基于转录组学探讨柴姜方调控TGF-β/Rho-ROCK通路对青光眼滤过术后滤过泡瘢痕化的抑制作用  

Inhibitory Effect of Chaijiang Prescription on Scarring of Filtering Blebs after Glau⁃coma Filtration Surgery by Regulating TGF-β/Rho-ROCK Pathway Based on Transcriptomic Analysis

作　　者：张伟道 李欣[1] 陈杰[1] 冯俊[1] ZHANG Wei-dao;LI Xin;CHEN Jie;FENG Jun(Eye Hospital of China Academy of Chinese Medical Sciences,Beijing 100040)

机构地区：[1]中国中医科学院眼科医院,北京100040

出　　处：《世界中西医结合杂志》2025年第1期37-45,共9页World Journal of Integrated Traditional and Western Medicine

基　　金：国家中医药管理局眼体同调同治重点研究室;中国中医科学院科技创新工程重大攻关项目(CI2021A02613);中国中医科学院眼科医院院内课题(1011632)。

摘　　要：目的探讨柴姜方基于转录组学调控TGF-β/Rho-ROCK通路对青光眼滤过术后抗瘢痕化的分子机制。方法将57只SPF级远交系健康、无眼疾的成年新西兰长耳白兔,按随机数字表法分为空白组9只,模型组、柴姜方低剂量组、柴姜方中剂量组、柴姜方高剂量组各12只。青光眼滤过手术造模成功后,于术前和术后1、3、7、14 d测量术眼眼压,并利用裂隙灯显微镜观察手术滤过泡形成情况。于术后14 d处死兔子并获取手术区标本组织,通过HE染色和Masson染色评价滤过通道组织病理结构变化;转录组学筛选手术区标本组织差异表达基因并富集差异表达通路,运用qRT-PCR、Western blot检测TGF-β1、RhOA、ROCK2、ɑ-SMA mRNA和蛋白的相对表达量。结果眼压测量结果显示,术后7、14 d与模型组比较,柴姜方中、高剂量组眼压均明显降低,差异有统计学意义(P<0.01)。HE染色和Masson染色结果显示,术后14 d与模型组比较,柴姜方中、高剂量组成纤维细胞数目和胶原纤维面积占比明显降低,差异有统计学意义(P<0.01)。转录组学结果显示,模型组相对于空白组明显上调基因数448,下调基因数314;柴姜方中剂量组相对于模型组明显上调基因数448,下调基因数792;对各组差异表达基因进行KEGG富集分析发现主要集中于TGF-β/Rho-ROCK通路。qRT-PCR、Western blot检测结果显示,与模型组比较,柴姜方中、高剂量组TGF-β1、RhOA、ROCK2、ɑ-SMA mRNA和蛋白的相对表达量均明显降低,差异有统计学意义(P<0.01)。结论柴姜方中、高剂量组通过抑制TGF-β/Rho-ROCK通路的活化、成纤维细胞和胶原纤维的增殖对青光眼滤过术后发挥抗瘢痕化作用,可有效降低眼压并维持滤过泡功能。Objective To explore the molecular mechanisms of Chaijiang Prescription in resisting scarring by reg⁃ulating the TGF-β/Rho-ROCK pathway after glaucoma filtration surgery based on transcriptomics.Methods Fifty-seven SPF-grade,outbred healthy,eye disease-free adult New Zealand long-eared white rabbits were randomly divided into a blank group(n=9),a model group,and Chaijiang Prescription low-dose,medium-dose,and high-dose groups(n=12 in each group)according to a random number table.Glaucoma filtration surgery was performed to establish the model.In⁃traocular pressure(IOP)was measured preoperatively and on days 1,3,7,and 14 after surgery.The formation of the filtering blebs was observed using a slit-lamp microscope.On day 14 after surgery,the rabbits were euthanized and tissue speci⁃mens from the surgical area were collected.HE and Masson staining were used to evaluate pathological changes in the fil⁃tration channel tissue.Transcriptomic analysis was conducted to identify differentially expressed genes(DEGs)in the surgi⁃cal area tissues,and enrichment of differentially expressed pathways was performed.The relative expression levels of TGF-β1,RhOA,ROCK2,andα-SMA mRNA and proteins were detected using qRT-PCR and Western blot.Results The IOP measurements showed that on days 7 and 14 after surgery,the IOP in the Chaijiang Prescription medium-and high-dose groups was significantly lower than in the model group(P<0.01).HE and Masson staining results indicated that,compared to the model group,the number of fibroblasts and the collagen fiber area in the Chaijiang Prescription medium-and high-dose groups were significantly reduced on day 14 after surgery(P<0.01).Transcriptomic analysis revealed that 448 genes were upregulated and 314 genes were downregulated in the model group compared to the blank group.In the Chaijiang Pre⁃scription medium-dose group,448 genes were upregulated and 792 genes were downregulated compared to the model group.KEGG pathway enrichment analysis showed that the DEGs were mainly concent

关 键 词：青光眼 青光眼滤过手术 柴姜方 TGF-β/Rho-ROCK通路 抗瘢痕化 

分 类 号：R285.5[医药卫生—中药学]