1.Effect of fluvastatin on the expression of oxidized low-density lipoprotein-1 in human umbilical vein endothelial cells
Xiaolian ZHANG ; Deqian JIANG ; Shebing ZHANG
Chinese Journal of Tissue Engineering Research 2007;0(15):-
BACKGROUND:Oxidized low-density lipoprotein(ox-LDL) is recognized as the essential condition for atherosclerosis.As the ox-LDL-specific receptor,oxidized low-density lipoprotein receptor-1(LOX-1) involved in vascular inflammation and plaques develop-ment of the process.OBJECTIVE:To investigate the effect of fluvastatin on the expression of LOX-1 in human umbilical vein endothelial cells(HUVECs) induced by ox-LDL.DESIGN,TIME AND SETTING:The comparative observation was completed in the Medical Center of Second Xiangya Hospital,Central South University between August 2006 and May 2007.MATERIALS:Human umbilical vein endothelial cell line was purchased from the America ATCC Company.Fluvastatin original powder was supplied by Beijing Novartis Pharmaceutical Co.,Ltd.METHODS:HUVECs were incubated with:①Stimulation by ox-LDL with end concentration of 25,50,100 mg/L.②LOX-1 neutralizing antibody,and interfered with 50 mg/L ox-LDL.③Interfered with nuclear factor-?B(NF-?B) blockers pyrrolidine dithiocarbamate(PDTC),followed by 50 mg/L ox-LDL intervention.④Fluvastatin with concentration of 0.01,0.1,1 ?mol/L were used to interfere the cells,followed by 50 mg/L ox-LDL intervention.⑤There was an blank group as the control.MAIN OUTCOME MEASURES:The expression of LOX-1 mRNA level was detected by RT-PCR.RESULTS:The levels of mRNA of LOX-1 were increased after cells had beenwere incubated with different concertrations of ox-LDL,when compared with the control group.Within the dosage range of 25 to 50 mg/L,the above-mentioned indexes significantly changed in a concentration-dependent manner(P
2.Effect of epigallocatechin-3 gallate on cholesterol efflux in foam cells and its mechanism
Jianguo YIN ; Shebing ZHANG ; Daoquan PENG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2017;19(4):403-406
Objective To study the effect of epigallocatechin-3 gallate (EGCG) on cholesterol efflux in foam cells and its mechanism.Methods THP-1 cells were induced to differentiate into macrophages which were then transformed to foam cells.Foam cells were divided into 0 μmol/L EGCG group,10 μmol/L EGCG group,30 μmol/L EGCG group,and 100 μmol/L EGCG group (1.5 × 106 in each group).Their cholesterol content was measured with a cholesterol test kit,apoA-I-mediated cholesterol efflux was assayed with a liquid scintillation counter,expression of ATP-binding cassette A1 (ABCA1) was detected by RT-PCR and Western blot respectively.Results The ABCA1 mRNA and protein expression levels and cholesterol efflux were significantly higher while the cholesterol content was significantly lower in 10 μmol/L EGCG group,30 μmol/L EGCG group,and 100 μmol/L EGCG group than in 0 μmol/L EGCG group (7.04% ±0.21%,7.75%±0.17% and 8.53%±0.18% vs 3.37%±0.16%,P<0.01;419.33±19.75 mg/g,352.58± 14.23 mg/g and 312.62±17.45 mg/g vs 520.51 ±20.62 mg/g,P<0.01),and in 30 μmol/L EGCG group,100μmol/L EGCG group than in 10μmol/L EGCG group (P<0.05).Conclusion EGCG increases cholesterol efflux and decreases cholesterol content in foam cells by upregulating the transcription and expression of ABCA1.
3.Effect of Apolipoprotein AI on MCP?1 Plasma Levels,Monocyte Subset Proportions,and in Vitro CCR2 Expression
Jianguo YIN ; Shebing ZHANG ; Daoquan PENG
Journal of China Medical University 2017;46(6):501-504
Objective To explore effects of apoAI on MCP?1 levels in the plasma and the Ly6Chi monocyte proportion in the blood and spleen of atherosclerotic mice,as well as on CCR2 expression in vitro. Methods Sixteen male apoE?/?mice were fed with high cholesterol diet for 12 weeks. Mice were randomly divided into control and apoAI groups and were administered with PBS or apoAI(40 mg/kg),respectively,via tail vein on the 1st and 3rd day before sacrifice. Mice in both groups were administered LPS(25μg/mouse)via intraperitoneal injection 12 h before sacrifice. Plas?ma levels of MCP?1 were determined using ELISA,and the Ly6Chi monocyte proportion was analyzed using flow cytometry. In addition,human monocytic THP?1 cells were randomly divided into control and apoAI(10 mg/L)?treated groups,pretreated with corresponding intervention,and incubated with LPS(10μg/L). CCR2 expression levels were measured by Western blotting. Results Compared with the control treatment, apoAI treatment remarkably reduced MCP?1 levels in plasma and Ly6Chi monocyte proportion in the blood and spleen(P<0.01). Furthermore, apoAI treatment inhibited CCR2 expression in monocytes in vitro(P<0.05). Conclusion apoAI can reduce MCP?1 levels in plasma and the Ly6Chi monocyte proportion in the blood and spleen and can inhibit CCR2 expression in monocytes in vitro.
4.The expression levels of miR-21 and TGF-β1 in cardiac remodelin affer myocardial infarction
Fengping HE ; Xin XU ; Shebing ZHANG ; Baofeng CHEN ; Zhanzhong MA ; Shuguo YUAN ; Xiuyan HUANG ; Fenglian LIU ; Shiping FAN ; Dongnan WU
International Journal of Laboratory Medicine 2016;37(18):2513-2516
Objective To detect the change of exoression level of plasma microRNA‐21(miR‐21) and TGF‐β1 in cardiac remode‐lin affer acute myocardial infarction(AMI) of the pateins .Methods 200 pateints with AMI and 100 normal controls(age ,sex matched) were enrolled .Blood samples were obtained from the normal controls and patients with AMI on the 3 days ,7 days and 14 days .Real‐time PCR was developed to detect the expression of miR‐21 and TGF‐β1 in plasma .Results The expression of miR‐21 was significantly up‐regulation in the 3 days ,7 days and 14 days in MI group than that cntrol group ,0 .74 ± 0 .21 vs .2 .62 ± 0 .23 , vs .3 .67 ± 0 .25 ,vs .4 .13 ± 0 .27 up‐regulation in the 3 days ,7 days and 14 days in MI group than that cntrol group ,0 .98 ± 0 .18 vs .2 .35 ± 0 .24 ,vs .3 .67 ± 0 .25 ,vs .4 .13 ± 0 .27 ,P<0 .05 ,respectively .The expression of miR‐21 and TGF‐β1 were up‐regulation with the change of cardiac function .Positive relationship between miRNA‐21 expression and LVDd (r=0 .757 ,P<0 .05);Positive relationship between TGF‐β1 mRNA expression and LVDd(r=0 .701 ,P<0 .05) .Conclusion The expression of miR‐21 and TGF‐β1 were up‐regulation in cardiac remodelin affer AMI of the pateins ,which involved in regulation in cardiac remodelin affer AMI .
5.Apolipoprotein AⅠInhibits Lipid Retention and Apoptosis in Foam Cells via Induction of Autophagy
Jianguo YIN ; Shebing ZHANG ; Daoquan PENG
Journal of China Medical University 2018;47(1):17-21
Objective To observe the effects of apolipoprotein AⅠ (apoAⅠ) on autophagy,lipid retention,and apoptosis in foam cells, and to explore the anti-atherosclerotic mechanism of apoAⅠ. Methods Macrophages derived from THP-1 cells were randomly divided into the control group,the 3-MA+apoAⅠ group,and the apoAⅠ group. Each group was administered oxidized low-density lipoprotein for 36 hours,then lipid droplets and autophagosomes were observed and cellular cholesterol content was quantified. LC3 and Beclin-1 expression was examined by western blot and the apoptotic ratio determined using flow cytometry. Results Compared with the results of the control group,apoAⅠ treatment inhibited lipid retention and apoptosis in foam cells,decreased cellular cholesterol content,and up-regulated the expression of LC3 and Beclin-1 (P < 0.01). Administration of 3-MA abrogated the effects of apoAⅠ (P < 0.05). Conclusion apoAⅠ inhibits lipid retention and apoptosis in foam cells by inducing autophagy.