益肺通络方对肺纤维化大鼠上皮间质转化的影响

Effect of Yifei Tongluo Formula on Epithelial-Mesenchymal Transition in Pulmonary Fibrosis Rats

目的:探讨益肺通络方干预大鼠肺纤维化的作用机制。方法:56只SD雄性大鼠,随机选取10只大鼠作为空白组(Control组),剩余56只大鼠通过气管内灌注硫酸博来霉素(5 mg/mL)的方法建立肺纤维化动物模型。造模成功后将存活的40只大鼠随机分为模型组(Model组)、醋酸泼尼松组(PRED组)、益肺通络方低剂量组(YFTL-L组)、益肺通络方高剂量组(YFTL-H组)。PRED组灌胃1.17 mg/kg醋酸泼尼松片,YFTL-L组、YFTL-H组分别灌胃1.027、2.054 g/mL益肺通络方水煎液,Control组和Model组均灌胃等体积生理盐水,连续给药21 d。治疗结束后计算肺脏系数;HE、Masson染色观察肺组织破坏及胶原沉积情况;ELISA检测血清中羟脯氨酸(Hyp)、TNF-α水平;免疫荧光检测肺组织TGF-β1、Col-I、E-cadherin蛋白水平;免疫组化检测肺组织α-SAM、vimentin蛋白水平。结果:与Model组比较,YFTL-L组、YFTL-H组肺组织结构破坏情况均有改善,胶原沉积减少,大鼠肺脏系数和纤维化评分均降低(P<0.01),血清中Hyp、TNF-α水平降低(P<0.01),肺组织中TGF-β1、Col-I、α-SAM、vimentin蛋白表达下调(P<0.01,P<0.05),E-cadherin蛋白水平升高(P<0.05,P<0.01),且随着灌胃药物浓度升高呈现出一定的剂量依赖效应。结论:益肺通络方可延缓大鼠肺纤维化进展,其作用机制可能是通过缓解肺泡炎症、抑制上皮间质转化机制转化发挥抗纤维化作用。

Objective:To explore the mechanism of Yifei Tongluo Formula intervention in pulmonary fibrosis(PF)in rats.Methods:Fifty-six male SD rats were randomly divided into 10 rats for the blank group(control group),and the remaining 56 rats were intratracheally instilled with bleomycin sulfate(5 mg/mL)to establish a PF animal model.After successful modeling,the surviving 40 rats were randomly divided into the model group,prednisone acetate group(PRED group),low-dose Yifei Tongluo Formula group(YFTL-L group),and highdose Yifei Tongluo Formula group(YFTL-H group).The PRED group was gavaged with 1.17 mg/kg prednisone acetate tablets,and the YFTL-L group and YFTL-H group were gavaged with 1.027 and 2.054 g/mL Yifei Tongluo Formula decoction,respectively.The control group and model group were gavaged with an equal volume of physiological saline.The drugs were administered continuously for 21 days.After treatment,the lung coefficient was calculated;HE and Masson staining were used to observe lung tissue damage and collagen deposition;ELISA was used to detect serum hydroxyproline(Hyp)and TNF-αlevels;immunofluorescence was used to detect the protein levels of TGF-β1,Col-I,and E-cadherin in lung tissue;immunohistochemistry was used to detect the protein levels ofα-SMA and vimentin in lung tissue.Results:Compared with the model group,the lung tissue structure damage in the YFTL-L group and YFTL-H group was improved,collagen deposition was reduced,the lung coefficient and fibrosis score decreased(P<0.01),serum levels of Hyp and TNF-αdecreased(P<0.01),the protein expression of TGF-β1,Col-I,α-SMA,and vimentin in lung tissue was downregulated(P<0.01,P<0.05),and E-cadherin protein levels increased(P<0.05,P<0.01).Moreover,there was a certain dose-dependent effect with increasing drug concentration.Conclusion:Yifei Tongluo Formula can delay the progression of PF in rats,and its mechanism of action may be through alleviating pulmonary inflammation and inhibiting epithelial-mesenchymal transition to exert anti-fibrotic effects.