清金化痰汤通过Bax/Bcl-2信号通路抑制气道上皮细胞自噬  

Research on Qingjin Huatan Decoction(清金化痰汤)Inhibiting Autophagy of Airway Epithelial Cells through Bax/Bcl-2 Signal Pathway

作　　者：裴凯 洪妹 彭思敏 王林 张艳艳 许光兰[2] PEI Kai;HONG Mei;PENG Simin;WANG Lin;ZHANG Yanyan;XU Guanglan(Guangxi University of Traditional Chinese Medicine,Nanning 530001,Guangxi,China;The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《辽宁中医药大学学报》2023年第11期41-45,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81760848,82160889);广西高校中青年教师科研基础能力提升项目(2021KY310);广西医疗卫生重点培育学科建设项目。

摘　　要：目的探讨清金化痰汤通过Bax、Bcl-2通路对脂多糖(LPS)及香烟烟雾(CSE)干预的大鼠原代气道上皮细胞自噬的影响。方法将大鼠按照随机抽签法分为正常组、模型组、阳性组、中药组,每组10只,采用气管滴注LPS+烟熏制备AECOPD大鼠模型,并且对气道上皮细胞(正常和痰热郁肺型AECOPD大鼠)进行原代提取。制备清金化痰汤以及罗红霉素含药血清作用于上皮细胞。MTT检测清金化痰汤含药血清干预后细胞增殖率;流式细胞术检测清金化痰汤含药血清干预后细胞凋亡率;AD-mcherry-EGFP-lc3腺病毒转染检测气道上皮细胞内自噬体含量;Western Blot检测气道上皮细胞内凋亡蛋白Bax、Bcl-2表达。结果MTT结果显示:正常组与给药组跟模型组比较差异均有统计学意义(P<0.05),流式结果显示:与正常组相比,模型组细胞活力明显降低(P<0.05);与模型组比较,在使用含药血清后,细胞活力具有明显提升(P<0.05);腺病毒转染显示:模型组与正常组相比大鼠气道上皮细胞自噬体数量增多(P<0.05);与模型组比较,经过含药血清处理后,AECOPD大鼠的气道上皮细胞内的自噬体明显降低(P<0.05);Western Blot结果显示:与正常组比较,模型组凋亡蛋白表达明显升高(P<0.05);与模型组比较,清金化痰汤药物干预组凋亡蛋白Bax表达下降(P<0.05);清金化痰汤高剂量组与罗红霉素组比较无统计学意义。结论清金化痰汤能抑制气道上皮细胞中炎症反应和凋亡的机制可能与调节自噬体有关。Objective To investigate the effect of Qingjin Huatan Decoction(清金化痰汤)on autophagy of primary airway epithelial cells in rats interfered by lipopolysaccharide(LPS)through Bax and Bcl-2 pathways.Methods The rats were randomly divided into normal group,model group,positive group and traditional Chinese medicine group,with 10 rats in each group.The AECOPD rat model was prepared by intratracheal instillation of LPS+smoking,and the airway epithelial cells(normal and phlegm heat stagnation lung type AECOPD rats)were extracted in primary culture.Qingjin Huatan Decoction and roxithromycin containing serum were prepared to act on epithelial cells.MTT was used to detect the cell proliferation rate after the intervention of serum containing Qingjin Huatan Decoction;The apoptosis rate was detected by flow cytometry after the intervention of serum containing Qingjin Huatan Decoction;AD-mcherry-EGFP-lc3 adenovirus transfection was used to detect the content of autophage in airway epithelial cells;Western blot was used to detect the expression of Bax and Bcl-2 in airway epithelial cells.Results MTT results showed that there were significant differences between the normal group and the drug administration group and the model group(P<0.05).Flow cytometry results showed that the cell viability of the model group was significantly lower than that of the normal group(P<0.05);Compared with the model group,the cell viability was significantly improved after the use of drug containing serum(P<0.05);Adenovirus transfection showed that the number of autophages in airway epithelial cells of rats in the model group increased compared with that in the normal group(P<0.05);Compared with the model group,after treatment with drug containing serum,the autophage in airway epithelial cells of AECOPD rats decreased significantly(P<0.05);Western blot showed that the expression of apoptosis protein in model group was significantly higher than that in normal group(P<0.05);Compared with the model group,the expression of apoptosis protein Bax

关 键 词：清金化痰汤 慢性阻塞性肺疾病 自噬体 凋亡 炎症 

分 类 号：R285.5[医药卫生—中药学]