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+10 μg/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(collagen Ⅳ， 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。

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 μg/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 CⅣ 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 glucose-induced BM remodeling, and mitigating damage to iBRB.