Tanshinone IIA Reduces The Inflammatory Response Induced By Oxidized Low-density Lipoprotein By Down-regulating MiR-33 In THP-1 Macrophages

Part 1 Effects of ox-LDL and Tan on inflammatory factors in THP-1 macrophagesObjective: To investigate the concentration and time of ox-LDL and Tan stimulating THP-1 macrophages.Methods : THP-1 macrophages were treated with 0 ug/mL,10 ug/mL,20 ug/mL,40 ug/mL ox-LDL for 24 h or 20 ug/mL ox-LDL for different time(0 h,12 h,24 h,48 h).The expression levels of IL-1?,IL-6 and TNF-? were detected by ELISA.THP-1 macrophages were stimulated with 0 uM,5 uM,10 uM,20 uM Tan for 24 h or treated with 10 uM Tan for different time(0 h,12 h,24 h,48 h).The expression levels of IL-1?,IL-6 and TNF-? were observed.Results: The expression levels of IL-1?,IL-6 and TNF-? in THP-1 macrophages treated with ox-LDL were significantly increased(P<0.05)in a time-and concentration-dependent manner.The expression levels of IL-1?,IL-6 and TNF-? in THP-1 macrophages treated with ox-LDL at 20ug/mL for 24 h changed significantly,which was taken as the follow-up study object.Tan was treatment of THP-1 macrophages significantly reduced the expression levels of IL-1?,IL-6 and TNF-?(P<0.05),which showed a time-and concentration-dependent manner.The expression levels of(P<0.05)changed significantly at 24 h after treatment at 10 uM Tan,which was the follow-up study object.Conclusion: ox-LDL could promote the inflammatory response of THP-1 macrophages,while Tan could significantly inhibit the inflammatory response of THP-1 macrophages,which exerting anti-atherosclerosis effect,among which 10 uM Tan has a significant effect for 24 h.Part 2 Tan inhibits ox-LDL-induced inflammatory response by down-regulating the expression of mi R-33 in THP-1 macrophagesObjective: To explore the mechanism of Tan in stimulating THP-1 macrophages to secrete atherosclerotic cytokines induced by ox-LDL.Methods : Different concentrations of ox-LDL(0ug/m L,10ug/m L,20ug/m L or 40ug/m L)were used to stimulate THP-1 macrophages or 20ug/m L ox-LDL was used to stimulate THP-1 macrophages for 0h,12 h,24h and 48 h.The expression of mi R-33 was detected by q RT-PCR.Different concentrations of Tan(0u M,5u M,10 u M or 20 u M)was used to stimulate THP-1 macrophages induced by ox-LDL or 10 u M Tan was used to stimulate THP-1 macrophages for 0h,12 h,24h and 48 h,and observed the expression of mi R-33.Anti-NC and anti-mi R-33 were transfected into THP-1 macrophages,and q RT-PCR was used to observe the transfection effect.ELISA was used to detect the effect of down-regulation of mi R-33 on the expression of IL-1?,IL-6 and TNF-? in THP-1 macrophages.Results: The expression levels of mi R-33 in THP-1 macrophages treated with ox-LDL were significantly increased(P<0.05)in a time-and concentration-dependent manner.The expression levels of mi R-33 in THP-1 macrophages treated with Tan changed significantly in a time-and concentration-dependent manner.Compared with the anti-NC group,anti-mi R-33 significantly inhibited the expression of mi R-33(P<0.05),decreased the expression of IL-1?,IL-6 and TNF-? in THP-1 macrophages induced by ox-LDL(P < 0.05).Overexpression of mi R-33 significantly reversed the inhibition of Tan on the expression of IL-1?,IL-6 and TNF-? in THP-1 macrophages induced by ox-LDL.Conclusion: Tan inhibits the secretion of ox-LDL-induced proinflammatory cytokines by down-regulating the expression of mi R-33 in THP-1 macrophages,suggesting that Tan may play a role in atherosclerosis protection by targeting mi R-33 and reducing proinflammatory response.