南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (4): 644-651.doi: 10.12122/j.issn.1673-4254.2024.04.04

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参七虫草方通过ASS1/src/STAT3信号通路改善肺纤维化大鼠的炎症反应

何 程，陈 炜，张念志，栾 军，王三凤，张 尤   

  1. 安徽中医药大学第一附属医院呼吸科，安徽 合肥 230031
  • 发布日期:2024-04-29

Shenqi Chongcao Formula ameliorates inflammatory response in rats with pulmonary fibrosis by activating the ASS1/src/STAT3 signaling pathway

HE Cheng, CHEN Wei, ZHANG Nianzhi, LUAN Jun, WANG Sanfeng, ZHANG You   

  1. Department of Respiratory Medicine, First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China
  • Published:2024-04-29

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摘要： 目的 观察参七虫草方对肺纤维化大鼠精氨酸琥珀酸盐合成酶-1（ASS1）/src酪氨酸激酶（src）/信号转导和转化激活因子3（STAT3）通路相关因子表达的影响，探讨其治疗肺纤维化的作用机制。方法 选择120只SPF级SD雄性大鼠，采用随机数字表法分为5组：空白组、模型组、参七虫草方组、吡非尼酮组、精氨酸脱亚胺酶（ADI）腹腔注射组（24只/组）。除空白组以外，4组均使用一次性气管内滴注博来霉素（BLM）诱导制备特发性肺纤维化的大鼠模型。造模成功后，参七虫草组予参七虫草方汤剂（0.423 g/kg）灌胃；吡非尼酮组予溶于生理盐水的吡非尼酮胶囊粉末（10 mL·kg-1·d-1）灌胃2次；ADI腹腔注射组予以2.25 mg/（kg·d）腹腔注射，1次/3 d；空白组、模型组分别以10 mL/（kg·d）生理盐水灌胃；给药疗程28 d。于第7、14、21、28天分4批处死大鼠，分离肺组织并计算肺指数。采用苏木素-伊红、马松染色观察大鼠肺组织的组织病理学；吉姆萨染色分析BALF中不同种类的炎症细胞；采用酶联免疫吸附测定法检测血清趋化因子配体2（CCL2）、转化生长因子β1（TGF-β1）的含量；蛋白免疫印迹法测定肺组织src、p-srcTry529、STAT3、p-STAT3Try705的蛋白表达；采用实时荧光定量聚合酶链式反应技术检测肺组织ASS1、src、STAT3的表达水平。结果 参七虫草组、吡非尼酮组和ADI腹腔注射组各时间点中性粒细胞、巨噬细胞、淋巴细胞均低于同一时间点的模型组（P<0.05）。在相同时间点，参七虫草组、吡非尼酮组和ADI腹腔注射组大鼠血清CCL2、TGF-β1水平均低于模型组（P<0.05）。参七虫草组、吡非尼酮组和ADI腹腔注射组的p-srcTry529、p-STAT3Try705蛋白表达水平及肺组织ASS1、src、STAT3 mRNA均低于同一时间点的模型组（P<0.05）。结论 参七虫草方可以缓解大鼠的肺纤维化程度，其机制可能通过调控ASS1/src/STAT3信号通路的活化从而缓解BLM大鼠肺纤维化的炎症反应。

关键词: 参七虫草方；精氨酸；特发性肺纤维化；ASS1/src/STAT3信号通路；炎症反应

Abstract: Objective To observe the effect of Shenqi Chongcao (SQCC) Formula on the ASS1/src/STAT3 signaling pathway in a rat model of lung fibrosis and explore its therapeutic mechanism. Methods A total of 120 male SD rats were divided equally into 5 groups, including a blank control group with saline treatment and 4 groups of rat models of idiopathic pulmonary fibrosis induced by intratracheal instillation of bleomycin. One day after modeling, the rat models were treated with daily gavage of 10 mL/kg saline, SQCC decoction (0.423 g/kg), pirfenidone (10 mL/kg), or intraperitoneal injection of arginine deiminase (ADI; 2.25 mg/kg, every 3 days) for 28 days. After the treatments, the lung tissues of the rats were collected for calculating the lung/body weight ratio, observing histopathology using HE and Masson staining, and analyzing the inflammatory cells in BALF using Giemsa staining. Serum chemokine ligand 2 (CCL2) and transforming growth factor- β1 (TGF-β1) levels were measured with ELISA. The protein expressions of src, p-srcTry529, STAT3, and p-STAT3Try705 and the mRNA expressions of ASS1, src and STAT3 in the lung tissues were detected using Western blotting and RT- qPCR. Results The neutrophil, macrophage and lymphocyte counts and serum levels of CCL2 and TGF-β1 were significantly lower in SQCC, pirfenidone and ADI treatment groups than in the model group at each time point of measurement (P<0.05). P-srcTry529 and p-STAT3Try705 protein expression levels and ASS1, src, and STAT3 mRNA in the lung tissues were also significantly lower in the 3 treatment groups than in the model group (P<0.05). Conclusion SQCC Formula can alleviate lung fibrosis in rats possibly by activating the ASS1/src/STAT3 signaling pathway in the lung tissues.

Key words: Shenqi Chongcao Formula; arginine; idiopathic pulmonary fibrosis; ASS1/src/STAT3 signaling pathway; inflammatory response