【摘要】 目的:观察益景汤拮抗高糖诱导的体外血-视网膜内屏障(iBRB)模型基底膜(BM)损害及机制。方法:分离、培养大鼠内皮细胞(ECs)和视网膜微血管周细胞(RMPs)构建体外iBRB模型,随机分成低糖组、高糖组、米诺环素组、益景汤组,分别予25 mmol/L葡萄糖、60 mmol/L葡萄糖、60 mmol/L葡萄糖+10μg/mL米诺环素、60 mmol/L葡萄糖+10%益景汤含药血清干预,各组干预12 h后终止孵育。采用Western blot法检测各组BM相关蛋白[Ⅳ型胶原(collagenⅣ, CⅣ)、层黏连蛋白(laminin, LN)]及MMPs/TIMPs相关蛋白(MMP-2、MMP-3、MMP-9、TIMP-1、TIMP-2)的表达。结果:与低糖组相比,高糖组、米诺环素组、益景汤组CⅣ蛋白表达增加,高糖组、米诺环素组LN蛋白表达增加(均P<0.05)。益景汤及米诺环素能够抑制高糖诱导的CⅣ、LN蛋白表达增加,益景汤组、米诺环素组与高糖组相比具有差异(均P<0.05)。与低糖组相比,高糖组、米诺环素组MMP-2、MMP-3、MMP-9蛋白表达增加(均P<0.05)。益景汤能够抑制高糖诱导的MMP-2、MMP-3、MMP-9蛋白表达增加,益景汤组与高糖组相比具有差异(均P<0.05)。低糖组、高糖组、米诺环素组、益景汤组各组之间TIMP-1、TIMP-2表达未见明显差异(均P>0.05)。结论:益景汤可能通过调控MMP-2、MMP-3、MMP-9、CⅣ、LN的表达,干预高糖介导的BM重塑,抑制iBRB的损害,从而干预DR。更多还原

【Abstract】 ·AIM:To investigate the mechanism by which Yijing Decoction antagonist high glucose-induced damage to the basement membrane(BM)in an in vitro inner bloodretinal barrier(iBRB)model.·METHODS:Rat retinal microvascular pericytes(RMPs)and endothelial cells(ECs)were isolated and cultured to establish an in vitro iBRB model. The cells were randomly divided into four groups:low glucose group(LG),high glucose group(HG),minocycline group(MG)and Yijing Decoction group(YG). The LG group received 25 mmol/L glucose,the HG group received 60 mmol/L glucose,the MG group received 60 mmol/L glucose + 10 μ g/mL minocycline,and the YG group received 60 mmol/L glucose + 10% Yijing Decoction-containing serum.Incubation for each group were terminated after intervention for 12 h. Next,the Western blot analysis was performed to assess the protein expression of BM-related proteins,including collagen Ⅳ(CⅣ)and laminin(LN),as well as matrix metalloproteinase(MMPs)/tissue inhibitor of matrix metalloproteinases(TIMPs)such as MMP-2,MMP-3,MMP-9,TIMP-1,TIMP-2.·RESULTS:Compared to the LG group,the protein expressions of C Ⅳ increased in the HG,MG,and YG groups,as did LN in the HG and MG groups(all P<0.05).Both Yijing Decoction and minocycline effectively inhibited the elevated expression of CⅣ and LN induced by high glucose,and the difference between the YG,MG,and HG groups was statistically significant(all P < 0. 05).Futhermore,compared to the LG group,the protein expressions of MMP-2,MMP-3,and MMP-9 increased in the HG,MG,and YG groups(all P < 0. 05). Yijing Decoction specifically attenuated the high glucose-induced increase in MMP-2,MMP-3 and MMP-9 protein expression,and there were statistically significant differences between the YG and HG group(all P< 0.05).No significant difference were observed in the expressions of TIMP-1 and TIMP-2 among the LG,HG,MG,and YG groups(all P>0.05).·CONCLUSION:Yijing Decoction can potentially intervene in DR by modulating the protein expression of MMP-2,MMP-3,MMP-9,CⅣ,and LN,suppressing high glucoseinduced BM remodeling,and mitigating damage to iBRB.更多还原