干扰和过表达端粒酶卡哈尔体蛋白1小鼠骨髓间充质干细胞的生物学特性对比  

Comparison of biological characteristics of mouse bone marrow mesenchymal stem cells after interference and overexpression of telomere Cajal body protein-1

作　　者：林书倩 赵玺龙 高景 潘兴华 李自安 阮光萍 Lin Shuqian;Zhao Xilong;Gao Jing;Pan Xinghua;Li Zian;Ruan Guangping(Basic Medical Laboratory of 920 Hospital of Joint Logistics Support Force of Chinese PLA,Kunming 650032,Yunnan Province,China;Integrated Engineering Research Center of Cell Biological Medicine of State and Regions,Kunming 650032,Yunnan Province,China;Transfer Medicine Key Laboratory of Cell Therapy Technology of Yunnan Province,Kunming 650032,Yunnan Province,China)

机构地区：[1]解放军联勤保障部队第九二〇医院基础医学实验室,云南省昆明市650032 [2]干细胞与免疫细胞生物医药技术国家地方联合工程研究中心,云南省昆明市650032 [3]云南省细胞治疗技术转化医学重点实验室,云南省昆明市650032

出　　处：《中国组织工程研究》2025年第31期6616-6624,共9页Chinese Journal of Tissue Engineering Research

基　　金：云南省科技计划项目重大科技专项(2018ZF007),项目负责人:潘兴华;云南省重点项目资助课题(202101AS070039),项目负责人:阮光萍。

摘　　要：背景:随着年龄的增长,骨髓间充质干细胞的功能逐渐降低,延缓骨髓间充质干细胞自身衰老成为一项重要课题。目的:通过改变端粒酶卡哈尔体蛋白1(telomerase cajal body protein 1,TCAB1)基因的表达,探讨延缓骨髓间充质干细胞衰老的途径。方法:细胞贴壁法培养小鼠骨髓间充质干细胞,应用重组慢病毒技术过表达或干扰骨髓间充质干细胞中的TCAB1基因,采用qPCR检测衰老相关基因P16、P21、P53、P27的表达,qPCR检测Telomere的表达判断端粒的相对长度;Western blot检测衰老蛋白P16、P21、P53、P27的表达;CCK-8法检测细胞增殖情况;Annexin V-PE/7-AAD细胞凋亡试剂盒检测细胞凋亡情况。结果与结论:过表达TCAB1基因骨髓间充质干细胞中衰老相关基因及蛋白的表达量下降,端粒相对长度增加,细胞增殖能力较强,细胞凋亡数量较少,细胞呈现年轻状态;干扰TCAB1基因骨髓间充质干细胞中衰老相关基因及蛋白的表达量上升,端粒相对长度缩短,细胞增殖能力较弱,细胞凋亡数量较多,细胞呈现衰老状态。以上结果表明,在适当范围内增加TCAB1的表达量,可以延缓细胞衰老的速度。BACKGROUND:With the increase of age,the function of bone marrow-derived mesenchymal stem cells is gradually reduced,and delaying the aging of bone marrow-derived mesenchymal stem cells itself has become an important topic.OBJECTIVE:To explore ways to delay the aging of bone marrow-derived mesenchymal stem cells by changing the expression of telomerase Cajal body protein 1(TCAB1)gene.METHODS:Mouse bone marrow-derived mesenchymal stem cells were cultured by cell adhesion method.TCAB1 gene in bone marrow-derived mesenchymal stem cells was overexpressed and interfered by recombinant lentivirus technique.The expression of aging related genes P16,P21,P53,and P27 was detected by qPCR.The relative length of telomeres was detected by qPCR.The expression of aging proteins P16,P21,P53,and P27 was detected by western blot assay.Cell proliferation was detected by CCK-8 assay.Annexin V-PE/7-AAD apoptosis kit was used to detect the degree of cell apoptosis.RESULTS AND CONCLUSION:Bone marrow-derived mesenchymal stem cell lines overexpressing TCAB1 gene had decreased expression of senescence related genes and proteins,increased Telomere relative length,stronger cell proliferation,less apoptosis,and a youthful state.The expression of age-related genes and proteins in bone marrow mesenchymal stem cells interfering with TCAB1 gene increased,and the relative telomere length decreased;cell proliferation ability was weak;cell apoptosis was more,and cells showed senescence.These results indicate that increasing the expression of TCAB1 in an appropriate range can delay the rate of cell senescence.

关 键 词：骨髓间充质干细胞 端粒酶卡哈尔体蛋白1 衰老 过表达 干扰 细胞增殖 细胞凋亡 工程化干细胞 

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