Page 10 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第2期
·160 · Journal of Nanjing Medical University（Natural Sciences） 2021年2月

·基础研究·

GCN5调控sublytic C5b⁃9诱导大鼠肾小球系膜细胞表达Cyclin
D1及致SOX9的乙酰化修饰

郭苗苗，刘龙飞，谢梦晓，吴志皎，罗 灿，彭明玉，赵 聃，张 婧，邱 文，王迎伟 *
南京医科大学免疫学系，江苏 南京 211166

［摘 要］ 目的：探讨GCN5调控sublytic C5b⁃9刺激大鼠肾小球系膜细胞（glomerular mesangial cell，GMC）诱导Cyclin D1基因
表达以及GCN5乙酰化修饰SOX9转录因子的作用。方法：先用Western blot检测Thy⁃1肾炎（Thy⁃1 nephritis，Thy⁃1N）大鼠的肾
组织和sublytic C5b⁃9 刺激的GMC中GCN5、SOX9和Cyclin D1的表达。然后分别将pIRES2⁃GCN5过表达和shGCN5小干扰质
粒或将这些质粒与Cyclin D1启动子质粒行不同组合转染GMC，用荧光素酶报告基因实验和real⁃time PCR、Western blot检测过
表达或沉默GCN5后对Cyclin D1表达的影响。同时用免疫共沉淀（Co⁃IP）和Western blot 检测sublytic C5b⁃9刺激或分别转染
GCN5 过表达和酶活性突变质粒（ΔGCN5）的 GMC 中 SOX9 的乙酰化修饰。结果：Thy⁃1N 大鼠肾组织和 sublytic C5b⁃9 刺激的
GMC中GCN5、SOX9、Cyclin D1蛋白水平均明显上调，而过表达GCN5基因能上调Cyclin D1的启动子活性、mRNA和蛋白表达；
敲低GCN5表达则得到了相反的结果。此外，sublytic C5b⁃9刺激和过表达GCN5基因促进了SOX9的乙酰化修饰，而沉默GCN5后
则降低了SOX9的乙酰化水平。结论：GCN5可上调GMC中Cyclin D1的表达，并促进SOX9的乙酰化修饰。
［关键词］ GCN5；SOX 9；sublytic C5b⁃9；Cyclin D1；乙酰化修饰
［中图分类号］ R392.11 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）02⁃160⁃06
doi：10.7655/NYDXBNS20210202

Cyclin D1 expression and SOX9 acetylation regulated by GCN5 in glomerular mesangial

cell induced by sublytic C5b⁃9
GUO Miaomiao，LIU Longfei，XIE Mengxiao，WU Zhijiao，LUO Can，PENG Mingyu，ZHAO Dan，ZHANG Jing，
QIU Wen，WANG Yingwei *
Department of Immunology，Nanjing Medical University，Nanjing 211166，China

［Abstract］ Objective：This study aims to explore the effects of Cyclin D1 expression regulated by GCN5 and the acetylation
modification of SOX9 by GCN5 in the glomerular mesangial cells（GMC）stimulated by sublytic C5b⁃9. Methods：Firstly，GCN5，SOX9
and Cyclin D1 expression in the renal tissue of Thy⁃1 nephritis（Thy⁃1N）rats（in vivo）and in sublytic C5b⁃9⁃treated GMC（in vitro）was
examined using Western blot. Then the plasmids of pIRES2⁃GCN5 and shGCN5 or these plasmids with Cyclin D1 promoter plasmids
were respectively transfected rat GMC，and Cyclin D1 gene initiation，transcription and expression were measured by luciferase
reporter gene assay，real⁃time PCR and Western blot. Meanwhile，SOX9 acetylation was detected by co⁃immunoprecipitation（Co⁃IP）
and Western blot in the GMC exposed to sublytic C5b⁃9 or transfected GCN5 overexpression or enzyme activity mutation（ΔGCN5）
plasmids. Results：The expression level of GCN5，SOX9 and Cyclin D1 was significantly up⁃regulated both in vivo and in vitro，and
GCN5 gene overexpression increased Cyclin D1 promoter activity，mRNA transcription and protein expression；GCN5 gene knockdown
made for the opposite results. Besides，SOX9 acetylation was also elevated upon sublytic C5b ⁃ 9 stimulation and GCN5 gene
overexpression；SOX9 acetylation was reduced upon GCN5 knockdown. Conclusion：GCN5 can promote Cyclin D1 expression and
acetylate SOX9 in GMC.
［Key words］ GCN5；SOX9；sublytic C5b⁃9；Cyclin D1；acetylation
［J Nanjing Med Univ，2021，41（02）：160⁃165］

［基金项目］ 国家自然科学基金（31770934，81971468）
∗
通信作者（Corresponding author），E⁃mail：wangyw1508@njmu.edu.cn