miR-92a-3p promotes ox-LDL induced-apoptosis in HUVECs via targeting SIRT6 and activating MAPK signaling pathway

Atherosclerosis could be induced by multiple factors, including hypertension, hyperlipidemia, and smoking, and its pathogenesis has not been fully elucidated. MicroRNAs have been shown to possess great anti-atherosclerotic potential, but the precise function of miR-92a-3p in atherosclerosis and its potential molecular mechanism have not been well clarified. Flow cytometry assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol-3-ium bromide (MTT) assay were performed to evaluate effects of oxidized low-density lipoprotein (ox-LDL) on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs), respectively. Malondialdehyde and superoxide dismutase levels in cell lysate were assessed with biochemical kits. The expression levels of miR-92a-3p and Sirtuin6 (SIRT6) in HUVECs exposed to ox-LDL were estimated by real-time quantitative polymerase chain reaction (RT-qPCR). In addition, the protein levels of SIRT6, c-Jun N-terminal kinase (JNK), phosphorylation JNK (p-JNK), p38 mitogen activated protein kinase (p38 MAPK), and phosphorylation p38 MAPK (p-p38 MAPK) were measured by western blot assays. The relationship between miR-92a-3p and SIRT6 was confirmed by dual-luciferase reporter assay. Ox-LDL induced apoptosis and oxidative stress in HUVECs in concentration- and time-dependent manners. Conversely, miR-92a-3p silencing inhibited apoptosis and SIRT6 expression in HUVECs. The overexpression of miR-92a-3p enhanced apoptosis and phosphorylation levels of JNK and p38 MAPK as well as inhibited proliferation in ox-LDL-induced HUVECs. In addition, SIRT6 was a target of miR-92a-3p. miR-92a-3p negatively regulated SIRT6 expression in ox-LDL-induced HUVECs to activate MAPK signaling pathway in vitro. In summary, miR-92a-3p promoted HUVECs apoptosis and suppressed proliferation in ox-LDL-induced HUVECs by targeting SIRT6 expression and activating MAPK signaling pathway.

LncRNA XIST knockdown ameliorates oxidative low-density lipoprotein-induced endothelial cells injury by targeting miR-204-5p/TLR4

Oxidative low-density lipoprotein (ox-LDL)-induced endothelial cell injury is a key contributor toatherosclerosis development. However, the role and mechanism of long noncoding RNA X-inactive specifictranscript (XIST) in atherosclerosis remain largely unknown. The ox-LDL-induced human umbilical veinendothelial cells (HUVECs) injury was analyzed by cell viability, apoptosis, inflammatory cytokines secretionand oxidative stress. The expression levels of XIST, microRNA-204-5p (miR-204-5p) and toll-like receptor 4(TLR4) were detected by quantitative real-time polymerase chain reaction and western blot, respectively. Thetarget interaction between miR-204-5p and XIST or TLR4 was explored by bioinformatics analysis, luciferaseassay and RNA immunoprecipitation. The expression of XIST was enhanced in ox-LDL-treatedHUVECs. Knockdown of XIST attenuated ox-LDL-induced viability inhibition, apoptosis production,inflammatory response and oxidative stress in HUVECs. XIST was validated as a sponge of miR-204-5pand TLR4 acted as a target of miR-204-5p. Knockdown of miR-204-5p reversed silence of XISTmediated suppressive role in ox-LDL-induced injury. TLR4 alleviated miR-204-5p-mediated inhibitiveeffect on ox-LDL-induced injury. Moreover, XIST could regulate TLR4 expression by spongingmiR-204-5p. In conclusion, silence of XIST displayed a protective role in ox-LDL-induced injury inHUVECs by regulating miR-204-5p/TLR4 axis, providing a novel mechanism for understanding thepathogenesis of atherosclerosis.

Trimetazidine reduces formation of neutrophil extracellular traps via inhibiting autophagy / 中国药理学通报

Aim To study the effects of trimetazidine (TMZ)on theformationof neutrophil extracellular traps (NETs) induced by oxidized low-density lipoprotein (ox-LDL) in vitro and its relationship associated with cell autophagy. Methods The bone marrow neutrophils of mice were extracted by density gradient centrifugation,and NETs induction model was established using ox-LDL. Furthermore, TMZ, autophagy inhibitor LY294002 and autophagy inducer Rapamycin were added to disturb the induction of NETs induced by ox-LDL. The production of NETs marker MPO-DNA and the content of cfDNA/nets in the supernatantwere observed. Meanwhile, Western blot was employed to determine the protein expression of myeloperoxidase (MPO),beclin-l and LC3b in neutrophils. Results Treatment of ox-LDL to adhered neutrophils could lead to the formation of extracellular MPO-DNA and cfDNA/nets generation in a time-and concentration-dependent manner. The protein expression of LC3b,beclin-1 and MPO in neutrophils was up-regulated after treatedwith ox-LDL. TMZ pretreatment significantly inhibited NETs release and reduced the protein expression of LC3b,beclin-1 and MPO in neutrophils,which could be simulated by PI3K/AKT pathway blocker LY294002, while AKT/mTOR inhibitor rapamycin preconditioning did the opposite. Conclusions Ox-LDL-induces the formation of NETs in a time-and concentration-dependent manner. TMZ could down-regulate the level of autophagy and neutrophils, inhibit the induction of ox-LDL on NETs and reduce the release of NETs.

Up-regulated miR-106b inhibits ox-LDL-induced endothelial cell apoptosis in atherosclerosis

This research aimed to explore the molecular mechanism of microRNA (miR)-106b in cell apoptosis of atherosclerosis (AS). Human aortic endothelial cells (HAECs) were divided into control group, oxidized-low-density lipoproteins (ox-LDL) group, miR-106b NC+ox-LDL group, miR-106b mimics+ox-LDL group, miR-106b mimics+PTEN+ox-LDL group, and miR-106b mimics+empty+ox-LDL group. Real-time fluorescence quantitative polymerase chain reaction, cholecystokinin, TdT-mediated biotinylated nick end-labeling assay, luciferase reporter gene assay, and flow cytometry analysis were performed to determine the morphology, proliferation, and apoptosis in HSECs. Moreover, the levels of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), Bcl-2, p-P13K, and p-AKT in HAECs were detected by western blot. MiR-106b was down-regulated in ox-LDL-induced HAECs. PTEN was the target gene of miR-106b-5p. Overexpression of PTEN inhibited the anti-apoptotic effect of miR-106b. Compared with the control group, the proportion and number of HAECs apoptosis and Bax, caspase-3, and caspase-9 expression in ox-LDL and miR-106b mimics+PTEN+ox-LDL groups were significantly increased (all P<0.05). Moreover, the activity of HAECs and Bcl-2 were decreased significantly (all P<0.05). Overexpression of miR-106b in ox-LDL-induced AS inhibited endothelial cell apoptosis. Furthermore, miR-106b might activate the PI3K/AKT pathway by down-regulating the expression of PTEN in ox-LDL-induced HAECs.

Effects of moxibustion at 45 ℃ on blood lipoids and serum level of ox-LDL and NO in rats with hyperlipidemia / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion at different temperatures (38 ℃ and 45 ℃) on blood lipoids and serum level of oxidized low density lipoprotein (ox-LDL) and nitric oxide (NO) in rats with hyperlipidemia, and to explore the correlation between regulating blood fat and anti-oxidative stress and protection of vascular endothelium of moxibustion at 45 ℃.@*METHODS@#According to random number table, 60 SD rats were randomly divided into a normal group, a model group, a moxibustion at 38 ℃ group and a moxibustion at 45 ℃ group, 15 rats in each group. The rats in the normal group received no treatment; the rats in the remaining three groups were fed with high-fat diet for 8 weeks to prepare rat models of hyperlipidemia. After successful modeling, the rats in the model group received no treatment; the rats in the moxibustion at 38 ℃ group and moxibustion at 45 ℃ group were treated with moxibustion at "Shenque" (CV 8) and "Zusanli" (ST 36), and the temperature was controlled at (38±1) ℃ and (45±1) ℃, respectively. The moxibustion was given for 10 min at each acupoint, once every two days, and totally 4-week treatment was given. After treatment, the levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by using biochemical colorimetric method; the levels of ox-LDL and NO were measured by using ELISA method.@*RESULTS@#① Compared with the normal group, the levels of TC, TG and LDL-C were significantly increased in the model group (all 0.05). ② Compared with the normal group, the level of ox-LDL was increased but that of NO was decreased in the model group (both <0.01); compared with the model group and moxibustion at 38 ℃ group, the level of ox-LDL was decreased but that of NO was increased in the moxibustion at 45 ℃ group (<0.01, <0.05); compared with the model group, the level of ox-LDL was decreased but that of NO was increased in the moxibustion at 38 ℃ group (both <0.05).@*CONCLUSION@#Moxibustion at 45 ℃ has regulating effects on blood lipid in rats with hyperlipidemia, which can regulate blood lipid through various ways, such as anti-oxidative stress and protection of vascular endothelium.

MiR-590 Inhibits Endothelial Cell Apoptosis by Inactivating the TLR4/NF-κB Pathway in Atherosclerosis

PURPOSE: Previous study has well documented the anti-apoptotic effects of miR-590 on oxidized low-density lipoprotein (ox-LDL)-treated endothelial cells (ECs). However, the mechanism underlying the anti-apoptotic effects of miR-590 in ox-LDL-treated ECs remains to be further addressed. MATERIALS AND METHODS: ApoE(−/−) mice fed with a high-fat diet (HFD) and human aortic endothelial cells (HAECs) treated with ox-LDL were used as in vivo and in vitro models of atherosclerosis. The expressions of miR-590 and toll-like receptor 4 (TLR4) were detected by quantitative real-time PCR and Western blot, respectively. Atherosclerotic lesion analysis was performed using Evans blue and hematoxylin-eosin staining. Cell proliferation was assessed by MTT assay. Apoptosis was examined using flow cytometry analysis and Western blot analysis of Cleaved poly (ADP-ribose) polymerase (PARP) and Cleaved Caspase-3 levels. The effect of miR-590 on TLR4/nuclear factor kappa B (NF-κB) pathway was evaluated by Western blot. Binding between miR-590 and TLR4 was confirmed by luciferase reporter assay and Western blot. RESULTS: miR-590 was downregulated in the aorta tissues from HFD-fed apoE(−/−) mice and ox-LDL-treated HAECs. miR-590 overexpression inhibited atherosclerotic lesion in HFD-induced apoE(−/−) mice and promoted proliferation and inhibited apoptosis of ox-LDL-treated HAECs. Additionally, TLR4 was identified as a direct target of miR-590 in ox-LDL-treated HAECs. Moreover, anti-miR-590 reversed TLR4 knockdown-mediated promotion of cell proliferation and suppression of apoptosis in ox-LDL-treated HAECs. miR-590 overexpression suppressed the TLR4/NF-κB pathway, and inhibition of the TLR4/NF-κB pathway promoted cell proliferation and impeded apoptosis in ox-LDL-treated HAECs. CONCLUSION: miR-590 promoted proliferation and blocked ox-LDL-induced apoptosis in HAECs through inhibition of the TLR4/NF-κB pathway.

The role of lectin-like Ox-LDL receptor-1 on endothelial cell injury caused by human cytomegalovirus infection / 中华实验和临床病毒学杂志

Objective@#To investigate the role of lectin-like Ox-LDL receptor-1( LOX-1) in the activation and oxidative stress of cultured human umbilical vein endothelial cell(HUVEC) after human cytomegalovirus(HCMV) infection.@*Methods@#HUVEC were divided into four groups: HCMV, Control, Carrageenan, and HCMV+ Carrageenan. After HCMV AD169 infection, the supernatant of the culture was extracted, and cells were lysed. The levels of LOX-1 mRNA, intercellular adhesion molecule-1(ICAM-1) mRNA and vascular cellular adhesion molecule-1(VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxidum(NO) of the supernatant was detected by nitrate reductase method accordingly.@*Results@#24 h after infection, the mRNA expression of LOX-1, ICAM-1 and VCAM-1 in HUVEC of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly. The mRNA expression of LOX-1, ICAM-1 and VCAM-1 and the quantity of NO decreased after adding the LOX-1 inhibitor carrageenan. There was significant difference between groups(P<0.05).@*Conclusions@#HCMV may increase the mRNA expression of ICAM-1 and VCAM-1 and quantity of NO by upregulating the mRNA expresion of LOX-1, which may contribute to the formation of a therosclerosis(AS).

Effects of TET2-targeting miR-27b on oxidized low-density lipoprotein-induced inflammatory responses and apoptosis in endothelial cells / 中华微生物学和免疫学杂志

Objective@#To investigate the effects of miR-27b targeting ten-eleven translocation methylcytosine dioxygenase 2 (TET2) on oxidized low-density lipoprotein (ox-LDL) induced inflammatory responses and apoptosis in endothelial cells.@*Methods@#Double luciferase reporter gene analysis verified the targeting effect of miR-27b on TET2. Human umbilical vein endothelial cells (HUVECs) were induced by ox-LDL in vitro. Eight groups were set up including control group, ox-LDL group, ox-LDL+ anti-miR-con group, ox-LDL+ anti-miR-27b group, ox-LDL+ pcDNA group, ox-LDL+ pcDNA-TET2 group, anti-miR-27b+ si-con group and anti-miR-27b+ si-TET2 group. qRT-PCR was used to detect the expression of miR-27b and TET2 at mRNA level. Cell viability was detected by MTT assay. Cell apoptosis rate was detected by flow cytometry. Western blot was used to detect the expression of TET2, Cyclin D1 and caspase-3 at protein level.@*Results@#TET2 was the target gene of miR-27b. TET2 expression could be negatively regulated by miR-27b. ox-LDL increased the expression of miR-27b and reduced the expression of TET2 in HUVECs. The secretion of inflammatory factors and apoptosis rates of HUVECs in the control, ox-LDL+ anti-miR-27b, ox-LDL+ pcDNA-TET2 and anti-miR-27b+ si-con groups were significantly lower than those in the ox-LDL, ox-LDL+ anti-miR-con, ox-LDL+ pcDNA and anti-miR-27b+ si-TET2 groups, respectively (P<0.05).@*Conclusions@#miR-27b promoted ox-LDL-induced inflammatory responses and apoptosis in endothelial cells through down-regulating the expression of TET2.

Role of human βdefensin 2 on preventing oxidized low-density lipoprotein induced monocyte foaming / 中华传染病杂志

Objective To clarify the role of human β-defensin2 ( hBD2) on preventing oxidized low-density lipoprotein (OX-LDL) induced human leukemic monocyte (THP-1) foaming.Methods The monocyte foaming model was established using THP-1 cell induced by OX-LDL and the model was identified by oil red staining.The hBD2 was overexpressed on THP-1 cells by using lentivirus system and the effect of hBD 2 overexpression on THP-1 cell foaming induced by OX-LDL was detected.The levels of inflammatory factors including tumor necrosis factor (TNF)-α, interleukin (IL)-1βand IL-6 in cell supernatant of each group were detected by enzyme-linked immunosorbent assay ( ELISA).Differences between the groups were compared by using the t test.Results The gene transfection efficiency of the cells was close to 100%at 72 h after infection. The hBD2 protein levels were 0.122 ±0.024 in the control group, 0.123 ±0.022 in Lv-control infection group and 0.981 ±0.183 in Lv-hBD2 infection group; and the level in control group was statistically higher than that in hBD-2 infection group (t＝-3.175, P＝0.007).The relative levels of hBD2 mRNA at 72 h after virus infection were 0.131 ±0.021 in control group, 0.128 ±0.022 in Lv-control group and 1.001 ±0.105 in Lv-hBD2 infection group; and the level in control group was statistically higher than that in hBD-2 infection group (t＝-7.213, P＝0.003).The results of oil red staining showed that OX-LDL inducing THP-1 cells for 72 h could significantly induce lipid accumulation in cells.Overexpression of hBD2 could effectively inhibit lipid accumulation in THP-1 cells induced by OX-LDL.The expression of hBD2 mRNA in THP-1 group was significantly higher than that in THP-1+OX-LDL group (t＝3.237, P＝0.004); and the difference was also significant when comparing THP-1+Lv-hBD2+OX-LDL group with THP-1+OX-LDL group (t＝-6.021, P＝0.003).The level of hBD2 protein in THP-1 group was significantly higher than that in THP-1+OX-LDL group (t＝0.314, P＝0.006); and the difference was also significant when comparing THP-1+Lv-hBD2+OX-LDL group with THP-1+OX-LDL group (t＝-4.061,P＝0.007).The levels of TNF-α, IL-1βand IL-6 in the supernatant of THP-1 cells induced by OX-LDL for 72 h were significantly increased compared with those in THP-1group (t＝-3.825,-2.017 and -3.551, respectively; P＝0.007, 0.004 and 0.005, respectively). The levels of TNF-α, IL-1βand IL-6 in THP-1+Lv-hBD2+OX-LDL group were significantly lower than those in THP-1+OX-LDL group ( t＝4.132, 3.681, and 2.991, respectively; P＝0.003, 0.002, and 0.007, respectively).Conclusions hBD2 can effectively inhibit THP-1 foaming induced by OX-LDL, which may be related to its inhibition of inflammatory response.

microRNA-155 inhibits immuno-inflammatory reaction of foam cells by targeting MyD88 / 天津医药

Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.

Protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against oxidative injury / 中国中药杂志

The protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against ox-LDL-induced damage to human umbilical vein endothelial cells (HUVECs) was investigated by MTT assay, and the components were identified by using UPLC-Q-TOF-MS. According to the study, ethyl acetate extract and n-butanol extract could increase cell viability (P<0.01), while petroleum ether extract had no influence, and water extract could even inhibit the cell viability to some degree. Moreover, 32 compounds in four polar fractions were analyzed, including 31 quinones and their glycosides, and one rubiprasins C. Petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract contained 23, 32, 26, 15 compounds, respectively. According to cell experiments in vitro, active fractions were ethyl acetate extract and n-butanol extract. The results could provide scientific references for further studies on effective material basic of cRRR, and lay a foundation for studies on the relationship between efficacies and materials.

Effects of Gualou Xiebai Banxia decoction on blood lipid content, oxidative stress and ox-LDL/Lox-1 pathway in ApoE-/- mice / 中国中药杂志

To observe the functions of Gualou Xiebai Banxia decoction（GXBD） on regulating lipid metabolism, anti-oxidation, and interposing ox-LDL/Lox-1 pathway, and to explore its anti-atherosclerosis (AS) mechanisms. AS models were established by using 42 Apo-E-/- male mice with high fat diet. AS model mice were randomly divided into the model group, simvastatin group, and GXBD high and low dose groups. C57BL/6J male mice were used as the normal control group, n=10 and the treatment lasted for 8 weeks. The levels of TC, TG, LDL-C, HDL-C, SOD, MDA, GSH-px, and ox-LDL in blood serum were tested 24 h after the last administration. The changes of aortic tissues structure were observed by HE staining; the expression levels of Lox-1 protein and the expression levels of mRNA were detected by Western blot and PCR respectively.Results showed that the blood lipid levels and MDA, ox-LDL levels in blood serum of model group were significantly higher than those in the normal control group, but SOD, GSH-px levels were significantly lower than those in the normal control group, and the Lox-1 protein and mRNA expression levels were also significantly higher than those in the control group(P<0.05), namely aortic atherosclerosis lesions were obvious in model group.The levels of blood lipid and MDA, ox-LDL of GXBD high and low dose groups and simvastatin group were significantly lower than those in model group, while SOD, GSH-px levels were significantly higher than those in model group, and Lox-1 protein and mRNA expression levels were significantly lower than those in model group(P<0.05), namely the aortic atherosclerosis lesions were significantly relieved. The above results indicated that GXBD was capable of modulating blood lipid, anti-oxidation, and inhibiting the expression of Lox-1, and interposing ox-LDL/Lox-1 pathway in the AS model Apo-E-/- mice, which may be one of the mechanisms of anti-atherosclerosis.

microRNA-155 inhibits immuno-inflammatory reaction of foam cells by targeting MyD88 / 天津医药

Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.

Clinical Observation of Mailuoning Injection in Adjunctive Treatment of Acute Cerebral Infarction / 中国药房

OBJECTIVE:To observe clinical efficacy and safety of Mailuoning injection in adjunctive treatment of acute cere-bral infarction. METHODS:A total of 65 patients with acute cerebral infarction selected from neurology department of our hospital were divided into control group(32 cases)and observation group(33 cases)according to random number table. Control group was given conventional treatment. Observation group was additionally given Mailuoning injection 10 mL added into 0.9% sodium chlo-ride injection 250 mL intrnrenously,ivgtt,qd,on the basis of control group. Both group were treated for 15 d. Clinical efficacy as well as serum levels of ox-LDL,BNP and MMP-9,NIHSS score before and after treatment,the occurrence of ADR were com-pared between 2 groups. RESULTS:The response rate of observation group was 90.91%,which was significantly higher than 65.63%,with statistical significance(P0.05). After treatment,serum levels of ox-LDL,BNP and MMP-9,NIHSS score in 2 groups were all decreased significantly,and the observation group was significantly lower than the control group,with statis-tical significance (P0.05). CONCLU-SIONS:Mailuoning injection has significant therapeutic efficacy for acute cerebral infarction,can significantly reduce serum levels of ox-LDL,BNP and MMP-9,promotes neurological function and the recovery of patients with cerebral infarction with good safety.

The role of SR-BⅡ in the oxidized low density lipid induced foam cell formation promoted by porphyromonas gingivalis / 实用口腔医学杂志

Objective:To investigate the role of scavenger receptor BⅡ(SR-BⅡ) in the oxidized low density lipid(ox-LDL) induced foam cells formation promoted by porphyromonas gingivalis(P.g).Methods:Peritoneal macrophages were stimulated with P.g and ox-LDL,and then foam cells formation were checked.The expression of SR-BⅡ was detected by Western blot and real-time PCR.Next,siRNA targeting SR-BⅡ was used to detect the change of foam cells formation.Results:After being stimulated with P.g and ox-LDL,the foam cells formation was significantly increased.During the process of foam cells formation,P.g infection increased the expression of SR-BⅡ.And the knockdown of SR-BⅡ by siRNA significantly reduced the foam cells formation.Conclusion:P.g infection can increase the expression of SR-BⅡ and the regulation of SR-BⅡ expression can change the foam cells formation.

Puerarin attenuates the expressions ofHUVECs tissue factor and its inhibitor induced by ox-LDL / 基础医学与临床

Objective To investigate the effects of puerarin on the expression of human umbilical vein endothelial cells (HUVECs) tissue factor (TF) and tissue factor pathway inhibitor (TFPI) induced by oxidized low-density lipoprotein (ox-LDL).Methods After HUVECs were incubated with different concentrations of puerarin and 50 mg/L ox-LDL,the expression of TF and TFPI mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blot respectively.Results Compared with control,treatment with ox-LDL caused the augment of TF mRNA and protein expression (P<0.01),and the decrease of TFPI mRNA and protein expression.However,50,100,and 200 μmol/L puerarin blunted the augment of TF mRNA and protein expression and weakened the inhibition of TFPI mRNA and protein expression induced by ox-LDL(P<0.01).Conclusions Puerarin reduces HUVECs TF and TFPI mRNA and protein induced by ox-LDL.

Curcumin inhibits the apoptosis ofHUVECs induced by oxidized low density lipoprotein / 基础医学与临床

Objective To investigate the protective effect of curcumin on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL) and its possible mechanism.Methods Cultivated HUVECs were divided into six groups: control group, ox-LDL group, ox-LDL plus endoplasmic reticulum stress(ERS) inhibitor PBA group,curcumin group, ox-LDL plus curcumin group,ox-LDL plus curcumin plus PI3K inhibitor LY294002 group.Cell viabilities were evaluated by CCK-8 assays.The proportions of apoptotic cells were assessed by flow cytometry.The translocation of activating transcription factor 6 (ATF6) observed by laser confocal microscopy.Western blot was used to determine the expression of the ERS associated proteins:glucose-regulated protein 78(GRP78), protein kinase-like ER kinase(PERK), inositol-requiring kinase1(IRE-1) and the related pathways protein: LOX-1, AKT and phophorylated AKT.Results Compared with control group,increasedthe proportions of apoptotic cells(P<0.01),enhanced the expressions of ERS related proteins(P<0.01),promoted the transfer of ATF6 into the nucleus,as well as increased the expression of LOX-1(P<0.01)and decreased the expression of p-AKT(P<0.01) in the ox-LDL group;Compared with ox-LDL group,PBA inhibited ox-LDL-induced HUVECs apoptosis(P<0.01),curcumin inhibited ox-LDL-induced the expression of ERS associated protein and LOX-1(P<0.01), the nuclear translocation of ATF6, the apoptosis of HUVECs (P<0.01), and it also increased ox-LDL-induced down-regulation of p-AKT expression (P<0.01);LY294002 partially attenuated the inhibitory effect of curcumin on ERstress-related protein expression induced by ox-LDL(P<0.05).ConclusionsCurcumin can reduce ox-LDL induced apoptosis of HUVECs, its mechanism may be through the inhibition of LOX-1 expression and activation of AKT pathway to reduce ERS in cell.

Effects of hig hfat diet on reproductive func tion and the expression of Ox-LDL within testis in male mice / 医学研究生学报

Objective Lipid metabolism disorder can lead to male sterility, but its mechanism of affecting spermatogenesis and testis microenvironment remains unclear.This study aimed to investigate the effects of high fat diet on reproductive function and the expression of Ox-LDL within testis in male mice. Methods 16 C57BL/6 mice aged 8 weeks were divided into two groups by random number table method.High fat diet group was fed with high fat diet while normal fat diet group was fed with normal fat diet .At the end of 16 weeks, the levels of TG, TC, Ox-LDL and testosterone in serum were measured.The sperm concentration and motility from caudal epi-didymis were analyzed.The testis structure was observed by HE stai-ning.The localization and expression of Ox-LDL in testis were detec-ted by immunohistochemical technique. Results Compared with the normal diet group mice, the body weight (P0.05).The serum level of testosterone[(3.64 ±0.43)mg/L vs (0.40 ±0.14) mg/L, P<0.01], the sperm concentration[(9.95 ±0.75)106/mL vs (5.66 ±1.51)106/mL, P<0.01] and the sperm motility[(54.69 ±17.84)%vs (32.48 ±5.80)%, P<0.01] decreased in high fat diet group significantly.HE staining also showed that the amount of Leydig cells, spermatids and spermatozoons reduced obvi-ously in high fat diet group.Immunohistochemistry staining showed that Ox-LDL was mainly distributed around Leydig cells of mice tes-tis.The expression of Ox-LDL in high fat diet group increased significantly ( P<0.01) . Conclusion The expression of Ox-LDL in Leydig cells of high fat diet C57BL/6J mice increases significantly, which may inhibit testosterone biosynthesis and affect spermatogen-ic function.

Effect of tetramethylpyrazine injection on serum ion and β2-GPI/ox-LDL in patients with IgA nephropathy / 中国生化药物杂志

Objective To investigate the effect of tetramethylpyrazine injection on serum ion and β2-GPI/ox-LDL in patients with IgA nephropathy.Methods 58 IgA nephropathy patients were randomly divided into two gorups with 29 cases in each group.The control group were treated by benazepril and conventional treatment of dipyridamole, the experiment group were treated on the basis of the control group with tetramethylpyrazine injection.10 days of 1 courses, 2 courses of treatment.The serum ion,β2-GPI/ox-LDL and renal function related indexes were tested.Results Compared with before treatment, the serum inorganic phosphorus,β2-GPI/ox-LDL, Scr, BUN, CysC levels and 24 h urinary protein quantitation were lower( P<0.05), the blood calcium level were higher(P<0.05).Compared with the control group, the serum inorganic phosphorus,β2-GPI/ox-LDL, Scr, BUN, CysC levels and 24 h urinary protein quantitation were lower(P<0.05), the blood calcium level were higher(P<0.05), the clinical effect were higher (P<0.05).Conclusion Tetramethylpyrazine injection can effectively decrease the serum phosphorus, β2-GPI/ox-LDL, SCR, BUN, CysC levels and urinary protein amount of patients with IgA nephropathy, elevate blood calcium levels, significantly improve renal function and with less adverse reactions.

Effect of resveratrol on ROS production and PECAM-1 expression in ox-LDL-stimulated platelets / 中国药理学通报

Aim To investigate the effect of resveratrol on ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Methods The expression of PE-CAM-1 , Sirt1 and p38 MAPK phosphorylation in ox-LDL-stimulated platelets was determined by Western blot. The level of ROS was measured by immunofluo-rescence kit. Results ox-LDL induced platelet aggre-gation by 14%, whereas resveratrol inhibited platelet aggregation by 50%. Resveratrol decreased ROS level by 3 . 2 fold and completely suppressed PECAM-1 expression in ox-LDL-treated platelets. Resveratrol re-covered Sirt1 expression in ox-LDL-treated platelets. EX527 ( a Sirt1 inhibitor ) increased ROS level and PECAM-1 expression in ox-LDL-stimulated platelets. Meanwhile, resveratrol also suppressed p38MAPK phosphorylation induced by ox-LDL. Conclusion Resveratrol can inhibit platelet aggregation, decrease ROS production and PECAM-1 expression in ox-LDL-stimulated platelets. The mechanism maybe associated with recovery of Sirt1 expression. Moreover, resveratrol can decrease PECAM-1 expression, which may be linked to abolishing p38MAPK phosphorylation.