尼古丁上调Cx43促发PASMCs炎性反应致小鼠肺动脉重构的机制  

Mechanism of nicotine inducing pulmonary arterial remodeling through upregulating connexin 43 to promote inflammatory response in PASMCs in mice

作　　者：赵旭 侯晓敏[2,3] 孙琳 徐祎 董霖 江雪露 饶国娇 邹佳佳 施熠炜 秦小江[1,3,5] ZHAO Xu;HOU Xiao-min;SUN Lin;XU Yi;DONG Lin;JIANG Xue-lu;RAO Guo-jiao;ZOU Jia-jia;SHI Yi-wei;QIN Xiao-jiang(School of Public Health,Shanxi Medical University,Taiyuan 030001,China;Department of Pharmacology,Shanxi Medical University;Environmental Exposures Vascular Disease Institute,Shanxi Medical University;Academy of Medical Sciences,Shanxi Medical University;First Clinical Medical School,Shanxi Medical University)

机构地区：[1]山西医科大学公共卫生学院,太原030001 [2]山西医科大学基础医学院 [3]环境暴露血管疾病研究所,山西医科大学 [4]山西医科大学医学科学院 [5]山西医科大学第一医院

出　　处：《环境卫生学杂志》2024年第9期780-787,共8页JOURNAL OF ENVIRONMENTAL HYGIENE

基　　金：国家自然科学基金面上项目(82373622);国家自然科学基金青年基金项目(82204042);国家卫生健康委员会尘肺病重点实验室开放课题(YKFKT006、NHC202307);山西省科技创新人才团队专项项目(202304051001038);山西省留学人员科技活动择优资助项目重点项目(20220019);山西省科技合作交流专项项目(202204041101022);中央级公益性科研院所科研业务项目(2020-PT320-005)。

摘　　要：目的研究缝隙连接蛋白43(connexin43,Cx43)介导肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)炎性反应在尼古丁致小鼠肺动脉重构中的作用机制。方法5周龄雄性C57BL/6J小鼠、Tagln-Cre(+);Cx43^(flox/WT)小鼠各32只,分别随机分为对照组(灭菌注射水)、0.02、0.2和2.0 mg/(kg·d)尼古丁组,每组8只。小鼠按体重每天固定时间进行一次鼻腔滴注,持续8周。染毒结束后,通过HE染色观察各组小鼠肺动脉重构情况;采用磁性分离法培养C57BL/6J、Tagln-Cre(+);Cx43^(flox/WT)小鼠原代远端PASMCs;通过Western blot法检测小鼠PASMCs中Cx43、肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)、白细胞介素-1β(interleukin 1β,IL-1β)和白细胞介素-6(interleukin 6,IL-6)蛋白表达水平。结果与C57BL/6J小鼠对照组相比,尼古丁染毒组C57BL/6J小鼠体重总体呈现增长趋势,且随着尼古丁浓度增加,体重增长变缓。小鼠远端肺动脉出现管壁增厚、管腔狭窄等典型的动脉重构病理特征,Cx43表达上调且呈剂量依赖性关系。Tagln-Cre(+);Cx43^(flox/WT)小鼠HE染色结果显示其肺动脉管壁增厚和管腔狭窄现象有所缓解,且PASMCs中上述炎性反应因子表达减少。结论Cx43表达下调可缓解尼古丁诱导的小鼠远端PASMCs炎性反应,从而改善小鼠肺动脉重构。Objective To investigate the mechanism of the gap junction protein connexin 43(Cx43)mediating the inflammatory response of pulmonary artery smooth muscle cells(PASMCs)in nicotine-induced pulmonary artery remodeling in mice.Methods Thirty-two 5-week-old male C57BL/6J mice and Tagln-Cre(+);Cx43^(flox/WT)mice each were randomly divided into control group(sterile water for injection),0.02 mg/(kg·d)nicotine group,0.2 mg/(kg·d)nicotine,and 2.0 mg/(kg·d)nicotine group,with 8 mice in each group.The mice were treated through nasal drip according to body weight,once a day at a fixed time for continuous 8 weeks.After exposure,the mice were examined for pulmonary arterial remodeling with hematoxylin-eosin(HE)staining.The primary distal PASMCs of C57BL/6J mice and Tagln-Cre(+);Cx43^(flox/WT)mice were cultured by magnetic separation.The protein expression levels of Cx43,tumor necrosis factor-alpha,interleukin-1β,and interleukin-6 in mouse PASMCs were measured by Western blot.Results Compared with C57BL/6J mice in the control group,C57BL/6J mice in the nicotine groups showed a generally increasing trend in body weight,and the body weight growth slowed down with the increase in nicotine concentrations;the distal pulmonary arteries showed typical pathological features of arterial remodeling including wall thickening and lumen narrowing;and the expression of Cx43 was up-regulated in a dose-dependent manner.Tagln-Cre(+);HE staining results of Cx43^(flox/WT)mice showed milder wall thickening and lumen narrowing of pulmonary arteries and lower expression levels of the inflammatory factors in PASMCs.Conclusion Downregulating Cx43 can alleviate nicotine-induced inflammatory responses in distal PASMCs,thereby improving pulmonary arterial remodeling in mice.

关 键 词：尼古丁 连接蛋白43 肺动脉重构 小鼠原代肺动脉平滑肌细胞 炎性反应 

分 类 号：R12[医药卫生—环境卫生学]