Influence of ox-LDL on migration function of THP-1 macrophages and microRNA21 expression and MAPK pathway phosphorylation / 心血管康复医学杂志

Objective :To explore influence of oxidized low density lipoprotein (ox-LDL ) on migration function of THP-1 macrophages ,expression of microRNA 21 (miR-21) and mitogen-activated protein kinase (MAPK) path-way.Methods :Phorbol myristate acetate (PMA) of 160nmol/L was used to induce THP-1 cells to differentiate into macrophages.According to application of ox-LDL treatment and liposomes-mediated miR-21 inhibitor transfecting THP-1 macrophages (transfection for short) or not ,THP-1 macrophages were divided into blank control group (re-ceived neither ox-LDL treatment nor transfection ) ,ox-LDL group (received 50mg/L ox-LDL treatment without transfection) ,miR-21 inhibitor group (received 50mg/L ox-LDL treatment after transfection ) and miR-21 inhibitor negative-control group (received 50mg/L ox-LDL treatment after negative-control transfection ).THP-1 macro-phage migration number was measured by transwell method ,miR-21 expression was measured by real-time quantita-tive PCR ,and expression of dual specific phosphate 8 (DUSP-8) and phosphorylation level of MAPK pathway were measured by Western-blot method .Results :Compared with blank control group ,there were significant rise in mi-gration number of THP-1 macrophages [(74.10 ± 15.10) vs.(184.10 ± 26.28)] ,miR-21 expression [(1.00 ± 0.21) vs.(2.02 ± 0.27)] and phosphorylation levels of JNK and P38 protein ,and significant reduction in expression of DUSP-8 protein in ox-LDL group ,P＜0.01 all.Compared with ox-LDL group ,there were significant reductions in migration number of THP-1 macrophages [ (184.10 ± 26.28) vs.(58.50 ± 10.24)] ,miR-21 expression [ (2.02 ± 0.27) vs.(0.66 ± 0.16)] and phosphorylation levels of JNK and P38 protein ,and significant rise in expression of DUSP-8 protein in ox-LDL group , P＜0. 01 all .Conclusion : Ox-LDL enhances migration function of macrophages , which may be related to its effects of upregulating miR-21 expression ,enhancing phosphorylation of JNK and P38 protein of MAPK pathway and reducing DUSP-8 expression .

Lipid-lowering effect of seven traditional Chinese medicine monomers in zebrafish system / 生理学报

The present study aimed to study lipid-lowering effect of seven traditional Chinese medicine monomers in zebrafish system. Zebrafish were fed with high fat diet to establish a hyperlipemia model, then fasted and bathed with seven traditional Chinese medicine monomers stigmasterol, triacontanol, chrysophanol, vanillic acid, shikimic acid, polydatin and oleanolic acid respectively. The oil red O staining was used to detect the blood lipids of zebrafish. Serum total cholesterol and triglyceride levels were detected to validate the lipid-lowering effect. The result showed that a zebrafish model of hyperlipemia could be established by feeding larvae zebrafish with high fat diet. Among the seven traditional Chinese medicine monomers, chrysophanol had lipid-lowering effect. Chrysophanol significantly reduced serum total cholesterol and triglyceride levels in adult zebrafish fed with high fat diet. Chrysophanol accelerated peristalsis frequency of zebrafish intestine and the excretion of high fat food. It is concluded that chrysophanol has lipid- lowering effect in zebrafish, and the mechanism of the effect may be due to the roles of chrysophanol in reducing lipid absorption from gastrointestinal tract and accelerating the excretion of food.

Effects and mechanism of visfatin on MMP-9 in macrophages / 上海第二医科大学学报

Objective To investigate the effects and mechanism of visfatin on matrix metalloproteinases-9(MMP-9)expression and invasive activity in macrophages.Methods THP-1 monocytes were induced into macrophages.To investigate the effects of visfatin on MMP-9,cells were divided into 2 groups:①macrophages+visfatin 12 h;②macrophages+visfatin 24 h.The concentrations of visfatin in each group were:0(control),50,100,200,400 ng/mL.MMP-9 mRNA and protein expression were analysed by RT-PCR and Western blotting,and MMP-9 invasive activity was assayed by gelatin zymography.To investigate the mechanism of visfatin on MMP-9,cells were divided into 5 groups:①macrophages without stimulation(control);②macrophages pretreated with MAPK p38,ERK1/2,JNK pathway inhibitor for 1 h,then stimulated with visfatin(200 ng/mL)for 24 h;③macrophages pretreated with retinoid X receptors(RXR)nature ligand or peroxisome proliferators-activated receptor ?(PPAR?)natural/synthetic ligand for 1 h,then stimulated with visfatin(200 ng/mL)for 24 h;④macrophages stimulated with visfatin(200 ng/mL)for 24 h;⑤macophages+visfatin(200 ng/mL)for different time(5,10,15,30,60 min).MMP-9 expression,PPAR? expression,and the effect of visfatin on MAPK phosphorylation were analysed by Western blotting.Results Visfatin not only significantly enhanced MMP-9 mRNA and protein expression in macrophages(P