目的 探讨汉黄芩素通过调控肺上皮间质形成因子的表达，治疗肺炎支原体肺炎的作用机制。方法 运用超高效液相色谱-质谱联用(UPLC-Q-TOF-MS)技术检测芩百清肺浓缩丸中的活性成分汉黄芩素，通过Biacore垂钓技术将其与转化生长因子-β1(transforming growth factor-β1，TGF-β1)进行结合，动力学分析验证其结合的特异性。采用Tripos Sybyl-X2.1.1以及Discovery Studio2016软件对汉黄芩素和TGF-β1进行分子对接，对药物分子与受体蛋白的结合模式进行可视化分析。通过动物实验制作小鼠肺部病理组织切片，进行苏木精-伊红(hematoxylin-eosin，HE)染色、Masson染色，观察其病理结构形态。运用实时(real-time，RT)-PCR、免疫组化(immunohistochemistry)技术检测汉黄芩素对小鼠肺组织中TGF-β1、表皮生长因子(epidermal growth factor，EGF)、波形蛋白、E-钙黏蛋白mRNA和蛋白的表达水平的影响。结果 Biacore垂钓得到的样品经UPLC-Q-TOF-MS鉴定分析为汉黄芩素。汉黄芩素与TGF-β1存在较强的特异性结合。HE染色和Masson染色结果表明，汉黄芩素给药组小鼠肺组织形态结构与对照组相似。免疫组化实验结果表明，与模型组相比，汉黄芩素中、高剂量组TGF-β1、波形蛋白的表达均明显减少(P＜0.05、0.01)，EGF、E-钙黏蛋白的表达升高(P＜0.05、0.01)。RT-PCR实验结果表明，与模型组相比，汉黄芩素各给药组小鼠肺组织TGF-β1、波形蛋白的mRNA表达水平显著降低(P＜0.05、0.001)，EGF和E-钙黏蛋白的mRNA表达显著升高(P＜0.05)。结论 汉黄芩素能够下调TGF-β1和波形蛋白的表达，同时上调EGF和E-钙黏蛋白的表达，从而达到治疗肺炎支原体肺炎及肺组织纤维化的目的。

Objective To investigate the mechanism of wogonin in the treatment of mycoplasma pneumoniae pneumonia by regulating the expression of epithelial-mesenchymal formation factor. Methods UPLC-Q-TOF-MS was used to detect the active ingredient wogonin in Qinbai Qingfei Concentrated Pills (芩百清肺浓缩丸), and it was bound to transforming growth factor-β1 (TGF-β1) by Biacore fishing technique, and the specificity of binding was verified by kinetic analysis. Tripos SYbyl-X2.1.1 and Discovery Studio2016 software was used to perform molecular docking between wogonin and TGF-β1, and the binding mode between drug molecules and receptor proteins was visualized. The lung pathological tissue sections of mice were prepared by animal experiments. Hematoxylin-eosin (HS) staining and Masson staining were performed to observe the pathological structure and morphology of the staining. The effects of wogonin on mRNA and protein expression levels of TGF-β1, epidermal growth factor (EGF), vimentin, E-cadherin in mouse lung tissue by real-time (RT) -PCR and immunohistochemistry method. Results The samples obtained from Biacore fishing were identified and analyzed as wogonin by UPLC-Q-TOF-MS. There was strong specific binding between wogonin and TGF-β1. The results of HE staining and Masson staining showed that the morphological structure of lung tissue in the wogonin group was similar to that of the control group. Immunohistochemical test results showed that compared with model group, the expression levels of TGF-β1 and vimentin in medium and high dose wogonin groups were significantly decreased (P < 0.05, 0.01), and the expressions of EGF and E-cadherin were increased (P < 0.05, 0.01). The results of RT-PCR showed that compared with the model group, the mRNA expression levels of TGF-β1 and vimentin in lung tissue of mice in wogonin administration groups was significantly decreased (P < 0.05, 0.001), while the mRNA expression levels of EGF and E-cadherin was significantly increased (P < 0.05). Conclusion Wogonin can down-regulate the expression of TGF-β1 and vimentin, and up-regulate the expression of EGF and E-cadherin, so as to achieve the purpose of treating mycoplasma pneumoniae pneumonia and lung tissue fibrosis.

国家自然科学基金项目(82174060)；黑龙江省自然科学基金项目(LH2021H074)；黑龙江省卫生健康委科研课题(20210202040011)