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/3d;空白组、模型组分别以10mL/(kg·d)生理盐水灌胃;给药疗程28d。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-src^(Try529)、STAT3、p-STAT3^(Try705)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-src^(Try529)、p-STAT3^(Try705)蛋白表达水平及肺组织ASS1、src、STAT3mRNA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。
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^(Try705)(TGF-β1)levels were measured with ELISA.The protein expressions of src,p-src^(Try529),STAT3,and p-STAT3 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,Try529 pirfenidone and ADI treatment groups than in the model group at each time point of measurement(P<0.05).P-src and p-^(Try705)STAT3 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.
作者
何程
陈炜
张念志
栾军
王三凤
张尤
HE Cheng;CHEN Wei;ZHANG Nianzhi;LUAN Jun;WANG Sanfeng;ZHANG You(Department of Respiratory Medicine,First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2024年第4期644-651,共8页
Journal of Southern Medical University
基金
国家自然科学基金青年基金(81704060)
国家中医药管理局青年岐黄学者人才支持项目(国中医药人教发[2020]7号文)
安徽省临床医学研究转化专项(202304295107020106)。
