摘要
目的:探讨苯并(a)芘(BaP)促凋亡诱导哮喘气道上皮细胞损伤,及芪仙汤的保护机制。方法:以16HBE细胞株为体外研究对象,体内研究采用OVA诱导哮喘小鼠模型,BaP滴鼻诱导哮喘气道损伤。MTT检测16HBE细胞存活率;Annexin V-FITC双染检测16HBE细胞凋亡;Western Blot检测16HBE细胞及肺组织Bcl-2、Bax蛋白及16HBE细胞HO-1蛋白表达水平;qPCR检测16HBE细胞Bcl-2、Bax mRNA表达水平;DCFH-DA探针检测16HBE细胞活性氧(ROS)水平。结果:与对照组比较,BaP干预后可增加16HBE细胞抑制率,凋亡水平,ROS水平及HO-1蛋白表达水平,升高16HBE细胞Bax蛋白及mRNA水平,降低Bcl-2蛋白及mRNA水平(P<0.05);与哮喘组比较,BaP干预后可进一步加重哮喘小鼠肺组织气道炎症与气道壁损伤,进一步上调哮喘小鼠肺组织Bax蛋白表达,下调Bcl-2蛋白表达(P<0.05);与BaP组比较,芪仙汤提取物可降低16HBE细胞抑制率,降低16HBE细胞凋亡及ROS水平,上调HO-1蛋白表达,下调小鼠肺组织及16HBE细胞Bax蛋白表达水平及增加Bcl-2蛋白表达水平(P<0.05)。结论:芪仙汤增加HO-1表达提高细胞抗氧化水平,逆转BaP诱导的哮喘气道上皮细胞凋亡。
Objective: To investigate the mechanism of benzo(a)pyrene promoting apoptosis-induced airway epithelial cell damage in asthma and the protective mechanism of Qixian Decoction. Methods: 16HBE cell line was used as the object of in vitro study. The OVA-induced asthma mouse model was used as the object of in vivo study, and benzo(a)pyrene nasal drip induced asthma airway injury. 16HBE cells viability were detected by MTT. 16HBE cells apoptosis were detected by Annexin V-FITC double staining. Western Blot was used to detected the protein expression levels of Bcl-2 and Bax in 16HBE cells and lung tissue,and HO-1 protein expression in 16HBE cells. qPCR was used to detected the expression levels of Bcl-2 and Bax mRNA in 16HBE cells. Cell ROS levels were detected by DCFH-DA probe. Results: Compared with the control group, BaP intervention increased the inhibition rate, apoptosis level, ROS level and HO-1 protein expression level of 16HBE cells, increased Bax protein and mRNA levels, and decreased Bcl-2 protein and mRNA levels in 16HBE cells(P<0.05). Compared with the asthma group, BaP intervention further aggravated the airway inflammation and damaged airway wall in the lung tissue of asthmatic mice, further up-regulated the expression level of Bax protein level in the lung tissue of asthmatic mice, and reduced the expression level of Bcl-2 protein(P<0.05). Compared with the BaP group, Qixian Decoction extract increased the inhibition rate of 16HBE cells,reduced the apoptosis and ROS levels of 16HBE cells, increased the expression level of HO-1 protein, and reduced the expression level of Bax protein in lung tissue in mice and 16HBE cells, and increased the expression level of Bcl-2 protein(P<0.05).Conclusion: Qixian Decoction increases the expression of HO-1 and increases the antioxidant level of cells, and reverses the apoptosis of asthmatic airway epithelial cells induced by benzo(a)pyrene.
作者
汤玲玲
王雄彪
孙宪泓
刘丽
史锁芳
TANG Ling-ling;WANG Xiong-biao;SUN Xian-hong;LIU Li;SHI Suo-fang(Nanjing University of Chinese Medicine,Nanjing 210029,China;Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2022年第9期5356-5360,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金青年科学基金项目(No.82004265),国家自然科学基金面上项目(No.81774267)。
关键词
哮喘
凋亡
氧化应激
苯并(A)芘
芪仙汤
Asthma
Apoptosis
Oxidant stress
Benzo(a)pyrene
Qixian Decoction
