[1]魏士刚,马雪梅,宿海峰.长链非编码RNA SIL对肺炎链球菌感染的肺上皮细胞增殖、凋亡及转化生长因子β1/Smads通路的影响[J].陕西医学杂志,2023,52(6):645-650.[doi:DOI:10.3969/j.issn.1000-7377.2023.06.003]
　WEI Shigang,MA Xuemei,SU Haifeng.Effects of lncRNA SIL on proliferation,apoptosis and TGF-β1/Smads pathway of lung epithelial cells infected with Streptococcus pneumoniae[J].,2023,52(6):645-650.[doi:DOI:10.3969/j.issn.1000-7377.2023.06.003]
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长链非编码RNA SIL对肺炎链球菌感染的肺上皮细胞增殖、凋亡及转化生长因子β1/Smads通路的影响
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年6期
页码:
645-650
栏目:
基础研究
出版日期:
2023-06-05

文章信息/Info

Title:
Effects of lncRNA SIL on proliferation,apoptosis and TGF-β1/Smads pathway of lung epithelial cells infected with Streptococcus pneumoniae
作者:
魏士刚1; 马雪梅2; 宿海峰1
(1.北京航天总医院,北京 100076; 2.佳木斯大学附属第一医院,黑龙江 佳木斯 154002)
Author(s):
WEI Shigang; MA Xuemei; SU Haifeng
(Beijing Aerospace General Hospital,Beijing 100076,China)
关键词:
肺炎链球菌感染;  长链非编码RNA SIL;  肺上皮细胞;  细胞增殖;  细胞凋亡;  转化生长因子β1/Smads通路
Keywords:
Streptococcus pneumoniae infection;  Long non-coding RNA SIL;  Lung epithelial cells;  Cell proliferation;  Apoptosis;  TGF-β1/Smads pathway
分类号:
R 563.1
DOI:
DOI:10.3969/j.issn.1000-7377.2023.06.003
文献标志码:
A
摘要:
目的:探讨长链非编码RNA(lncRNA)SIL对肺炎链球菌感染的肺上皮细胞细胞增殖、凋亡及转化生长因子β1(TGF-β1)/Smads通路的影响。方法:运用反转录PCR(RT-PCR)检测lncRNA SIL在人肺泡上皮细胞株A549、HEPApiC、肺泡Ⅱ型细胞(AECⅡ)以及被肺炎链球菌R6感染的A549、HEPApiC、AECⅡ细胞的表达,显示其表达在R6感染的A549细胞中最高,故应用A549细胞进行后续实验。将A549细胞随机分为A549组(常规培养)、A549+R6组(用R6感染A549细胞)、lncRNA SIL mimic组(在A549细胞中转染lncRNA SIL mimic,并用R6感染细胞)和lncRNA SIL siRNA组(在A549细胞中转染lncRNA SIL siRNA,并用R6感染细胞)。采用CCK-8法检测各组细胞增殖情况。采用流式细胞仪检测各组细胞凋亡情况。采用免疫印迹法检测TGF-β1/Smads通路相关蛋白表达,并设立TGF-β1/Smads通路抑制剂组进行对照。采用酶联免疫吸附试验(ELISA)检测各组炎症因子表达。结果:与A549组比较,A549+R6组lncRNA SIL表达升高(P<0.05)。与A549+R6组比较,lncRNA SIL mimic组lncRNA SIL表达升高,lncRNA SIL siRNA组表达降低(均P<0.05)。与lncRNA SIL mimic组比较,lncRNA SIL siRNA组lncRNA SIL表达降低(P<0.05)。与A549组比较,A549+R6组不同时间细胞增殖率降低(均P<0.05)。与A549+R6组比较,lncRNA SIL mimic组不同时间细胞增殖率降低,lncRNA SIL siRNA组不同时间细胞增殖率升高(均P<0.05)。与A549组比较,A549+R6组细胞凋亡率增加(P<0.05)。与A549+R6组比较,lncRNA SIL mimic组细胞凋亡率增加,lncRNA SIL siRNA组细胞凋亡率降低(均P<0.05)。与A549组比较,A549+R6组TGF-β1、Smad 2、Smad 3蛋白表达升高,Smad 6、Smad 7蛋白表达降低(均P<0.05)。与A549+R6组比较,lncRNA SIL mimic组TGF-β1、Smad 2、Smad 3蛋白表达升高,Smad 6、Smad 7蛋白表达降低(均P<0.05)。与A549+R6组比较,lncRNA SIL siRNA组TGF-β1、Smad 2、Smad 3蛋白表达降低,Smad 6、Smad 7蛋白表达升高(均P<0.05)。lncRNA SIL siRNA组与抑制剂组各指标比较差异无统计学意义(均P>0.05)。与A549组比较,A549+R6组肿瘤坏死因子-α(TNF-α)、白介素细胞-1β(IL-1β)、IL-6水平升高(均P<0.05)。与A549+R6组比较,lncRNA SIL mimic组TNF-α、IL-1β、IL-6水平升高,lncRNA SIL siRNA组TNF-α、IL-1β、IL-6水平降低(均P<0.05)。结论:肺炎链球菌感染后,沉默lncRNA SIL可逆转肺上皮细胞凋亡增加及增殖减少的情况,并抑制TGF-β1/Smads通路激活,减少炎症因子释放,从而抑制炎性反应。
Abstract:
Objective:To investigate the effects of long non-coding RNA(lncRNA)SIL on the proliferation,apoptosis and transforming growth factor-β1(TGF-β1)/Smads signaling pathway of lung epithelial cells infected with Streptococcus pneumoniae.Methods:RT-PCR was used to detect the expression of lncRNA SIL in human alveolar epithelial cell lines A549,HEPApiC and AECⅡ,and A549,HEPApiC,and AECⅡ cells infected with Streptococcus pneumoniae R6.The expression of lncRNA SIL was the highest in A549 cells infected with R6.Therefore,A549 cells were used for subsequent experiments.A549 cells were randomly divided into A549 group(conventional culture),A549+R6 group(cells were infected with R6),lncRNA SIL mimic group(A549 cells were transfected with lncRNA SIL mimic and infected with R6)and lncRNA SIL siRNA group(A549 cells were transfected with lncRNA SIL siRNA and infected with R6).Cell proliferation was detected by CCK-8 assay.Flow cytometry was used to detect cell apoptosis in each group.Western blot was used to detect the expression of TGF-β1/Smads pathway-related proteins,and the TGF-β1/Smads pathway inhibitor group was set up as a control.ELISA was used to detect the expression of inflammatory factors in each group.Results:Compared with A549 group,lncRNA SIL expression was significantly increased in A549+R6 group(P<0.05).Compared with the A549+R6 group,the expression of lncRNA SIL was increased in the lncRNA SIL mimic group and decreased in the lncRNA SIL siRNA group(both P<0.05).Compared with the lncRNA SIL mimic group,the expression of lncRNA SIL in the lncRNA SIL siRNA group was decreased(P<0.05).Compared with A549 group,the cell proliferation rate of A549+R6 group was decreased at different time points(all P<0.05).Compared with the A549+R6 group,the cell proliferation rate in the lncRNA SIL mimic group was decreased at different time points,and the cell proliferation rate in the lncRNA SIL siRNA group was increased at different time points(all P<0.05).Compared with A549 group,the apoptosis rate of A549+R6 group was increased(P<0.05).Compared with the A549+R6 group,the apoptosis rate of lncRNA SIL mimic group was increased,and the apoptosis rate of lncRNA SIL siRNA group was decreased(both P<0.05).Compared with the A549 group,the protein expressions of TGF-β1,Smad 2 and Smad 3 were increased,and the protein expressions of Smad 6 and Smad 7 were decreased in the A549+R6 group(all P<0.05).Compared with the A549+R6 group,the expressions of TGF-β1,Smad 2 and Smad 3 proteins in the lncRNA SIL mimic group were increased,while the expressions of Smad 6 and Smad 7 proteins were decreased(all P<0.05).Compared with the A549+R6 group,the protein expressions of TGF-β1,Smad 2 and Smad 3 were decreased,and the protein expressions of Smad 6 and Smad 7 were increased in the lncRNA SIL siRNA group(all P<0.05).There were no significant differences in all indicators between lncRNA SIL siRNA group and inhibitor group(all P>0.05).Compared with A549 group,the levels of TNF-α,IL-1β and IL-6 in A549+R6 group were increased(all P<0.05).Compared with the A549+R6 group,the levels of TNF-α,IL-1β and IL-6 were increased in the lncRNA SIL mimic group,and decreased in the lncRNA SIL siRNA group(all P<0.05).Conclusion:Silencing lncRNA SIL can reverse the increase of apoptosis and decrease of proliferation of lung epithelial cells after Streptococcus pneumoniae infection,inhibit the activation of TGF-β1/Smads pathway,reduce the release of inflammatory factors,thereby inhibiting the inflammatory response.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81370287); 黑龙江省卫生和计划生育委员会科研课题(2017-399)
更新日期/Last Update: 2023-06-05