保金尘肺方对矽肺大鼠肺纤维化及肺组织Akt/Gsk-3β通路的影响

Effects of Baojin Chenfei Formula on fibrosis and lung tissue Akt/Gsk-3βpathway in silicosis rats

目的:探讨保金尘肺方对矽肺大鼠纤维化的干预效果及对蛋白激酶B(Akt)/糖原合酶激酶-3β(Gsk-3β)通路的影响。方法:将48只SD大鼠随机分为空白组、模型组、汉方己甲素组和保金尘肺组,每组12只。采用一次性气管内注入三氧化硅(SiO2)混悬液方法制备矽肺大鼠模型,于造模后第15—28天进行相应的药物灌胃治疗。治疗结束后测定大鼠肺功能,包括肺活量(VC)、潮气量(TV)、呼吸暂停(PAU)和用力最大吸气流速(PIF),观察肺组织病理形态并进行纤维化评分,检测肺组织Col-Ⅰ、Col-Ⅲ表达,羟脯氨酸(HYP)含量,N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)以及Akt/Gsk-3β通路相关mRNA、蛋白水平。结果:与空白组比较,模型组大鼠肺组织结构破坏严重且可见纤维结节,PAU显著延长(P<0.01),纤维化评分、肺组织HYP含量、Col-Ⅰ和Col-Ⅲ蛋白表达、α-SMA mRNA、转化生长因子-β1(TGF-β1)mRNA、N-cadherin蛋白、p-Akt蛋白、p-Gsk-3β蛋白表达均显著升高(P<0.01,P<0.05),VC、TV、PIF、E-cadherin蛋白及mRNA均显著下降(P<0.01,P<0.05)。与模型组比较,保金尘肺组和汉防己甲素组大鼠肺组织纤维化评分、HYP含量、Col-Ⅰ表达、N-cadherin蛋白、p-Gsk-3β蛋白均显著下降(P<0.01,P<0.05),E-cadherin蛋白表达显著升高(P<0.01),保金尘肺组Col-Ⅲ表达、PAU、α-SMA和TGF-β1 mRNA、p-Akt蛋白降低(P<0.05,P<0.01)。与汉防己甲素组比较,保金尘肺组TV、E-cadherin蛋白表达显著升高(P<0.05,P<0.01),Col-Ⅲ表达显著降低(P<0.05)。结论:保金尘肺方可通过抑制上皮-间质转化改善矽肺大鼠纤维化,其机制可能与调控Akt/Gsk-3β信号通路有关。

Objective:To explore the effects of Akt/Gsk-3βpathway by Baojin Chenfei Formula(BJCF)in delaying fibrosis in silicotic rats.Methods:A total of 48 rats were randomly divided into normal group,model group,tetrandrine group and BJCF group,with 12 rats in each group.The silicosis rat model was established by one-time injection of SiO2 through trachea except for the normal group,the rats in each group were treated with the corresponding drugs on the day 15-28 after modeling,the rats were sacrificed and the samples were taken after treatment.After treatment,pulmonary function was measured,including VC,TV,PAU and PIF,the pathological morphology of lung tissue and score fibrosis were observed,Col-I,Col-III,hydroxyproline(HYP),N-cadherin,E-cadherin and Akt/Gsk-3βpathway related mRNA and protein level in lung tissue were measured.Results:Compared with the normal group,the lung tissue structure was severely damaged and fibrous nodules were visible in model group,the PAU were significantly extended(P<0.01),the fibrosis score,the content of HYP,the mRNA relative expression ofα-SMA,TGF-β1,the protein relative expression of Col-I,Col-III,N-cadherin,p-Akt,p-Gsk-3βwere significantly increased(P<0.01,P<0.05);the VC,TV and PIF and the mRNA,protein relative expression of E-cadherin were significantly decreased in the model group(P<0.01,P<0.05).Compared with the model group,the fibrosis score,the content of HYP,Col-I expression,and the protein relative expression of N-cadherin,p-Gsk-3 were significantly decreased in the BJCF group and the tetrandrine group(P<0.01,P<0.05),the protein relative expression of E-cadherin were significantly increased in the BJCF group and the tetrandrine group(P<0.01),the Col-III expression,PAU,α-SMA mRNA,TGF-β1 mRNA and the protein relative expression of p-Akt were significantly decreased in the BJCF group(P<0.05,P<0.01).Compared with tetrandrine group,the TV and the protein relative expression of E-cadherin were significantly increased in BJCF group(P<0.05,P<0.01),the Col-III expression was significantly decreased in BJCF group(P<0.05).Conclusion:BJCF can improve fibrosis in silicotic rats by inhibiting epithelial-mesenchymal transition,and its mechanism may be related to the regulation of Akt/Gsk-3βpathway.