摘要
目的研究microRNA-129-5p(miR-129-5p)对脂多糖(LPS)诱导的小鼠肺上皮细胞凋亡和自噬的影响。方法取小鼠肺上皮细胞MLE-12,分别转染miR-129-5p模拟物(设为miR-129-5p mi组)和阴性对照寡核苷酸(设为miR-NC组),随后用500 ng·mL^(-1)LPS处理MLE-12细胞。用细胞计数法-8(CCK-8)检测各组细胞的增殖能力;用流式细胞术检测各组细胞的凋亡率;用蛋白质印迹(Western blot)法检测各组细胞中自噬相关蛋白P62和Bec-lin-1的表达水平。结果miR-NC组和miR-129-5p mi组细胞在24 h的细胞存活率分别为(52.73±1.41)%,(68.70±3.55)%,在48 h的细胞存活率分别为(48.75±7.22)%,(69.24±3.05)%,在72 h的细胞存活率分别为(48.22±4.28)%,(65.61±2.53)%;细胞凋亡率分别为(28.46±2.51)%,(17.24±2.06)%;P62蛋白的表达水平分别为1.01±0.03和1.54±0.07,Beclin-1蛋白的表达水平分别为0.98±0.09和0.52±0.05,差异均有统计学意义(均P<0.05)。结论过表达miR-129-5p可显著促进LPS诱导小鼠肺上皮细胞MLE-12增殖,并抑制细胞凋亡和自噬,从而改善LPS诱导的急性肺损伤。
Objective To investigate the impact of microRNA-129-5 p(miR-129-5 p) on cell apoptosis and autophagy caused by lipopolysaccharide(LPS) in MLE-12 cells. Methods Mouse lung epithelial cells MLE-12 cells were transfected with miR-129-5 p mimics(miR-129-5 p mi group) and its negative control(miR-NC group), then cells were treated with LPS(500 ng·mL;). Cell counting kit-8(CCK-8) method was used to detect the proliferation of each group of cells, flow cytometry was conducted to detect the apoptosis rate of cells in each group, Western blot was utilized to detect the expression of autophagy-associated proteins P62 and Beclin-1 in each group of cells. Results The cell survival rates at 24 h in miR-NC group and miR-129-5 p mi group were(52.73±1.41)% and(68.70±3.55)%, respectively;the cell survival rates at 48 h were(48.75±7.22)% and(69.24±3.05)%, respectively;the cell survival rates at 72 h were(48.22±4.28)% and(65.61±2.53)%;the apoptosis rates were(28.46±2.51)% and(17.24±2.06)%;the expression levels of P62 protein were 1. 01 ± 0. 03 and 1. 54 ± 0. 07,the expression levels of Beclin-1 protein were0. 98 ± 0. 09 and 0. 52 ± 0. 05,the differences were all statistically significant( all P < 0. 05). Conclusion Transfection of miR-129-5 p mimics can significantly promote the proliferation of mouse lung epithelial cells MLE-12,and inhibit cell apoptosis and autophagy,thereby improving the acute lung injury induced by LPS.
作者
张晓强
余建平
ZHANG Xiao-qiang;YU Jian-ping(Department of Infectious Diseases,Yuhang District,The Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 311100,Zhejiang Province,China;Department of Infectious Diseases,The First People’s Hospital of Yuhang District,Hangzhou 311100,Zhejiang Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第20期2752-2754,共3页
The Chinese Journal of Clinical Pharmacology
