting inflammation and improving endothelial cell function. OBJECTIVE:To study the effect of dapagliflozin on cell pyroptosis and endothelial dysfunction induced by oxidized low-density lipoprotein. METHODS:Human umbilical vein endothelial cells were divided into a control group(no intervention),a model group(treated with oxidized low-density lipoprotein for 24 hours),and a dapagliflozin group(treated with oxidized low-density lipoprotein + dapagliflozin for 24 hours).Endothelial cell proliferation activity was measured by cell counting kit-8 assay.The levels of intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 in cell supernatant were detected using ELISA.Nitric oxide level in the cells was detected by nitrate reductase assay.The pyroptosis rate and characteristics of endothelial cells were detected by Hoechst 33342/PI fluorescence co-staining and lactate dehydrogenase release assay.The protein expression levels of NLRP3,caspase-1,GSDMD,interleukin-1β,and interleukin-18 were detected by western blot assay. RESULTS AND CONCLUSION:(1)Oxidized low-density lipoprotein could cause pyroptosis and dysfunction of endothelial cells.(2)Compared with the control group,the level of nitric oxide and cell activity were decreased(P<0.05),while lactate dehydrogenase,intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 levels were significantly increased in the model group(P<0.05).Compared with the model group,cell activity and nitric oxide levels significantly increased(P<0.05),but lactate dehydrogenase,intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 levels were significantly diminished in the dapagliflozin group(P<0.05).(3)Compared with the model group,cell pyroptosis rate and the protein expression of pyroptosis factor NLRP3,caspase-1,GSDMD,interleukin-18 and interleukin-1β significantly reduced in the dapagliflozin group(P<0.05).(4)The results indicate that dapagliflozin inhibits oxidized low-density lipoprotein-induced endothelial pyroptosis and ameliorates endothelial cell dysfunction.

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