宣肺平喘胶囊基于p38磷酸化途径对COPD自噬影响研究  

Xuanfei Pingchuan capsules ameliorate autophagy in chronic obstructive pulmonary disease by inhibiting p38 phosphorylation

作　　者：张晔 薛晓明[2] 孟丽红 李豪 李典 乔文晓 ZHANG Ye;XUE Xiaoming;MENG Lihong;LI Hao;LI Dian;QIAO Wenxiao(Shanxi University of Chinese Medicine,Taiyuan 030002,Shanxi,China;Shanxi Traditional Chinese Medicine Hospital,Taiyuan 030012,Shanxi,China)

机构地区：[1]山西中医药大学,山西太原030002 [2]山西省中医院,山西太原030012

出　　处：《现代中西医结合杂志》2022年第24期3366-3371,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：山西省科技厅优秀人才科技创新项目(201806D211003)。

摘　　要：目的 探讨宣肺平喘胶囊是否通过抑制p38磷酸化而影响香烟烟雾提取物诱导的支气管上皮细胞自噬。方法 将支气管上皮细胞分为空白组、香烟烟雾提取物组、宣肺平喘低剂量组、宣肺平喘中剂量组、宣肺平喘高剂量组。以香烟烟雾提取物诱导支气管上皮细胞建立慢性阻塞性肺疾病(COPD)细胞模型,空白组以空白血清培养,宣肺平喘各组加入宣肺平喘含药血清培养。CCK-8法检测细胞活力,流式细胞仪检测细胞凋亡率,免疫荧光显微镜观察自噬情况,Western blot法检测细胞中p38、p-p38、LC3-Ⅰ、LC3-Ⅱ、Beclin 1蛋白表达情况,RT-PCR法检测细胞中LC3-Ⅰ、LC3-Ⅱ、Beclin 1 mRNA表达情况。结果 与空白组比较,香烟烟雾提取物组细胞活力均显著降低(P<0.05),细胞凋亡率、自噬水平和p-p38、LC3-Ⅰ、LC3-Ⅱ、Beclin 1蛋白表达量及LC3-Ⅰ、LC3-Ⅱ、Beclin 1 mRNA表达量均显著增高(P均<0.05);与香烟烟雾提取物组比较,宣肺平喘各组细胞活力均显著增加(P均<0.05),细胞凋亡率、自噬水平和p-p38、LC3-Ⅰ、LC3-Ⅱ、Beclin 1蛋白表达量及LC3-Ⅰ、LC3-Ⅱ、Beclin 1 mRNA表达量均显著降低(P均<0.05),宣肺平喘高剂量组各指标改变更为显著。结论 宣肺平喘胶囊能够显著抑制香烟烟雾提取物诱导的支气管上皮细胞自噬,从而减少细胞凋亡,增强细胞活力,p38磷酸化在这一过程中发挥了关键作用。Objective It is to investigate whether Xuanfei Pingchuan capsules affect cigarette smoke extract-induced autophagy in bronchial epithelial cells by inhibiting p38 phosphorylation. Methods The bronchial epithelial cells were divided into blank group, cigarette smoke extract group, Xuanfei Pingchuan low dose group, Xuanfei Pingchuan medium dose group and Xuanfei Pingchuan high dose group. The bronchial epithelial cells were induced by cigarette smoke extract to establish chronic obstructive pulmonary disease(COPD) cell models, and the blank group was cultured with blank serum, and each Xuanfei Pingchuan group was cultured with corresponding Xuanfei Pingchuan drug-containing serum. The cell viability was detected by CCK-8 method, apoptosis rate was detected by flow cytometry, autophagy was observed by immunofluorescence microscopy, the expression of p38, p-p38, LC3-Ⅰ, LC3-Ⅱ, Beclin 1 protein in cells was detected by Western blot, and the expression of LC3-Ⅰ, LC3-Ⅱ, Beclin 1 mRNA in cells was detected by RT-PCR. Results Compared with the blank group, the cell viability of the cigarette smoke extract group was significantly decreased(P<0.05), and the apoptosis rate and the level of autophagy in cells, the expression of p-p38, LC3-Ⅰ, LC3-Ⅱ, Beclin 1 protein and the expression of LC3-Ⅰ, LC3-Ⅱ, Beclin 1 mRNA were significantly increased(all P<0.05). Compared with the cigarette smoke extract group, the cell viability was significantly increased, and the apoptosis rate, autophagy level and p-p38, LC3-Ⅰ, LC3-Ⅱ, Beclin 1 protein expression and LC3-Ⅰ, LC3-Ⅱ, Beclin 1 mRNA expression were significantly decreased in every Xuanfei Pingchuan group(all P<0.05), and the changes of each index were more significant in the Xuanfei Pingchuan high dose group. Conclusion Xuanfei Pingchuan capsule could significantly inhibit cigarette smoke extract-induced autophagy in bronchial epithelial cells, thereby reducing apoptosis and enhancing cell viability, with p38 phosphorylation playing a key role in this process.

关 键 词：慢性阻塞性肺疾病 宣肺平喘胶囊 香烟烟雾提取物 自噬 p38磷酸化 

分 类 号：R563[医药卫生—呼吸系统] R329.28[医药卫生—内科学]