摘要
目的观察急性肺损伤(acute lung injury,ALI)时miRNA-21通过磷酸酯酶一张力蛋白同源物(phosphatase and tensin homolog,PTEN)调控PI13K/AKT信号通路途径影响间充质干细胞(mesenchymal stem cells,MSC)存活情况。方法选择体外通过H2O2/SD刺激MSC,观察MSC凋亡和增殖情况,然后通过转染上调和下调MSC的miR-21表达,观察miR-21靶基因PTEN及信号通路分子PI3K/Akt、Bcl-2表达的改变,以及对MSC凋亡和增殖的影响,明确miR-21对ALI时MSC凋亡和增殖的调节及其作用机制;将转染后高表达miR-21的MSC注入LPS诱导的ALI小鼠体内,观察转染后的MSC在体内凋亡及增殖,miR-21靶基因及相关信号通路分子表达的改变,及修复肺损伤和保护肺功能的作用。结果 ALI组、MSC处理组的PTEN、PI3K、Akt、Bcl2基因和蛋白表达高于空白对照组,差异具有统计学意义(P <0.05);MSC处理组的PTEN、PI3K、Akt、Bcl2基因表达低于ALI组,差异具有统计学意义(P <0.05);LY294002处理的miR-21-MSC标记细胞、沉默PTEN的miR-21-MSC标记细胞的MSC存留率、MSC移植后细胞存活率高于miR-21-MSC、野生型MSC细胞,肺湿/干重比低于miR-21-MSC、野生型MSC细胞,差异具有统计学意义(P <0.05)。结论探讨ALI时调节MSC凋亡、增殖相关的miRNAs及其作用机制有助于为增加移植后的细胞存活率找到新的治疗靶点。
Objective To observe the effect of miRNA-21 on the survival of mesenchymal stem cells(MSC) after acute lung injury(ALI) by regulating PI13 K/AKT signaling pathway by phosphatase and tensin homolog(PTEN). Methods MSCs were stimulated with H2 O2/SD in vitro to observe the apoptosis and proliferation of MSCs. Then, the expression of miR-21 was up-regulated and down regulated by transfection.The expression of PTEN, PI13 K/AKT and Bcl-2, as well as the effect on apoptosis and proliferation of MSCs were observed.The regulation and mechanism of miR-21 on apoptosis and proliferation of MSCs in Ali were clarified After transfection into LPS induced Ali mice,the apoptosis and proliferation of MSC, the expression of miR-21 target gene and related signaling pathway molecules,the repair of lung injury and the protection of lung function were observed. Results The gene and protein expressions of PTEN, PI3 K, Akt and Bc12 in ALI group and MSC group were significantly higher than those in blank control group(P < 0.05);the gene expressions of PTEN, PI3 K, Akt and Bc12 in MSC group were significantly lower than those in ALI group(P < 0.05);the survival rate and survival rate of miR-21-MSC labeled cells and PTEN silenced miR-21-MSC labeled cells treated with LY294002 were significantly higher than those of miR-21-MSC and wild-type MSC, and the wet/dry weight ratio of lung was significantly lower than that of miR-21-MSC and wild-type MSC(P < 0.05). Conclusion To explore the miRNAs related to apoptosis and proliferation of MSC during ALI and its mechanism will help to find a new therapeutic target for increasing the survival rate of transplanted cells.
作者
邵建平
蔡施霞
孙运波
方巍
刘冰
李连弟
SHAO Jianping;CAI Shixia;SUN Yunbo;FANG Wei;LIU Bing;LI Liandi(Department of Critical Care Medicine,Affiliated Hospital of Qingdao University,Qingdao Shandong 266003,China)
出处
《中国卫生标准管理》
2021年第14期125-129,共5页
China Health Standard Management
基金
国家自然科学基金青年科学基金项目(81701879)。
