中医学报

目的：观察天龙竭胶囊对博来霉素诱导肺纤维化(pulmonary fibrosis, PF)大鼠的影响，探讨天龙竭胶囊改善PF的作用机理。方法：60只Wistar大鼠随机分为空白组、模型组、天龙竭胶囊低剂量组(0.51 g·kg^-1)、天龙竭胶囊中剂量组(1.01 g·kg^-1)、天龙竭胶囊高剂量组(2.02 g·kg^-1)、吡非尼酮组(0.12 mg·kg^-1)。除空白组外，其余各组大鼠采用气管内滴注博来霉素(5 mg·kg^-1)复制肺纤维化模型。造模14d后，各组大鼠灌胃给予相应药物干预，空白组和模型组灌胃给予等体积生理盐水，每日1次，连续28 d。取大鼠腹主动脉血进行血气分析；采用试剂盒检测肺组织中羟脯氨酸(hydroxyproline, HYP)的含量；HE染色和Masson染色观察大鼠肺组织病理改变；RT-PCR和Western blot检测肺组织α-平滑肌肌动蛋白(alpha-smooth muscle actin, α-SMA)、E-钙黏蛋白(E-Cadherin)、Ⅰ型胶原(collagenⅠ,COL-Ⅰ)及COL-Ⅲ蛋白和mRNA的表达水平。结果：与空白组比较，模型组大鼠肺指数、肺组织HYP水平显著升高(P<0.01),氧分压(partial pressure of oxygen, PO₂)、血氧饱和度(oxygen saturation, SO₂)显著降低(P<0.01),肺组织中COL-Ⅰ、COL-Ⅲ、α-SMA、E-Cadherin mRNA和蛋白表达水平显著上调(P<0.05);与模型组比较，天龙竭胶囊可显著降低大鼠肺指数、肺组织HYP水平(P<0.05),显著升高PO₂、SO₂水平(P<0.05),显著降低肺组织COL-Ⅰ、COL-Ⅲ、α-SMA、E-Cadherin mRNA和蛋白表达水平(P<0.05)。HE染色和Masson染色显示，模型组大鼠肺泡结构破坏，大量炎性细胞浸润，肺间质扩张，肺泡间隔增厚，肺组织胶原沉积明显增多，纤维沉积形成纤维化病灶；天龙竭胶囊可明显改善模型大鼠的肺组织病变，以高剂量效果最为明显。结论：天龙竭胶囊可明显改善PF大鼠病理损伤，改善肺功能及肺纤维化，其作用可能与调节α-SMA/E-Cadherin的表达进而参与上皮-间质转化相关。

Objective: To observe the effect of Tianlongjie Capsule on bleomycin induced pulmonary fibrosis(PF) in ratsα-The impact of SMA/E cadherin and the mechanism by which Tianlongjie Capsule improves PF are explored.Method: Sixty Wistar rats were randomly divided into a blank group, a model group, a low-dose group of Tianlongjie Capsule(0.51 g·kg^-1),a medium dose group of Tianlongjie Capsule(1.01 g·kg^-1),a high-dose group of Tianlongjie Capsule(2.02 g·kg^-1),and a pirfenidone group(0.12 mg·kg^-1).Except for the blank group, rats in all other groups were given tracheal instillation of bleomycin(5 mg·kg^-1) to replicate pulmonary fibrosis models.After 14 days of modeling, each group of rats was given corresponding drug intervention with gavage.The blank group and model group were given equal volume of physiological saline with gavage once a day for 28 consecutive days.Take blood from the abdominal aorta of rats for blood gas analysis.A reagent kit was used to detect the content of hydroxyproline(HYP) in lung tissue.Masson staining and HE staining were used to observe pathological changes in rat lung tissue.RT-PCR and Western blot was used to detect the expression levels of α-Alpha smooth muscle actin, α-SMA,E-cadherin, collagen Ⅰ(COL-Ⅰ),COL-Ⅲ protein and mRNA in the lung tissue.Results: Compared with the blank group, the lung index and lung tissue HYP level of the model group rats were significantly increased(P<0.01),while partial pressure of oxygen(PO₂) and oxygen saturation(SO₂) were significantly reduced(P<0.01).The expression levels of COL-Ⅰ,COL-Ⅲ,and α-SMA as well as E-Cadherin protein and mRNA were significantly upregulated(P<0.05);Compared with the model group, Tianlongjie Capsule can significantly reduce lung index and lung tissue HYP levels in rats(P<0.05),significantly increase PO₂ and SO₂(P<0.05),and significantly decrease the expression levels of lung tissue COL-Ⅰ and COL-Ⅲ α-SMA,E-Cadherin protein and mRNA significantly reduce(P<0.05).HE staining and Masson staining suggested that in the model group of rats, the alveolar structure was damaged, a large number of inflammatory cells infiltrated, the lung interstitium expanded, the alveolar septa thickened and the collagen deposition and fiber deposition in the lung tissue significantly increased, forming fibrotic lesions.Tianlongjie Capsule can significantly improve lung tissue lesions in model rats, with high-dose effects being the most significant.Conclusion: Tianlongjing can significantly improve pathological damage, lung function, and pulmonary fibrosis in PF rats, and its effect may be related to the regulation of the expression of cadherin-αSMA/E and their involvement in epithelial mesenchymal transition.