抑制Rho/ROCK通路促进酒精干预星形胶质细胞内吞功能及相关基因Vps37c表达  

Inhibition of the Rho/ROCK pathway regulates alcohol intervention in astrocytosis function and expression of the related gene Vps37c

作　　者：戚询中 范景鑫 孙光涛 郭淼 朱晓峰 QI Xun-zhong(The First Affiliated Hospital of Jiamusi University Jiamusi,Jiamusi,154007,China)

机构地区：[1]佳木斯大学附属第一医院 [2]佳木斯大学,黑龙江佳木斯154007 [3]牡丹江医学院,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2023年第6期21-25,共5页Journal of Mudanjiang Medical University

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(2019-KYYWF-1357)。

摘　　要：目的分析酒精及Rho/ROCK路抑制对星形胶质细胞转录组的影响。方法本研究以CTX TNA2大鼠星形胶质细胞为研究对象,选用酒精及Rho/ROCK通路抑制剂法舒地尔进行干预,实验分为对照组、酒精组及法舒地尔组;各组样本干预后Trizol法提取样本总RNA,基于HiSeq平台进行Illumina转录组测序;利用KEGG数据库进行注释及富集分析;qRT-PCR对关键差异基因进行验证。结果酒精组与对照组相比较,获得差异基因14个,KEGG富集分析结果显示,氨基糖和核苷酸糖代谢通路及细胞内吞通路富集程度最高;法舒地尔组与酒精组相比较,获得差异基因149个,KEGG富集分析结果显示,细胞内吞通路排第6位;对照组与酒精组差异基因和酒精组与法舒地尔组差异基因进行差异分析,共获得共有差异基因5个,KEGG富集分析结果显示,细胞内吞通路通路富集程度最高;该通路涉及基因为Vps37c,对该基因进行qRT-PCR验证,结果显示,与对照组相比较,酒精干预后星形胶质细胞Vps37c mRNA表达降低;与酒精组相比较,法舒地尔干预后Vps37c mRNA表达升高。此结果与测序结果一致。结论酒精干预能够抑制星形胶质细胞内吞功能,抑制细胞内吞功能基因Vps37c表达;Rho/ROCK通路抑制能够促进星形胶质细胞内吞功能,促进细胞内吞功能基因Vps37c表达。Objective To analyze the effect of alcohol and Rho/ROCK pathway inhibitors on the transcriptome of astrocytes.Methods CTX TNA2 rat astrocytes were treated with alcohol and Fasudil,a Rho/ROCK pathway inhibitor.The experiment was divided into control group,alcohol group and Fasudil group.After the intervention,total RNA was extracted using the Trizol method,and Illumina sequencing was performed based on the HiSeq platform.The KEGG database was used for annotation and enrichment analysis.Key differential genes were validated by qRT-PCR.Results Fourteen different genes were identified in the alcohol group compared to the control group.KEGG enrichment analysis showed that the enrichment of Amino sugar and nucleotide sugar metabolism pathway and Endocytosis pathway were the highest.Compared with alcohol group,149 differential genes were obtained in Fasudil group.The KEGG enrichment analysis showed that Endocytosis ranked sixth.The differential expression genes between the control group and the alcohol group and between the alcohol group and the Fasudil group were compared,and a total of 5 differential genes were obtained.KEGG enrichment analysis revealed the highest enrichment in the Endocytosis pathway.The gene involved in this pathway is Vps37c,which was validated by qRT-PCR.The results showed a reduction in mRNA expression of Vps37c in astrocytes after alcohol intervention compared to the control group.The expression of Vps37c mRNA increased after Fasudil intervention compared to the alcohol group.The results are in agreement with the sequencing results.Conclusion Alcohol intervention modulates astrocyte endocytosis function and inhibits Vps37c expression.Rho/ROCK pathway inhibitors can regulate endocytosis in astrocytes and promote Vps37c expression.

关 键 词：酒精 星形胶质细胞 Rho/ROCK通路 细胞内吞 Vps37c 

分 类 号：R749.1[医药卫生—神经病学与精神病学]