Effects Of MiR-21 And Rosuvastatin On Monocyte/macrophages

Part ? Role of miR-21 in monocyte/macrophages stimulated by ox-LDLObjectives:Previous studies have shown the expression of miR-21 was increased in the ox-LDL-treated peripheral monocytes and the atherosclerotic plaques in human.However,the role of miR-21 in monocyte/macrophages stimulated by ox-LDL is still unclear.Therefore,in this study the human monocyte/monocyte/macrophages were cultured in vitro and stimulated by ox-LDL.We aim to evaluate the effects of miR-21 on the monocyte/macrophages stimulated by ox-LDL.Methods:THP-1 cells were incubated with 160 nmol/L phorbol ester(PMA)to differentiate into monocyte/macrophages,monocyte/macrophages were treated with 50 mg/L ox-LDL,miR-21 inhibitors and negative control were transfected into monocyte/macrophages.The migration of monocyte/macrophages was detected by transwell.The cell supernatants were collected for quantification of inflammatory cytokine expression by ELISA.Oil red O staining for cytoplasmic lipid droplets in monocyte/macrophages.The expression of miR-21 was detected by real-time quantitative PCR.The expression level of dual specific phosphate-8(DUSP-8)and phosphorylation of MAPK pathway were detected by Western blotting.Results:miR-21 dificiency has no effect on ox-LDL induced macrophage cytokines production and lipid phagocytosis.Compared with the control group,the migration of monocyte/macrophages increased in the ox-LDL-treated group,the expression of miR-21 was increased,the expression level of DUSP-8 was decreased,the phosphorylation of JNK/SAPK and increased.Compared with the ox-LDL group,the expression of miR-21 was decreased in the miR-21 inhibitors group,the expression level of DUSP-8 was increased and the phosphorylation level of JNK/SAPK and p38 was decreased.Conclusion:These results demonstrate that Ox-LDL enhances the migration of monocyte/macrophages through JNK/SAPK and p38 signaling pathway by increasing miR-21 expression and suppressing DUSP8 expression.Part ? Effects of Rosuvastatin on migration of monocyte/macrophages stimulated by ox-LDLObjectives:Rosuvastatin has been shown to play pleiotropic effects in anti-atherosclerosis.However,its effect on migration of monocyte/macrophages is still unclear.We thus aimed to determine the influence of rosuvastatin on the migration of monocyte/macrophages in vitro.Methods:THP-1 cells were incubated with 160 nmol/L phorbol ester(PMA)to differentiate into monocyte/macrophages,monocyte/macrophages were treated with 50 mg/L ox-LDL and different concentrations of rosuvastatin,miR-21 mimics and negative control were transfected into monocyte/macrophages.The cell viability was detected by cell counting kit-8 The migration of monocyte/macrophages was detected by transwell.The expression of miR-21 was detected by real-time quantitative PCR.The expression level of dual specific phosphate-8(DUSP-8)and phosphorylation of MAPK pathway were detected by Western blotting.Results:Compared with ox-LDL group,the expression of miR-21 in statin group decreased,the expression of DUSP-8 was increased,the phosphorylation level of JNK/SAPK and p38 in MAPK pathway decreased,and the migration of macrophage was weakened.After transfection of miR-21 mimics,the expression of miR-21 in monocyte/macrophages increased,the expression of DUSP-8 was decreased,the phosphorylation level of JNK/SAPK and p38 increased,and macrophage migration was enhanced.Conclusion:Our study show that rosuvastatin can reduce the migration of monocyte/macrophages stimulated by ox-LDL through JNK/SAPK and p38 signaling pathway by suppressing miR-21 expression and increasing DUSP-8 expression.