首页|CD73对体外心成纤维细胞和心肌细胞增殖与衰老的影响

CD73对体外心成纤维细胞和心肌细胞增殖与衰老的影响

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
目的:探讨 CD73 对心成纤维细胞(CFs)和心肌细胞(CMs)增殖和衰老的影响.方法:体外培养大鼠CFs和CMs,构建慢病毒CD73 过表达载体,转染CFs和CMs.实验分为CD73 过表达组(OE组)和对照组(NC组).RT-PCR法检测转染病毒后各组细胞CD73基因表达水平;衰老相关β-半乳糖苷酶染色检测细胞衰老;MTT法检测细胞增殖;非损伤细胞功能分析仪检测细胞生长曲线和倍增时间;流式细胞术检测细胞周期;RT-PCR法检测增殖相关信号分子mRNA表达水平.结果:慢病毒转染CFs和CMs 72 h后,OE组CD73 mRNA 水平明显高于NC组;NC组CFs和CMs衰老细胞数量多于OE组;OE组CMs转染后3～4 d的增殖能力大于NC组;CFs转染后3～4 d的增殖能力为2组无差异;转染第3天CFs细胞周期变化为2组G1期和S期所占细胞比例无差别,G2期所占比例OE组稍高于NC组;OE组CFs和CMs细胞生长曲线均显示生长指数高于NC组,CMs倍增时间2组差别不显著;CMs转染第3天PI3K和Akt mRNA的表达增高,血管内皮生长因子(VEGF)mRNA的表达变化不明显.结论:CD73对CFs增殖无明显影响,但抑制其衰老;CD73促进CMs的增殖且抑制其衰老;CD73可促进CMs高表达PI3K和Akt,但对 VEGF 表达无明显影响.
Effects of CD73 on the proliferation and senescence of cardiac fibroblasts and cardiomyocytes in vitro
Objective:To investigate the effects of CD73 on the proliferation and senescence of cardiac fibroblasts(CFs)and cardiomyocytes(CMs).Methods:Rats CFs and CMs were cultured in vivo.The lentiviral vector with CD73 overexpression was constructed and transfected into CFs and CMs.Experimental groups were the CD73 overexpression group(OE group)and negative control group(NC group).The gene expression of CD73 in both groups was detected by RT-PCR assay.The cell senescence was detected by senescence-associated β-galactosidase staining.The cell proliferation was detected by MTT assay.The cell growth curve and doubling time were measured by a non-invasive cell function analyzer.The cell cycle was detected by flow cytometry.The RT-PCR method was used to detect the changes in the mRNA expression of proliferation-related signal molecules.Results:Seventy-two hours after the lentiviral transfection of CFs and CMs,the levels of CD73 mRNA in the OE group were significantly higher than those in the NC group.The number of senescent CFs and CMs cells in the NC group was higher than that in the OE group.The proliferative ability of CMs after transfection for 3-4 d was higher in the OE group than that in the NC group,while there was no significant difference between the OE group and the NC group in the proliferative ability of CFs.On the 3rd day after transfection,the cell cycle changes were as follows:There was no significant difference in the proportion of CFs cells in the G1 and S phases of both groups,while the proportion of CFs cells in the G2 phase was slightly higher in the OE group than that in the NC group.The growth curves of CFs and CMs both showed that the OE group had higher growth rates than those in the NC group.The doubling time of CMs in the OE group was shorter than that of the NC group,but the difference in CFs was not significant.On the 3rd day after transfection,the expression of PI3K and Akt mRNA were increased in CMs,while the expression of vascular endothelial growth factor(VEGF)mRNA was unaffected.Conclusion:CD73 has no significant effect on the proliferation of CFs but inhibits their senescence.CD73 promotes CMs proliferation and inhibits their senescence.CD73 can promote the high expression of PI3K and Akt in CMs,but it has no effect on VEGF expression.

CD73proliferationsenescencecardiomyocytescardiac fibroblastsflow cytometrymechanism

王琰、赵育洁、胥良、牛思泉、王地、杨少华、秦续谭、郭志坤

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郑州市第七人民医院,心内科,郑州 450000

新乡医学院河南省医用组织再生重点实验室,新乡 453000

郑州市第七人民医院,河南省心律失常医学重点实验室,郑州 450000

CD73 增殖 衰老 心肌细胞 心成纤维细胞 流式细胞术 机制

国家自然科学基金河南省卫生厅科技攻关计划河南省自然科学基金青年基金河南省医学科技攻关省部共建青年项目河南省科技攻关计划郑州市科技惠民项目郑州市名医支持专项(第二层次)(2021)

8157026815210231011212300410305SBGJ2023030502221023106672022KJHM0033

2024

10.3969/j.issn.1001-1633.2024.01.003

解剖学杂志
中国解剖学会

解剖学杂志

CSTPCD
影响因子：0.407
ISSN：1001-1633
年,卷(期)：2024.47(1)
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