Dapagliflozin attenuates endothelial cell pyroptosis and dysfunction induced by oxidized low-density lipoprotein / 中国组织工程研究

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.

Effect of Gualou Xiebai Banxiatang on Myocardial Microangiogenesis and HIF-1α/VEGF-related Pathways in Myocardial Ischemia Model Rats / 中国实验方剂学杂志

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.

Effect and Potential Mechanism of Inhibition of Long Non-coding RNA MALAT1 on Glycolipipotoxicity-induced Endothelial Cell Dysfunction / 中国循环杂志

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.

The effect of varied exercise intensity on antioxidant function, aortic endothelial function, and serum lipids in rats with non-alcoholic fatty liver disease / El efecto del ejercicio de intensidad variada sobre la función antioxidante, la función endotelial aórtica y los lípidos séricos en ratas con enfermedad del hígado graso no alcohólico

Abstract This study aimed to compare the effects of diet and exercise of different intensities on antioxidant function, aortic endothelial cell function and serum lipids in NAFLD (nonalcoholic fatty liver disease) rats. Fifty Sprague-Dawley (SD) rats (180-220g) were randomly divided into two experimental groups and fed either a standard rodent chow diet (CON; n=10) or a high-fat diet (HFD; n=40). After 16 weeks, the animals that received the HFD were randomly separated into a high-fat control group (HFC; n=10) or three exercise training groups: HFD and low-intensity exercise (LE; n=10), HFD and moderate-intensity exercise (ME; n=10), and HFD and incremental intensity exercise (IE; n=10). These experimental rats keep sedentary or trained for the next six weeks. A detection kit was used to detect nitric oxide synthase (NOs), nitric oxide (NO), malondialdehyde (MDA) and other markers of aortic oxidative stress. The expression levels of endothelial nitric oxide synthase (e-NOS) and endothelin-1 (ET-1) were detected by immunohistochemistry. TC, TG, and other lipid metabolism parameters were detected by an automatic analyzer. Exercise with different intensities could improve lipid metabolism, enhance antioxidant function, reduce MDA (P<0.01), increase NO (P<0.01), and improve the expression of e-NOS and ET-1 (P<0.01) protein levels in NAFLD rats. Decreased blood lipids were exhibited in all exercise groups. Notably, the moderate-intensity exercise demonstrated more effect on increasing glutathione (GSH) contents (P<0.01) and decreased the expression of ET-1 protein levels (P<0.01). The results showed that exercise at different intensities improved lipid metabolism and enhanced anti-oxidation function. Moderate exercise could improve the function of aortic endothelial cells.

Resumen Este estudio tuvo como objetivo comparar los efectos de la dieta y el ejercicio a diferentes intensidades sobre la función antioxidante, la función de las células endoteliales aórticas y los lípidos séricos en ratas NAFLD (con enfermedad del hígado graso no alcohólico) y alimentados con una dieta estándar para roedores (CON; n = 10) o con una dieta alta en grasas (HFD; n = 40). Después de 16 semanas, los animales que recibieron HFD se separaron aleatoriamente en un grupo de control alto en grasas (HFC; n=10) o tres grupos de entrenamiento físico: HFD y ejercicio de baja intensidad (LE; n=10), HFD y ejercicio de intensidad moderada (ME; n=10), y HFD y ejercicio de intensidad incremental (IE; n=10). Estas ratas experimentales se mantuvieron sedentarias o entrenadas durante las próximas seis semanas. Se utilizó un kit de detección para determinar óxido nítrico sintetasa (NO), óxido nítrico (NO), malondialdehído (MDA) y otros marcadores de estrés oxidativo aórtico. Los niveles de expresión de la óxido nítrico sintetasa endotelial (e-NOS) y endotelina-1 (ET-1) se detectaron mediante inmunohistoquímica. El analizador automático detectó TC, TG y otros parámetros del metabolismo de los lípidos. El ejercicio con diferente intensidad mejoró el metabolismo de los lípidos, mejoró la función antioxidante, redujo la MDA (P <0,01), aumentó el NO (P <0,01) y mejoró la expresión de los niveles de proteína e-NOS y ET-1 (P <0,01) en ratas NAFLD. Se observó una disminución de los lípidos en sangre en todos los grupos de ejercicio. En particular, el ejercicio de intensidad moderada demostró un mayor efecto en el aumento del contenido de glutatión (GSH) (P<0,01) y disminuyó la expresión de los niveles de proteína ET-1 (P<0,01). Los resultados mostraron que el ejercicio a diferentes intensidades mejoró el metabolismo de los lípidos y mejoró función antioxidante. El ejercicio moderado podría mejorar la función de las células endoteliales aórticas.

Clinical study on treatment of senile hypertension with Bushen-Yijing-Tiansui Decoction combined with levoclodipine besylate tablets / 国际中医中药杂志

Objective:To observe the therapeutic effect of Bushen-Yijing-Tiansui Decoction combined with levoclodipine besylate in the treatment of senile hypertension. Methods:A total of 150 elderly hypertensive patients admitted in our hospital from January 2019 to February 2020 were randomly divided into 2 groups by a random number table method, with 75 in each. Both groups were given basic symptomatic treatment of other comorbidities and concurrent health education. The control group received oral amlodipine besylate tablets, and the observation group received Bushen-Yijing-Tiansui Decoction on the basis of the control group. Both groups were treated for 4 weeks. The TCM syndromes were scored before and after treatment, manometer was used to measure blood pressure and heart rate, the serum ET-1 level was detected by Enzyme-Linked Immunosorbent Assay (ELISA), and the nitrate reductase method was used to detect serum NO levels, and adverse events occurred during treatment in the two groups were recorded. Results:The total effective rate of the observation group was 88.0% (66/75), and the control group was 73.3% (55/75), and the comparison difference in 2 groups was statistically significant ( χ2 =5.172, P=0.022). After treatment, the symptom scores of pain, palpitations, constipation, insomina, sore feeling on back and legs of the observation group were signigicantly lower than those in the control group ( t value were 5.814, 10.397, 12.094, 7.019, 6.121, all Ps<0.001). After treatment, heart rate [(79.60 ± 4.80) times/min vs. (84.30 ± 5.40) times/min, t=5.634], SBP [(144.8 ± 7.90) mmHg vs. (150.60 ± 7.90) mmHg, t=4.729], DBP [(78.80 ± 8.20) mmHg vs. (85.20 ± 9.10) mmHg, t=4.525] of the observation group were signigicantly lower than those of the control group ( P<0.01). After treatment, the serum ET-1 [(179.25 ± 30.45) μmol/L vs. (190.83 ± 30.89) μmol/L, t=2.312] of the observation group was signigicantly lower than that of the control group ( P<0.05), NO [(58.51 ± 8.78) μmol/L vs. (54.12 ± 9.03) μmol/L, t=3.019] of the observation group was signigicantly higher than that of the control group ( P<0.05). During treatment, the incidence of adverse events in the control group was 4.0% (3/75), and the observation group was 1.3% (1/75), and the two groups had no significant difference ( χ2 =1.027, P=0.311). Conclusion:Bushen-Yijing-Tiansui Decoction combined with levoclodipine besylate in the treatment of senile hypertension can relieve the clinical symptoms and blood pressure of patients, improve the function of vascular endothelial cells.

Effect of Yiqi Huoxue Recipe on Vascular Endothelial Function in Rats with Qi Deficiency and Blood Stasis Syndrome / 中国实验方剂学杂志

Objective: To explore the changes and benefits of vascular endothelial cell function in rats with qi deficiency and blood stasis syndrome, and the effect of Yiqi Huoxue recipe (YQHXF) of such changes. Method: Rats were randomly divided into blank control group, Qi deficiency and blood stasis group, and YQHXF high and low dose groups (5.54,2.77 g·kg-1). A small platform of water environment was used to make the rats stand for a long-term with irregular and incomplete sleep deprivation, 16 h per day for six weeks, so that both mentality and labor of rats were consumed to establish qi deficiency and blood stasis rat models. From the fifth week, intragastric administration was given for 2 weeks, until end of the experiment. Then levels of endothelin-1(ET-1), von willebrand factor (vWF), vascular cell adhesion molecule-1(VCAM-1), intercellular adhesion molecule (ICAM-1), P-selectin,interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in rat plasma were detected by enzyme-linked immunosorbent assay (ELISA) and nitric oxide (NO) was detected by nitrate reductase assay. Result:Compared with blank control group, rats in Qi deficiency and blood stasis group showed rough and dark hair, with significantly decreased body weight and pulse amplitude (PPPα were abnormally increased after sleep deprivation (PPPPPPPConclusion:Sleep deprivation can induce the formation of Qi deficiency and blood stasis syndrome in rats, and lead to vascular endothelial dysfunction. YQHXF has the function of protecting the vascular endothelium. It can improve the Qi deficiency and blood stasis symptoms in rats with Qi deficiency and blood stasis syndrome by regulating the secretion of vascular endothelial active substances, reducing cell adhesion and inhibiting inflammation.

The relationship between the level of serum ICAM, ALCAM, VCAM and PECAM-1 in elderly patients with type 2 diabetes mellitus combined with acute cerebral infarction and its clinical significance / 重庆医学

Objective To study relationship between the level of serum ICAM,ALCAM,VCAM and PECAM-1 in elderly patients with type 2 diabetes mellitus combined with acute cerebral infarction and its meaning.Methods From July 2014 to December 2015,60 patients with type 2 diabetes mellitus complicated with acute cerebral infarction were chosen as the research object.They were divided into three groups according to severity of atherosclerosis:no plaque in 11 cases,22 cases of stable plaques and unstable plaque 27 cases.40 cases with acute cerebral infarction patients and 40 cases of patients with type 2 diabetes were selected as a disease control;and other 40 healthy subjects were selected as healthy controls.ELISA were used to detect ICAM,ALCAM,VCAM and PECAM 1 level.Neural function evaluation was made by the U.S national institutes of health stroke scale (NIHSS).Results The ICAM,ALCAM,VCAM,PECAM 1 level in diabetic cerebral infarction group were higher than the other three groups,the difference was statistically significant (P＜0.05),while those of cerebral infarction group and diabetes group were higher than the control group,the difference was statistically significant (P＜0.05),and those of cerebral infarction group and diabetes group has no statistical significance (P＞0.05).NIHSS score of diabetic cerebral infarction patients was (7.39 ± 1.72),which was higher than that of patients with cerebral infarction group (5.33 ± 1.49),the difference was statistically significant (t=4.376,P =0.019,P＜0.05).The serum ICAM,ALCAM,VCAM,PECAM 1 level of diabetic cerebral infarction patients was a positively correlated with NIHSS score (r=0.559,P=0.007;r=0.619,P=0.000;r=0.421,P=0.018;r=0.451,P=0.007).With diabetic cerebral infarction is aggravating,the severity of carotid plaques in patients with serum ICAM,ALCAM,VCAM,PECAM-1 level is on the rise (P＜0.05).Conclusion ICAM,ALCAM,VCAM,PECAM-1 levels in peripheral blood serum of patients with type 2 diabetes mellitus complicated with acute cerebral infarction have an abnormal increase,and ICAM,ALCAM,VCAM,PECAM-1 level is closely related to the neurologic deficits and the severity of carotid artery plaque of type 2 diabetes mellitus complicated with acute cerebral infarction patients.

Effects of Different Constitutional Types of Coronary Heart Disease with Blood Stasis Syndrome among Patients with ApoE Gene Polymorphism on Vascular Endothelial Cell Function / 世界科学技术-中医药现代化

This article was aimed to study the distribution and effects of apolipoprotein E (ApoE) gene polymorphism of different constitutional types among coronary heart disease (CHD) patients with blood stasis syndrome on vascular endothelial cell (VEC) function. The whole gene sequencing method was used to identify genotypes of ApoE gene among 556 CHD patients with blood stasis syndrome, in order to analyze the relationship between polymorphism of ApoE gene and the level of VEC function. The results showed that the frequency of E3/3 genotype of each physical group was significantly lower than that of the healthy group; and the frequency of E3/4 genotype was significantly higher than that of the healthy group. In addition, the frequency of E3/3 genotype in qi deficiency constitution group was higher than that of blood stasis constitution group and yang deficiency constitution group; but the frequency of E3/4 genotype in blood stasis constitution group and yang deficiency constitution group was higher than that of the qi deficiency constitution group. The levels of ET and ET/NO in each of genotype groups of blood stasis constitution, yang deficiency constitution, qi deficiency constitution of CHD patients with blood stasis syndrome were higher than that of the healthy group (P < 0.05). In the genotype group of blood stasis constitution of CHD patients with blood stasis syndrome, the frequency of E3/4+E4/4 genotype ET was higher than that of other genotypes (P < 0.01). The levels of ET, NO, ET/NO in genotype groups of yang deficiency constitution and qi deficiency constitution of CHD patients with blood stasis syndrome were not significantly different. It was concluded that ApoE genotype E3/4, E3/3 may be the susceptible genotypes of blood stasis syndrome in CHD. There is a certain difference among different constitutional types. CHD with blood stasis syndrome patients, who are the constitution of blood stasis, carrying the ApoE gene polymorphism of E4 allele may have the function of increasing the ET level.

Study of Wenyang Huoxue Fang in Treating Patients with Acute Coronary Syndrome / 中国中医药信息杂志

Objective To study the effects of Wenyang Huoxue fang on patients with acute coronary syndrome in clinical presentation,inflammatory reactions,plasmin/plasminogen activators,endothelial cell injurys.Method Eighty cases of acute coronary syndrome were divided into treatment group and control group randomly,40 cases for each group.Both groups were treated with standard management(vasodilaters,cholesterol-lowering,antiplatelet) and the treatment group was given Wenyang Huoxue fang additionally for four weeks.TCM symptoms,effect of angina,TXB2,6-K-PGF,t-PA,PAI,IL-2,IL-6 were observed before and after treatment.Result Effective rate of angina in treatment group and control group was 80% and 65% respectively(P

The changes of endothelial cell function and coagulation-fibrinolysis system in maintenance hemodialysis patients with diabetic nephropathy / 中国实用内科杂志

Objective To find out the changes of endothelial cell function and coagulation-fibrinolysis system in maintenance hemodialysis(MHD)patients with diabetic nephropathy(DN).Methods The plasma levels of TM,PAI-1,TAT and PAP in 28 DN-MHD,34 non-DN MHD and 40 controls were measured by ELISA.Results (1)The levels of TM,PAI-1,TAT and PAP in both DN group and non-DN group were significantly higher than those in control group,P