Pax6基因表达对过氧化氢诱导骨髓间充质干细胞衰老的影响  

Effect of Pax6 gene expression on hydrogen peroxide-induced aging in bone marrow mesenchymal stem cells

作　　者：高杰 邹星星[1] 文邦红 李远迪 苏敏 胡蓉 Gao Jie;Zou Xingxing;Wen Banghong;Li Yuandi;Su Min;Hu Rong(Department of Histology and Embryology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;Translational Medicine Research Center,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities,Guiyang 550025,Guizhou Province,China;National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique,Guizhou Province Key Laboratory of Regenerative Medicine,Guizhou Medical University,Key Laboratory of Adult Stem Cell Transformation of Chinese Academy of Medical Sciences,Guiyang 550025,Guizhou Province,China)

机构地区：[1]贵州医科大学基础医学院组织学与胚胎学教研室,贵州省贵阳市550025 [2]贵州医科大学转化医学研究中心,贵州省贵阳市550025 [3]贵州省高等学校自身免疫病研究重点实验室,贵州省贵阳市550025 [4]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,贵州省再生医学重点实验室,中国医学科学院成体干细胞转化研究重点实验室,贵州省贵阳市550025

出　　处：《中国组织工程研究》2024年第31期4921-4925,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(82171343,82060151),项目负责人:苏敏;贵州省科技厅项目(黔科合支撑[2020]4Y230号),项目负责人:苏敏;贵州省科技厅项目(黔科合基础-ZK[2022]一般344),项目负责人:高杰;贵州省高等学校重点实验室(黔教技[2023]016号),项目负责人:苏敏。

摘　　要：背景:多种眼科疾病的发生发展与过度氧化应激密切相关,抑制氧化应激反应可能会产生预防和治疗效果。目的:探究Pax6基因表达对过氧化氢诱导的小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BM-MSCs)衰老的影响。方法:复苏BM-MSCs、Pax6/BM-MSCs、shPax6/BM-MSCs,经过氧化氢处理24 h后进行β-半乳糖苷酶染色,流式细胞术检测增殖指标Ki67的表达和细胞凋亡情况,RT-PCR检测衰老相关分子Wnt7a、p21、p53的表达。结果与结论:①经过氧化氢处理后,3组细胞显示衰老表型,β-半乳糖苷酶染色阳性;与BM-MSCs组相比,Pax6/BM-MSCs组内阳性细胞表达较少,shPax6/BM-MSCs组内阳性细胞表达较多,差异有显著性意义(P<0.05);②流式细胞术结果显示:与BM-MSCs组相比,Pax6/BM-MSCs组细胞内Ki67阳性表达增加,细胞凋亡水平降低,shPax6/BM-MSCs组细胞内Ki67阳性表达降低,细胞凋亡水平升高,差异有显著性意义(P<0.05);③RT-PCR检测结果显示:与BM-MSCs组相比,Pax6/BM-MSCs组的Wnt7a、p53、p21表达量降低,shPax6/BM-MSCs组的Wnt7a、p53、p21表达量增加,差异有显著性意义(P<0.05);④结果表明:Pax6过表达能够对抗过氧化氢诱导的BM-MSCs衰老进程,可能与Wnt信号通路有关。BACKGROUND:The occurrence and development of various ophthalmic diseases are closely related to excessive oxidative stress,and the inhibition of oxidative stress response may produce preventive and therapeutic effects.OBJECTIVE:To explore the role of Pax6 gene expression on hydrogen peroxide-induced aging of mouse bone marrow mesenchymal stem cells(BM-MSCs).METHODS:Resuscitated BM-MSCs,Pax6/BM-MSCs,and shPax6/BM-MSCs were treated with hydrogen peroxide for 24 hours,and thenβ-galactosidase staining was performed.The proliferation index Ki67 expression and apoptosis were detected by flow cytometry.The expression of senescence-associated molecules(Wnt7a,p21,and p53)was detected by RT-PCR.RESULTS AND CONCLUSION:(1)After hydrogen peroxide treatment,the cells of the three groups showed senescence phenotype andβ-galactosidase staining was positive.Compared with BM-MSCs group,the expression of positive cells in Pax6/BM-MSCs group was less and that in the shPax6/BM-MSCs group was more,and the difference was statistically significant(P<0.05).(2)The results of flow cytometry showed that compared with BM-MSCs group,the positive expression of Ki67 in the Pax6/BM-MSCs group increased and the level of apoptosis decreased,while the positive expression of Ki67 decreased and the level of apoptosis increased in the shPax6/BM-MSCs group;the difference was significantly different(P<0.05).(3)RT-PCR showed that compared with the BM-MSCs group,the expression of Wnt7a,p53,and p21 decreased in the Pax6/BM-MSCs group,while the expression of Wnt7a,p53,and p21 increased in the shPax6/BM-MSCs group;the difference was significantly different(P<0.05).(4)These findings indicate that overexpression of Pax6 can antagonize the aging progression of BM-MSCs induced by hydrogen peroxide,which may be related to Wnt signaling pathway.

关 键 词：骨髓间充质干细胞 PAX6 Wnt7a 过氧化氢 凋亡 衰老 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R329.2