补气活血通络方干预肺纤维化大鼠PI3K/AKT信号通路研究

Intervention to PI3K/AKT signaling pathway by Buqi Huoxue Tongluo Fang in rats with idiopathic pulmonary fibrosis

目的探讨补气活血通络方对肺间质纤维化模型大鼠磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/AKT)信号通路的影响。方法将大鼠随机分为正常组,模型组,泼尼松组,补气活血通络方低、中、高剂量组,共6组,除正常组外,其余各组以博来霉素3 mg/kg造模,造模后第2天补气活血通络方低、中、高,组分别给予补气活血通络方水煎剂8. 25、16. 5、33 g/kg灌胃,泼尼松组给予醋酸泼尼松片混悬液3. 6 mg/kg灌胃,正常组和模型组给予等量生理盐水灌胃。于第14天及第28天分别2个时间点,观察大鼠一般状况的改变,以免疫组化法观察第10号染色体同源丢失性磷酸酶-张力蛋白(PTEN)、PI3K、AKT、雷帕霉素靶蛋白(mTOR)蛋白表达的差异,以RT-PCR法观察基因表达的差异。结果与模型组及泼尼松组比较,补气活血通络方能改善大鼠一般状况及体重(P <0. 05),上调PTEN并下调PI3K、AKT、mTOR的蛋白和基因表达(P <0. 05)。结论补气活血通络方通过干预PI3K/AKT信号通路的活化,从而起到对抗纤维化的作用。

Objective To investigate the influence of Buqi Huoxue Tongluo Fang( Qi-tonifying Bloodactivating Collateral-freeing Formula,BHTF) on PI3K/AKT signaling pathway in rat model of idiopathic pulmonary fibrosis( IPF). Methods All rats were randomly divided into 6 groups including normal group,model group,prednison group,low-dose,mid-lose and high-dose BHTF groups. Except of normal group,other groups were given bleomycin( 3 mg/kg) for modeling. After 2 d,all BHTF groups were orally given BHTF decoction in different doses( 8. 25 g/kg,16. 5 g/kg and 33 g/kg),prednison group was orally given prednisone acetate suspension( 3. 6 mg/kg),and normal group and model group were orally given equivalent normal saline. The changes of rat general status were observed,difference in protein expressions of PTEN, PI3K, AKT and mTOR signaling pathway were detected by using immunohistochemistry technique,and difference in genetic expressions was observed by using RT-PCR at2 time points( 14 d and 28 d). Results Compared with model group and prednison group,the rat general status and weight were improved( P < 0. 05),PTEN expression was up-regulated and expressions of PI3K,AKT and mTOR were down-regulated in all BHTF groups( P < 0. 05). Conclusion BHTF can play an anti-fibrosis role through intervening the activation of PI3K/AKT signaling pathway.