首页|间充质干细胞外泌体通过微小RNA-125b-5p抑制类风湿关节炎成纤维样滑膜细胞的增殖、侵袭和炎症

间充质干细胞外泌体通过微小RNA-125b-5p抑制类风湿关节炎成纤维样滑膜细胞的增殖、侵袭和炎症

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目的 探讨间充质干细胞来源外泌体(MSC-EVs)对类风湿关节炎(RA)成纤维样滑膜细胞MH7A的生物学作用及其机制.方法 将MH7A细胞系分为对照组和外泌体组.对MSC的培养上清液采用差速离心法收集MSC-EVs,对照组采用常规方法进行培养,外泌体组则将MH7A细胞与MSC-EVs共同孵育24 h,2组细胞达到收样条件后通过免疫荧光实验、Transwell实验、酶联免疫吸附实验(ELISA)及实时荧光定量聚合酶链反应(qRT-PCR)等实验观察MSC-EVs对MH7A细胞株生物学行为的影响及其分子机制,各观察指标采用独立样本t检验进行统计学分析.结果 粒径分析检测及透射电镜结果表明,通过差速离心法成功提取了 MSC-EVs,随后免疫荧光证实MSC-EVs能被MH7A细胞株所摄取.细胞增殖结果表明,MSC-EVs组细胞核增殖抗原(Ki-67)荧光灰度值(4.01±1.01)明显低于对照组(11.67±1.53),差异有统计学意义(t=-7.273,P<0.01).Transwell结果显示,MSC-EVs组侵袭能力(70.00±5.29)显著低于对照组(110.67±6.11),差异有统计学意义(t=-8.714,P<0.01).ELISA结果表明,MSC-EVs组的炎性因子低于对照组,差异有统计学意义[肿瘤坏死因子-α(TNF-α):t=-5.345,P<0.01;白细胞介素(IL)-1β:t=-3.742,P<0.05;IL-6:t=-7.579,P<0.01;IL-8:t=-6.390,P<0.01].qRT-PCR 结果表明,MSC-EVs 组MH7A细胞内微小RNA(miR)-125b-5p的表达水平(14.83±2.48)高于对照组(1.01±0.15),差异有统计学意义(t=-8.714,P<0.01).结论 MSC-EVs能通过上调miR-125b-5p的表达进而抑制MH7A细胞的增殖、侵袭和炎症.
Exosomes derived from mesenchymal stem cells inhibit the proliferation,invasion and inflammation of rheumatoid arthritis fibroblast-like synoviocytes through microRNA-125b-5p
Objective To investigate the biological effects and mechanisms of mesenchymal stem cell-extracellular vesicles(MSC-EVs)on rheumatoid arthritis(RA)fibroblast-like synoviocytes(MH7A).Methods MH7A cell line was divided into control group and MSC-EVs group.MSC-EVs were collected from the culture supernatant of MSC by differential centrifugation.The control group was cultured with rou-tine methods,and MH7A cells were incubated with MSC-EVs for 24 h in the MSC-EVs group.After the two groups of cells reached the sample collection conditions,the immunofluorescence experiment,Tran-swell experiment,enzyme-linked immunosorbent assay(ELISA),and quantitative real-time polymerase chain reaction(qRT-PCR)were used to observe the effect of MSC-EVs on the biological behavior of MH7A cells and its molecular mechanism.The independent sample t test was used for statistical analysis.Results The results of nanoparticle tracking analysis and transmission electron microscopy showed that MSC-EVs were successfully extracted by differential centrifugation,and then immunofluorescence confirmed that MSC-EVs could be uptaken by MH7A cells.The results of cell proliferation showed that the gray value of proliferation cell nuclear antigen(Ki-67)fluorescence in the MSC-EVs group(4.01±1.01)was signifi-cantly lower than that in the control group(11.67±1.53)(4=-7.273,P<0.01).Transwell results showed that the invasion ability of MSC-EVs group(70.00±5.29)was significantly lower than that of the control group(110.67±6.11)(t=-8.714,P<0.01).ELISA results showed that the levels of inflam-matory factors in the MSC-EVs group were significantly lower than those in the control group[tumor necro-sis factor-α(TNF-α):t=-5.345,P<0.01;interleukin(IL)-1β:t=-3.742,P<0.05;IL-6:t=-7.579,P<0.01;IL-8:t=-6.390,P<0.01].The results of qRT-PCR showed that the expression level of microRNA(miR)-125b-5p in the MSC-EVs group(14.83±2.48)was significantly higher than that in the control group(1.01±0.15)(t=-8.714,P<0.01).Conclusion MSC-EVs could inhibit the prolifera-tion,invasion and inflammation of MH7A cells by up-regulating the expression of miR-125b-5p.

Rheumatoid arthritisMesenchymal stem cellsExosomesInflammatory factors

包德明、宋瑞鹏、刘鸣、宋辉、赵世新、王丹

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郑州大学第一附属医院骨科,郑州 450052

类风湿关节炎 间充质干细胞 外泌体 炎性因子

2024

中华实验外科杂志
中华医学会

中华实验外科杂志

CSTPCD
影响因子:0.759
ISSN:1001-9030
年,卷(期):2024.41(2)
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