miR-29a-3p靶向调控FSTL1对肺炎链球菌诱导的肺泡上皮细胞损伤的影响OA
Effect of miR-29a-3p on Streptococcus pneumonia induced alveolar epithelial cell injury by target regulating FSTL1
目的:探讨miR-29a-3p调节卵泡蛋白样蛋白1(FSTL1)对肺炎链球菌(SP)诱导的肺泡上皮细胞损伤的影响.方法:将HPAEpiC细胞分为对照(Ctrl)组、SP组、SP+miR-NC组、SP+miR-29a-3p mimic组、SP+miR-29a-3p mimic+OE-NC组、SP+miR-29a-3p mimic+FSTL1-OE组.qRT-PCR检测细胞中miR-29a-3p和FSTL1 mRNA水平;CCK-8法测定各组细胞增殖能力;Annexin Ⅴ-PI法检测各组细胞凋亡水平;ELISA检测各组细胞中TNF-α和IL-6水平;试剂盒检测各组细胞中SOD活性及ROS生成量;Western blot检测各组细胞中FSTL1、Bax和Bcl-2蛋白表达水平;双荧光素酶实验测定miR-29a-3p与FSTL1的靶向关系.结果:与Ctrl组相比,SP、SP+miR-NC、SP+miR-29a-3p mimic、SP+miR-29a-3p mimic+OE-NC、SP+miR-29a-3p mimic+FSTL1-OE组细胞中增殖抑制率、细胞凋亡率、TNF-α、IL-6、ROS、Bax和FSTL1 mRNA及蛋白表达水平显著升高(P<0.05),miR-29a-3p水平、细胞存活率、SOD和Bcl-2水平显著降低(P<0.05);与SP+miR-NC组相比,SP+miR-29a-3p mimic组细胞中增殖抑制率、细胞凋亡率、TNF-α、IL-6、ROS、Bax和FSTL1 mRNA及蛋白表达水平显著降低(P<0.05),miR-29a-3p水平、细胞存活率、SOD和Bcl-2水平显著升高(P<0.05);与SP+miR-29a-3p mimic+OE-NC组相比,SP+miR-29a-3p mimic+FSTL1-OE组细胞中增殖抑制率、细胞凋亡率、TNF-α、IL-6、ROS、Bax和FSTL1 mRNA及蛋白表达水平显著升高(P<0.05),miR-29a-3p水平、细胞存活率、SOD和Bcl-2水平显著降低(P<0.05);与FSTL1-WT和miR-NC共转染细胞相比,FSTL1-WT和miR-29a-3p mimic共转染组细胞中荧光酶活性显著降低(P<0.05).结论:miR-29a-3p可能通过靶向下调FSTL1减轻SP诱导的肺泡上皮细胞损伤,促进细胞增殖,抑制细胞凋亡和炎症反应.
Objective:To investigate the effect of miR-29a-3p on alveolar epithelial cell injury induced by Streptococcus pneumonia(SP)by regulating follistatin-like protein 1(FSTL1).Methods:HPAEpiC cells were divided into control(Ctrl)group,SP group,SP+miR-NC group,SP+miR-29a-3p mimic group,SP+miR-29a-3p mimic+OE-NC group,and SP+miR-29a-3p mimic+FSTL1-OE group.qRT-PCR was used to detect levels of miR-29a-3p and FSTL1 mRNA in cells.CCK-8 methods was used to determine prolif-eration ability of cells in each group.Annexin Ⅴ-PI method was used to detect level of apoptosis of cells in each group.ELISA was used to detect levels of TNF-α and IL-6 of cells in each group.SOD activity and the amount of ROS of cells in each group were detected by kits.Western blot was used to detect expression levels of FSTL1,Bax and Bcl-2 proteins of cells in each group.Dual luciferase assay was used to determine the targeting relationship between miR-29a-3p and FSTL1.Results:Compared with Ctrl group,proliferation inhibition rate,apoptosis rate,TNF-α,IL-6,ROS,Bax and FSTL1 mRNA and protein expression levels were significantly higher in SP group,SP+miR-NC group,SP+miR-29a-3p mimic group,SP+miR-29a-3p mimic+OE-NC group,SP+miR-29a-3p mimic+FSTL1-OE group(P<0.05),while miR-29a-3p level,cell survival rate,SOD and Bcl-2 levels were significantly lower(P<0.05).Compared with SP+miR-NC group,proliferation inhibition rate,apoptosis rate,TNF-α,IL-6,ROS,Bax and FSTL1 mRNA and protein expression levels were significantly lower in SP+miR-29a-3p mimic group(P<0.05),while miR-29a-3p level,cell survival rate,SOD and Bcl-2 levels were significantly higher(P<0.05).Compared with SP+miR-29a-3p mimic+OE-NC group,proliferation inhibition rate,apoptosis rate,TNF-α,IL-6,ROS,Bax,and FSTL1 mRNA and protein expression levels were significantly higher in SP+miR-29a-3p mimic+FSTL1-OE group(P<0.05),while miR-29a-3p level,cell survival rate,SOD and Bcl-2 levels were significantly lower(P<0.05).Compared with FSTL1-WT and miR-NC co-transfected cells,fluorescence enzyme activity in FSTL1-WT and miR-29a-3p mimic co-transfected group was significantly lower(P<0.05).Conclusion:miR-29a-3p may alleviate SP-induced alveolar epithelial cell injury,promote cell proliferation,inhibit cell apoptosis and inflammatory response by target down-regulating FSTL1.
王莲;杜婧慧;孙寅;蔡令辉
烟台业达医院儿科,烟台 264000||滕州市中心人民医院儿科,滕州 277599烟台业达医院儿科,烟台 264000||滕州市中心人民医院儿科,滕州 277599烟台业达医院儿科,烟台 264000||滕州市中心人民医院儿科,滕州 277599烟台业达医院儿科,烟台 264000||滕州市中心人民医院儿科,滕州 277599
医药卫生
miR-29a-3p卵泡蛋白样蛋白1肺炎链球菌肺泡上皮细胞损伤
miR-29a-3pFollistatin-like protein 1Streptococcus pneumoniaAlveolar epithelial cells injury
《中国免疫学杂志》 2026 (4)
788-793,6
山东省医药卫生科技发展计划项目(202006010355).
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