中国中医眼科杂志2026，Vol.36Issue(9)：801-808,8.DOI:10.13444/j.cnki.zgzyykzz.2026.09.001
从TGF-β/Smads通路探讨活血散结方对兔RPE细胞EMT的调控机制
Exploring the Regulatory Mechanism of Huoxue Sanjie Formula on EMT in Rabbit RPE Cells via the TGF-β/Smads Pathway
摘要
Abstract
OBJECTIVE To investigate the mechanism by which Huoxue Sanjie Formula regulates epithelial-mesenchymal transition(EMT)of retinal pigment epithelium(RPE)cells through the transforming growth factor-β(TGF-β)/Sma and Mad-related protein(Smad)signaling pathway.METHODS Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was employed to determine the concentrations of Notoginsenoside R1 and Tanshinone ⅡA in the drug-containing serum of Huoxue Sanjie Formula,and to screen the optimal concentration of drug-containing serum and the optimal dose of TGF-β type Ⅱ receptor degradation inducer 1(ITD-1).Primary rabbit RPE cells were isolated and routinely cultured to the third passage,then randomly divided into control group(CG),model group(MG),blank serum group(BSG),Huoxue Sanjie Formula medicine-containing serum group(HXSJ),and positive medicine group(ITD-1).Except for the CG group,all groups were treated with 10 ng/mL TGF-β1 for 48 h to establish an in vitro EMT model of RPE cells,followed by corresponding interventions.After 48 h,the expression of EMT marker proteins(E-cadherin,ZO-1,N-cadherin,α-SMA)was detected by enzyme-linked immunosorbent assay(ELISA);The expression of TGF-β/Smads pathway-related proteins(Smad2,Smad3,p-Smad2,p-Smad3,TGF-β R2)was detected by Western Blot,and the mRNA expression of Smad2 and Smad3 was detected by real-time quantitative PCR(RT-qPCR).RESULTS(1)Concentrations of active components:The concentrations of Notoginsenoside R1 and Tanshinone ⅡA in the medicine-containing serum of each group were within the linear range.(2)Intervention conditions:A 20％concentration of medicine-containing serum and a concentration of 3.44 mg/L ITD-1 were selected as the optimal intervention conditions.(3)EMT marker proteins:The protein expressions of E-cadherin,ZO-1,N-cadherin,and α-SMA in rabbit RPE cells of the MG group were significantly higher than those in the CG group,while those in the BSG group were significantly lower than those in the MG group;The expressions of E-cadherin and ZO-1 in the HXSJ and ITD-1 groups were significantly higher than those in the MG group,whereas the expressions of N-cadherin and α-SMA were significantly lower than those in the MG group,the differences were all statistically significant(all P＜0.05).(4)TGF-β/Smads pathway-related proteins:The protein expressions of Smad2,Smad3,p-Smad2,p-Smad3,and TGF-β R2 in RPE cells of the MG group were significantly higher than those in the CG group;Those in the BSG group were significantly lower than those in the MG group;and those in the HXSJ and ITD-1 groups were significantly lower than those in the MG group,the differences were all statistically significant(all P＜0.05).(5)mRNA expression levels:The mRNA expressions of Smad2 and Smad3 in RPE cells of the MG group were significantly higher than those in the CG group;Those in the BSG group were significantly lower than those in the MG group,and those in the HXSJ and ITD-1 groups were significantly lower than those in the MG group,the differences were all statistically significant(all P＜0.05).CONCLUSIONS Huoxue Sanjie Formula may inhibit the occurrence and development of EMT in rabbit RPE cells through the TGF-β/Smads signaling pathway.关键词
活血散结方/视网膜色素上皮细胞/上皮-间充质转化/TGF-β/Smads/增殖性玻璃体视网膜病变Key words
Huoxue Sanjie Formula/retinal pigment epithelial cells/epithelial-mesenchymal transition/TGF-β/Smads/proliferative vitreoretinopathy分类
医药卫生引用本文复制引用
戴金辉,彭俊,周晓容,白海霞,彭清华,陈向东,吴权龙..从TGF-β/Smads通路探讨活血散结方对兔RPE细胞EMT的调控机制[J].中国中医眼科杂志,2026,36(9):801-808,8.基金项目
湖南中医药大学校级重点项目(Z2023XJZD04) （Z2023XJZD04）
