Zhang Ye, Xue Xiaoming, Meng Lihong, Li Dian, Qiao Wenxiao, Wang Jinyun, Xie Di
Graduate School, Shanxi University of Chinese Medicine, Taiyuan, China.
Front Pharmacol. 2023 May 18;14:1123882. doi: 10.3389/fphar.2023.1123882. eCollection 2023.
Autophagy plays an important role in the occurrence and development of chronic obstructive pulmonary disease (COPD). We evaluated the effect of Xuanfei Pingchuan capsule (XFPC) on autophagy-related genes of COPD by a bioinformatics analysis and experimental verification. The best treatment duration was screened by CCK8 assays. HBE cells were divided into three groups: blank, CSE and XFPC. After intervened by XFPC, HBE cells were collected and sent to Shenzhen Huada Gene Company for transcriptome sequencing. Subsequently, differential expression analyses, target gene prediction, and function enrichment analyses were carried out. Expression changes were verified in HBE cells by real-time Quantitative PCR (RT-qPCR) and western blotting (WB). The result of differential expression analysis displayed that 125 target genes of HBE cells were mainly related to mitogen-activated protein kinase (MKK) binding, interleukin 33 binding, 1-Pyrroline-5-carboxylate dehydrogenase activity, and the mitogen-activated protein kinase (MAPK) signal pathway. Among the target genes, the core genes related to autophagy obtained by maximum neighborhood component algorithm were , , , , and . RT-qPCR and WB methods were used to verify the result, it showed similar expression changes in , , , and in bronchial epithelial cells to those in the bioinformatics analysis. Through transcriptome sequencing and validation analysis, we predicted that CSF1, MAPK9, MAP3K7, and AKT3 may be the potential autophagy-related genes that play an important role in the pathogenesis of COPD. XFPC may regulate autophagy by down-regulating the expression of CSF1, MAPK9, MAP3K7, and AKT3, thus achieving the purpose of treating chronic obstructive pulmonary disease.
自噬在慢性阻塞性肺疾病（COPD）的发生发展中起重要作用。我们通过生物信息学分析和实验验证评估了宣肺平喘胶囊（XFPC）对COPD自噬相关基因的影响。通过CCK8试验筛选出最佳治疗时长。将人支气管上皮（HBE）细胞分为三组：空白组、香烟烟雾提取物（CSE）组和XFPC组。XFPC干预后，收集HBE细胞并送至深圳华大基因公司进行转录组测序。随后，进行差异表达分析、靶基因预测和功能富集分析。通过实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法（WB）在HBE细胞中验证表达变化。差异表达分析结果显示，HBE细胞的125个靶基因主要与丝裂原活化蛋白激酶（MKK）结合、白细胞介素33结合、1-吡咯啉-5-羧酸脱氢酶活性以及丝裂原活化蛋白激酶（MAPK）信号通路有关。在这些靶基因中，通过最大邻域成分算法获得的与自噬相关的核心基因是 、 、 、 和 。采用RT-qPCR和WB方法验证结果，结果显示支气管上皮细胞中 、 、 和 的表达变化与生物信息学分析中的相似。通过转录组测序和验证分析，我们预测集落刺激因子1（CSF1）、丝裂原活化蛋白激酶9（MAPK9）、丝裂原活化蛋白激酶激酶激酶7（MAP3K7）和蛋白激酶B（AKT3）可能是在COPD发病机制中起重要作用的潜在自噬相关基因。XFPC可能通过下调CSF1、MAPK9、MAP3K7和AKT3的表达来调节自噬，从而达到治疗慢性阻塞性肺疾病的目的。
