LncRNA GAS5靶向调控miR-29a-3p对肺炎链球菌诱导的人肺泡上皮细胞增殖、凋亡的影响OA
Effects of LncRNA GAS5 on proliferation and apoptosis of human alveolar epithelial cells induced by Streptococcus pneumoniae through targeted regulation of miR-29a-3p
目的:探讨LncRNA GAS5靶向调控miR-29a-3p对肺炎链球菌诱导的人肺泡上皮细胞增殖、凋亡的影响.方法:当A549细胞达到80%汇合时,进行实验分组:对照组(正常培养)、感染(Infection,1×108 CFU/mL肺炎链球菌孵育)组、Infec-tion+阴性对照(pcDNA)组、Infection+GAS5过表达(pcDNA GAS5)组、Infection+miR-29a-3p抑制剂(inhibitor)组、Infection+阴性对照(inhibitor NC)组、Infection+pcDNA GAS5+阴性对照(miR NC)组、Infection+pcDNA GAS5+miR-29a-3p组,利用脂体转染法将上述质粒或寡核苷酸转染至A549细胞,随后均以1×108 CFU/mL肺炎链球菌孵育48 h,随后检测GAS5、miR-29a-3p表达、细胞增殖及相关蛋白(Ki67、PCNA)表达、凋亡及相关蛋白(Bax、Bcl-2)表达、上清液中炎症因子IL-6、IL-1β、IL-10水平以及GAS5、miR-29a-3p的靶向关系.结果:与对照组相比,感染组GAS5表达、增殖率、IL-10水平、Ki67、PCNA、Bcl-2表达显著降低,但miR-29a-3p、凋亡率、IL-6、IL-1β水平、Bax表达显著升高(P<0.05);与Infection+pcDNA组 相 比,Infection+pcDNA GAS5组GAS5表达、增殖率、IL-10水平、Ki67、PCNA、Bcl-2表达显著升高,但miR-29a-3p、凋亡率、IL-6、IL-1β、水平、Bax表达显著降低(P<0.05);与Infection+inhibitor NC组相比,Infection+miR-29a-3p inhibitor组增殖率、IL-10水平、Ki67、PCNA、Bcl-2表达显著增加,但miR-29a-3p、凋亡率、IL-6、IL-1β水平、Bax表达显著降低(P<0.05),GAS5表达差异无统计学意义(P>0.05);与Infection+pcDNA GAS5+miR NC组相比,Infection+pcDNA GAS5+miR-29a-3p组增殖率、IL-10水平、Ki67、PCNA、Bcl-2表达显著降低,但miR-29a-3p、凋亡率、IL-6、IL-1β水平、Bax表达显著升高(P<0.05),GAS5表达差异无统计学意义(P>0.05).GAS5、miR-29a-3p为负靶向调节关系.结论:上调LncRNA GAS5靶向调控miR-29a-3p减少肺炎链球菌诱导的人肺泡上皮细胞增殖,促进其凋亡.
Objective:To investigate the effects of LncRNA GAS5 on the proliferation and apoptosis of human alveolar epithe-lial cells induced by Streptococcus pneumoniae through targeted regulation of miR-29a-3p.Methods:When A549 cells reached 80%confluence,experimental groups were performed:Control group(normal culture),infection(1×108 CFU/mL Streptococcus pneumoniae incubation)group,infection+negative control(pcDNA)group,infection+GAS5 overexpression(pcDNA GAS5)group,infection+miR-29a-3p inhibitor(inhibitor)group,infection+negative control(inhibitor NC)group,infection+pcDNA GAS5+negative control(miR NC)group,and infection+pcDNA GAS5+miR-29a-3p group.The above plasmids or oligonucleotides were transfected into A549 cells using liposome transfection method,and then incubated with 1×108 CFU/mL Streptococcus pneumoniae for 48 hours.Subsequently,GAS5,miR-29a-3p expression,cell proliferation and related protein(Ki-67,PCNA)expression,apoptosis and related protein(Bax,Bcl-2)expression,inflammatory cytokines IL-6,IL-1β,IL-10 levels in the supernatant,and the targeting relationship between GAS5 and miR-29a-3p were detected.Results:Compared with the control group,the expression of GAS5,proliferation rate,IL-10 level,Ki67,PCNA and Bcl-2 in the infection group were obviously lower,while miR-29a-3p,apoptosis rate,IL-6,IL-1β level,and Bax expression were obviously higher(P<0.05).Compared with the infection+pcDNA group,the GAS5 expression,proliferation rate,IL-10 level,Ki67,PCNA and Bcl-2 expressions were obviously higher in the infection+pcDNA GAS5 group,but miR-29a-3p,apoptosis rate,IL-6,IL-1β level,and Bax expression were obviously lower(P<0.05).Compared with the infection+inhibitor NC group,the pro-liferation rate,IL-10 level,Ki67,PCNA and Bcl-2 expressions were obviously higher in the infection+miR-29a-3p inhibitor group.However,miR-29a-3p,apoptosis rate,IL-6,IL-1β levels and Bax expression were obviously lower(P<0.05),while GAS5 expres-sion showed no obvious difference(P>0.05).Compared with the infection+pcDNA GAS5+miR NC group,the proliferation rate,IL-10 level,Ki67,PCNA and Bcl-2 expressions in the infection+pcDNA GAS5+miR-29a-3p group were obviously lower,but miR-29a-3p,apoptosis rate,IL-6,IL-1β levels and Bax expression were obviously higher(P<0.05),while there was no obvious difference in GAS5 expression(P>0.05).GAS5 and miR-29a-3p had a negative targeted regulatory relationship.Conclusion:Up-regulation of Ln-cRNA GAS5 reduces the proliferation and promotes apoptosis of human alveolar epithelial cells induced by Streptococcus pneumoniae by targeting miR-29a-3p.
李群;蔡明丹;张萍萍;黄志心;陈海燕;谢丛华
武汉市第三医院儿科,武汉 430060武汉市第三医院儿科,武汉 430060武汉市第三医院儿科,武汉 430060武汉市第三医院儿科,武汉 430060武汉市第三医院儿科,武汉 430060武汉大学中南医院呼吸内科,武汉 430000
医药卫生
LncRNA GAS5miR-29a-3p肺炎链球菌人肺泡上皮细胞增殖凋亡
LncRNA GAS5miR-29a-3pStreptococcus pneumoniaeHuman alveolar epithelial cellsProliferationApoptosis
《中国免疫学杂志》 2026 (8)
1803-1808,6
湖北省卫生健康委员会联合基金项目(WJ2019H002).
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