年轻大鼠骨髓间充质干细胞来源外泌体逆转老龄大鼠骨髓间充质干细胞衰老  

Exosomes derived from bone marrow mesenchymal stem cells of young rats to reverse senescence in aged rat bone marrow mesenchymal stem cells

作　　者：张熊劲夫 陈奕达 程歆怡 刘岱珲 施勤[1] Zhang Xiongjinfu;Chen Yida;Cheng Xinyi;Liu Daihui;Shi Qin(Department of Orthopedics,First Affiliated Hospital of Soochow University,Institute of Orthopedics of Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,苏州大学骨科研究所,江苏省苏州市215006

出　　处：《中国组织工程研究》2025年第36期7709-7718,共10页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82172485),项目负责人:施勤。

摘　　要：背景:骨髓间充质干细胞是骨形成的主要效应细胞,随着年龄的增加,骨髓间充质干细胞的再生能力减弱、分化功能受损,导致骨质疏松。因此,恢复老龄骨髓间充质干细胞的再生能力和细胞功能对骨质疏松的有效治疗至关重要。目的:探究年轻大鼠第3,11代骨髓间充质干细胞来源外泌体对老年大鼠骨髓间充质干细胞衰老的影响。方法:分离培养6-8周龄雌性SD大鼠骨髓间充质干细胞,分别传代至第3代和第11代,随后提取第3,11代骨髓间充质干细胞来源外泌体。分离培养18月龄雌性SD大鼠骨髓间充质干细胞,传代至第3代,分为3组:对照组为常规培养,其他2组用第3,11代骨髓间充质干细胞来源外泌体干预。外泌体干预48 h后,利用β-半乳糖苷酶染色试剂盒检测细胞核内β-半乳糖苷酶的表达,qRT-PCR检测衰老相关基因的表达。通过Small RNA测序,比较第3,11代骨髓间充质干细胞来源外泌体内miRNA的表达差异。结果与结论:(1)与对照组和第11代骨髓间充质干细胞来源外泌体组相比,第3代骨髓间充质干细胞来源外泌体组老龄大鼠骨髓间充质干细胞的β-半乳糖苷酶活性显著降低;(2)与对照组相比,第3代骨髓间充质干细胞来源外泌体组衰老相关基因p21和p16的表达显著降低(P<0.05),而第11代骨髓间充质干细胞来源外泌体组衰老相关基因p21和p16的表达无显著差异;(3)测序结果表明,两种外泌体内miRNA的表达存在显著差异,其中表达差异最显著的miRNA是let-7c-5p、let-7b-5p 、miR-320-3p和miR-26a-5p,KEGG分析结果显示显著性差异miRNA富集通路包含mTOR、AMPK等衰老相关信号通路。上述结果表明,第3代骨髓间充质干细胞来源外泌体具有逆转老龄大鼠骨髓间充质干细胞衰老的能力。BACKGROUND:Bone marrow mesenchymal stem cells are the main effector cells for bone formation.With the increase of age,the regenerative ability of bone marrow mesenchymal stem cells is weakened and the differentiation function is impaired,leading to poor osteoporosis.Therefore,restoring the regenerative capacity and cellular function of aged bone marrow mesenchymal stem cells is essential for the effective treatment of osteoporosis.OBJECTIVE:To investigate the effects of passage 3 and passage 11 bone marrow mesenchymal stem cells-derived exosomes of young rats on the aging of bone marrow mesenchymal stem cells derived from elderly rats.METHODS:Bone marrow mesenchymal stem cells from 6-8-week-old female SD rats were isolated and cultured,and passaged to the passages 3 and 11,respectively.Then,exosomes from passages 3 and 11 bone marrow mesenchymal stem cells were extracted.Bone marrow mesenchymal stem cells from 18-month-old female SD rats were isolated and cultured,passaged to passage 3,and divided into 3 groups.The control group was routinely cultured,and the other two groups were intervened with exosomes from passages 3 and 11 bone marrow mesenchymal stem cells.After 48 hours of exosome intervention,the expression ofβ-galactosidase in the nucleus was detected byβ-galactosidase staining kit.The expression of aging-related genes was detected by qRT-PCR.The expression differences of miRNA in exosomes from passages 3 and 11 bone marrow mesenchymal stem cells were compared by Small RNA sequencing.RESULTS AND CONCLUSION:(1)Compared with the control group and passage 11 bone marrow mesenchymal stem cell-derived exosomes group,theβ-galactosidase activity of bone marrow mesenchymal stem cells of aged rats was significantly lower in the passage 3 bone marrow mesenchymal stem cell-derived exosomes group.(2)Compared with the control group,the expression of aging-related genes p21 and p16 was significantly reduced in the passage 3 bone marrow mesenchymal stem cell-derived exosome group(P<0.05),while there was no significa

关 键 词：骨质疏松 外泌体 骨髓间充质干细胞 miRNA 衰老 工程化细胞外囊泡 

分 类 号：R459.9[医药卫生—治疗学] R310.17[医药卫生—临床医学] R180.21