摘要
目的探究无翅基因7a(Wnt7a)对牛结核分枝杆菌卡介苗(BCG)感染的肺泡上皮细胞自噬水平的调控作用。方法使用干扰Wnt7a慢病毒及BCG单独作用或共处理TC-1小鼠肺泡上皮细胞,设置小干涉RNA对照(si-NC)组、si-NC联合BCG组、Wnt7a小干涉RNA(si-Wnt7a)组和si-Wnt7a联合BCG组。Western blot法检测Wnt7a、微管相关蛋白1轻链3(LC3)、P62及自噬相关基因5(ATG5)蛋白表达水平,利用免疫荧光细胞化学染色检测LC3的分布以确定细胞自噬情况。结果与si-NC组相比,BCG感染的TC-1细胞Wnt7a、LC3和ATG5蛋白表达均升高、LC3绿色荧光斑点亮度与分布显著增加;与BCG组感染的TC-1细胞相比,si-Wnt7a联合BCG组Wnt7a、LC3、P62及ATG5蛋白表达均显著降低,LC3绿色荧光斑点亮度与分布显著降低。结论敲低Wnt7a抑制BCG诱导的肺泡上皮细胞自噬。
Objective To investigate the regulatory effect of wingless gene 7a(Wnt7a)on Bacille Calmette Guerin(BCG)induced autophagy in alveolar epithelial cells.Methods The alveolar epithelial cells of TC-1 mice were treated with interfering Wnt7a lentivirus and BCG alone or together in four groups,namely,small interfering RNA control(si-NC)group,si-NC combined with BCG group,Wnt7a small interfering RNA(si-Wnt7a)group,and si-Wnt7a combined with BCG group.The expressions of Wnt7a,microtubule associated protein 1 light chain 3(LC3),P62,and autophagy related gene 5(ATG5)were detected by Western blot analysis,and the distribution of LC3 was detected by immunofluorescence cytochemical staining.Results Compared with those in the si-NC group,Wnt7a、ATG5 and LC3 expressions increased and green fluorescent spots of LC3 significanty increased in the BCG infected TC-1 cells;the expressions of Wnt7a,LC3,P62,and ATG5 and green fluorescent spots of LC3 significantly decreased in the si-Wnt7a combined with BCG group comparing with those in the si-NC combined with BCG group.Conclusion Knocking down Wnt7a inhibits BCG induced autophagy in mouse alveolar epithelial cells.
作者
陈琪
闵思蕊
郑雪迪
聂雪伊
徐金瑞
李勇
杨易
CHEN Qi;MIN Sirui;ZHENG Xuedi;NIE Xueyi;XU Jinrui;LI Yong;YANG Yi(Key Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Westem China;School of Life Sciences,Ningxia University,Yinchuan 750021,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2023年第2期97-102,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(31960712,31960700)。
作者简介
同为第一作者:陈琪(1995-),女,甘肃平凉人,硕士研究生,Tel:18195125716,E-mail:3192216154@qq.com;同为第一作者:闵思蕊;通讯作者:杨易,E-mail:yangyi@nxu.edu.cn。
