蒙药阿嘎如-25对小鼠肺纤维化的影响

Effect of Mongolian medicine Agaru-25 on pulmonary fibrosis in mice

目的蒙药阿嘎如-25对小鼠肺纤维化的影响及其作用机制。方法选取健康SPF级C57BL/6小鼠72只,随机选取10只为正常组,其余62只小鼠麻醉后气管内注入博来霉素建立肺纤维化模型。造模第28天,死亡7只。造模成功后根据随机数字表法将小鼠分为模型组(10只)、西药组(9只)、蒙高组(9只,13 mg/ml)、蒙中组(9只,6.5 mg/ml)、蒙低组(9只)、中药组(9只,3.5 mg/ml),各组小鼠灌胃8周后处死取材。免疫组织化学染色及Masson染色观察小鼠肺组织病理改变及纤维化,Western blot法检测肺组织中转化生长因子-β1(TGF-β1)、受体磷酸酯细胞质蛋白(p-Smad3)、纤维连接蛋白(FN)、骨桥蛋白(OPN)的表达,Tunel法检测细胞凋亡率。结果蒙高组、蒙中组、蒙低组肺组织病理形态均较模型组明显改善;模型组胶原纤维面积百分比高于正常组,各蒙药组胶原纤维面积百分比低于模型组,蒙高组胶原纤维面积百分比低于其他给药组,差异均有统计学意义(P<0.05)。蒙药组TGF-β1、p-Smad3、FN、OPN蛋白表达低于模型组,蒙高组TGF-β1、p-Smad3、FN、OPN蛋白表达低于其他蒙药组,且蒙中组低于蒙低组,差异有统计学意义(P<0.05);各蒙药组细胞凋亡率低于模型组,蒙高组细胞凋亡率低于其他给药组,且蒙中组低于蒙低组,差异有统计学意义(P<0.05)。结论蒙药阿嘎如-25抗肺纤维化的作用机制可能与抑制TGF-β1/Smad3信号通路有关。

Objective The effect of Mongolian medicine Agaru-25 on pulmonary fibrosis in mice.Methods Seventy-two healthy SPF C57BL/6 mice were selected,ten mice were randomly selected as normal group,Bleomycin was injected into the trachea of the other 62 mice after anesthesia to establish pulmonary fibrosis model.On the 28th day of modeling,seven mice died.After successful molding,the mice were divided into model group(n=10),Western medicine group(n=9,13 mg/ml),Mongolian high group(n=9,6.5 mg/ml),Mongolian middle group(n=9,3.25 mg/ml),Mongolian low group(n=9),and Chinese medicine group(n=9)according to random number table method,after eight weeks of intragastric administration,mice in each group were sacrificed for sampling.Immunohistochemical staining and Masson staining were used to observe the pathological changes and fibrosis of lung tissues,Western blot was used to detect the expression of transforming growth factor-β1(TGF-β1),receptor phosphate cytoplasmic protein(p-Smad3),fibronectin(FN)and,osteopontin(OPN)in lung tissues,and Tunel was used to detect the apoptosis rate.Results Compared with model group,the pathological morphology of lung tissue in Mongolian high group,Mongolian middle group and Mongolian low group was significantly improved,the percentage of collagen fiber area in model group was higher than that in normal group,the percentage of collagen fiber area in each administration group was lower than that in model group,and the percentage of collagen fiber area in Mongolian group was lower than that in other administration groups,and the differences were statistically significant(P<0.05).The protein expressions of TGF-β1,p-Smad3,FN,and OPN in the treatment group were lower than those in the model group,and the protein expressions of TGF-β1,p-Smad3,FN,and OPN in the Mongolian high group were lower than those in the other Mongolian medicine groups,and Mongolian middle group were lower than Mongolian low group,and the differences were statistically significant(P<0.05).The apoptosis rate of the each Mongolian medicine groups was lower than that of the model group,and the apoptosis rate of Mongolians group was lower than that of other Mongolian medicine groups,and Mongolian middle group were lower than Mongolian low group,and the differences were statistically significant(P<0.05).Conclusion The anti-pulmonary fibrosis mechanism of the Mongolian medicine Agaru-25 may be related to the inhibition of the TGF-β/Smad signaling pathway.