白细胞介素1β基因沉默对烟草烟雾提取物诱发血管平滑肌细胞凋亡和炎性反应的影响  

Effects of IL-1βgene silencing on apoptosis and inflammatory response of VSMC induced by cigarette smoke extract

作　　者：姜红峰[1] 郭张强 吴娟 曾昆[1] 汤海艳 谭含璇 闵瑞 黄蔡华[1] 张洁 彭绍蓉 Jiang Hongfeng;Guo Zhangqiang;Wu Juan;Zeng Kun;Tang Haiyan;Tan Hanxuan;Min Rui;Huang Caihua;Zhang Jie;Peng Shaorong(Department of Geriatrics,Wuhan Fourth Hospital·Pu'ai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430033,Hubei Province,China)

机构地区：[1]武汉市第四医院华中科技大学同济医学院附属普爱医院老年病科,430033 [2]武汉市第四医院急诊内科,430033

出　　处：《中华老年心脑血管病杂志》2022年第8期866-870,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：武汉市医学科研项目(WX19D63)。

摘　　要：目的探讨白细胞介素(IL)-1β基因沉默和过表达对烟草烟雾提取物(CSE)诱导血管平滑肌细胞凋亡和炎性反应的影响。方法根据IL-1βmRNA编码序列设计并合成小干扰RNA(siRNA)和过表达质粒,分别转染大鼠胸主动脉平滑肌细胞A7r5,实验分为对照组、模型组(CSE干预)、沉默组(siRNA-IL-1β)、沉默空载组(siRNA-EV)、过表达组(OvExp-IL-1β)、过表达空载组(OvExp-EV)。采用CSE干预除对照组的其他5组细胞。qRT-PCR检测细胞中IL-1βmRNA表达。流式细胞术检测细胞凋亡率。Western blot检测细胞中B淋巴细胞瘤2基因(Bcl-2)、Bax、TNF-α、IL-6和IL-8表达。结果与对照组比较,模型组细胞凋亡率、Bax、IL-1βmRNA、TNF-α、IL-6和IL-8表达明显升高,Bcl-2表达明显降低(P<0.01)。与模型组比较,沉默组细胞凋亡率、Bax、IL-1βmRNA、TNF-α、IL-6和IL-8表达明显降低,Bcl-2表达明显升高(P<0.01);过表达组细胞凋亡率、Bax、IL-1βmRNA、TNF-α、IL-6和IL-8表达明显升高,Bcl-2表达明显降低,差异有统计学意义(P<0.01)。与沉默空载组比较,沉默组细胞凋亡率、IL-1βmRNA、TNF-α、IL-6和IL-8明显降低(P<0.05,P<0.01)。与沉默空载组比较,沉默组细胞中Bax表达明显降低[(0.35±0.02)μg/ml vs(0.42±0.02)μg/ml,P<0.01],Bcl-2表达明显升高[(0.45±0.02)μg/ml vs(0.37±0.01)μg/ml,P<0.01]。结论CSE可诱导大鼠胸主动脉平滑肌细胞凋亡和炎性反应,IL-1β基因沉默可抑制CSE诱导的细胞凋亡和炎性反应;反之,IL-1β基因过表达可进一步促进细胞凋亡和炎症水平。Objective To investigate the effects of silencing and overexpression of IL-1βgene on the apoptosis and inflammatory response of rat vascular smooth muscle cells induced by CSE.Methods Based on the coding sequence of IL-1βmRNA,its interference siRNA was designed and synthesized and overexpression plasmid was constructed,and then were transfected into rat thoracic aortic smooth muscle A7r5 cells respectively.The cells were divided into:control group,model group,siRNA-IL-1βgroup,siRNA-EV group,OvExp-IL-1βgroup and OvExp-EV group.CSE was used to intervene the other 5 groups of cells except the control group.The mRNA expression of IL-1βwas detected by qRT-PCR.Cell apoptotic rate was measured with flow cytometry.The protein levels of Bcl-2,Bax,TNF-α,IL-6 and IL-8 were detected by Western blotting.Results Compared with the control group,the apoptotic rate,IL-1βmRNA level and protein levels of Bax,TNF-α,IL-6 and IL-8 were significantly increased,while the protein level of Bcl-2 was obviously decreased in the model group.Sliencing of IL-1βresulted in notably decreased apoptotic rate,IL-1βmRNA level and protein levels of Bax,TNF-α,IL-6 and IL-8,but increased Bcl-2 pro-tein level when compared with the conditions in the model group.However,overexpression of IL-1βreversed the above results(P<0.01).Compared with the siRNA-EV group,the apoptotic rate,IL-1βmRNA level,and TNF-α,IL-6 and IL-8 levels were significantly decreased in the siRNA-IL-1βgroup(P<0.05,P<0.01).The expression of Bax was notably decreased(0.35±0.02μg/ml vs 0.42±0.02μg/ml,P<0.01)and the expression of Bcl-2(0.45±0.02μg/ml vs 0.37±0.01μg/ml,P<0.01)was remarkably elevated in the siRNA-IL-1βgroup than the siRNA-EV group.Conclusion CSE can induce apoptosis and inflammatory response in rat thoracic aortic smooth muscle cells,and IL-1βgene silencing can inhibit CSE-induced apoptosis and inflammatory response.On the contrary,its overexpression can further promote cell apoptosis and inflammation.

关 键 词：白细胞介素1Β 基因沉默 烟草 烟雾 肌 平滑 血管 细胞凋亡 

分 类 号：R163[医药卫生—公共卫生与预防医学]