基于Notch通路研究活血散结方对TGF-β诱导的视网膜色素上皮细胞上皮-间充质转化的影响

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中文摘要：目的:通过视网膜色素上皮细胞(RPE)细胞骨架的形态学变化和细胞特征蛋白表达观察活血散结方对转化生长因子-β(TGF-β)诱导的RPE发生上皮-间充质转化(EMT)的影响,并从Notch信号通路转导后激活相关蛋白表达来明确活血散结方的作用机制.方法:体外提取原代RPE进行传代培养,将培养的RPE根据不同干预措施分为空白对照组、家兔血清组、TGF-β组、TGF-β+含药血清组、TGF-β+SB431542组、TGF-β+LY411575组;运用免疫荧光法观察活血散结方含药血清对TGF-β诱导的RPE的EMT,细胞骨架的形态学变化及ZO-1、α-SMA的表达;运用RT-PCR和Western Blot检测CBF-1、NICD1、Hes-1 mRNA和蛋白表达情况.结果:TGF-β+含药血清组RPE与TGF-β组比较,细胞骨架形态学纤维化明显减少,F-actin显著减少(P＜0.01),细胞排列整齐;上皮细胞特征蛋白ZO-1表达增加、间质细胞特征蛋白α-SMA的表达减少(P＜0.01),Notch信号通路涉及的CBF-1、NICD1、Hes-1的表达受到抑制(P＜0.05).结论:活血散结方能抑制RPE发生EMT,可能是通过抑制Notch信号通路活性实现.

外文标题：Effects of Huoxue Sanjie Formula on TGF-β-induced epithelial mesenchymal transition of retinal pigment epithelial cells based on Notch signaling pathway

外文摘要：Objective:To observe the effects of Huoxue Sanjie Formula on transforming growth factor-β(TGF-β)induced epithelium mesenchymal transition(EMT)of retinal pigment epithelial cells(RPE)through the morphological changes of the cytoskeleton and the expression of cell characteristic proteins,and to clarify the mechanism of Huoxue Sanjie Formula from the expression of related proteins activated by Notch signaling pathway.Methods:Primary RPE were extracted in vitro and subcultured.The cultured RPE were divided into blank control group,rabbit plasma group,TGF-β group,TGF-β+drug containing plasma group,TGF-β+SB431542 group,TGF-β+LY411575 group according to different intervention measures.Using immunofluorescence method to observe the EMT,morphological changes of cytoskeleton,and expression of ZO-1 and α-SMA in RPE induced by TGF-β in the serum containing Huoxue Sanjie Formula;Detection of CBF-1,NICD1,Hes-1 mRNA and protein expression using RT-PCR and Western Blot.Results:Compared with the TGF-β group,the morphological fibrosis of the cytoskeleton of RPE cells in the TGF-β+drug containing plasma group was significantly reduced,the expression of F-actin was significantly reduced(P＜0.01),and the cells were arranged in order;the expression of epithelial cell characteristic protein ZO-1 was increased,and the expression α-SMA was reduced(P＜0.01),and the expression of CBF-1,NICD1,and Hes-1 involved in the Notch signaling pathway was inhibited(P＜0.05).Conclusion:Huoxue Sanjie Formula can inhibit the occurrence of EMT in RPE,which may be achieved by inhibiting the activity of Notch signaling pathway.

外文关键词：

Notch signaling pathwayHuoxue Sanjie FormulaTransforming growth factor-β(TGF-β)Retinal pigment epithelial cells(RPE)Epithelial mesenchymal transition(EMT)

作者：

潘坤、彭俊、梁昊、彭清华

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作者单位：

湖南中医药大学,长沙 410208

中医药防治眼耳鼻喉疾病湖南省重点实验室,长沙 410007

湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,长沙 410007

关键词：

Notch信号通路活血散结方转化生长因子-β 视网膜色素上皮细胞上皮-间充质转化

基金：

国家自然科学基金面上项目湖南省教育厅双一流学科优秀青年项目湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心建设项目

项目编号：

8227458821B0376

出版年：

2024

中华中医药杂志

中华中医药学会

中华中医药杂志

CSTPCD北大核心

影响因子：1.135

ISSN：1673-1727

年,卷(期)：2024.39(9)