相思藤总黄酮基于FAK/PI3K/Akt信号通路抗CCl4大鼠肝纤维化作用机制的研究

Study on the anti-CCL4 rat liver fibrosis mechanism of Xiangsiteng total flavonoid based on FAK/PI3K/Akt signaling pathway

目的:研究相思藤总黄酮(XSTF)对四氯化碳(CCl4)诱导大鼠HF的保护作用及其机制。方法:雄性SD大鼠80只,随机分为正常对照组、模型组、秋水仙碱组(0.2 mg/kg),XSTF高、中、低剂量组(800 mg/kg、400 mg/kg、200 mg/kg),每组12只,除正常对照组外,其余各组大鼠灌胃给予剂量50%CCl4花生油混合液(1 mL/kg)制备肝纤维化(HF)模型,2次/周。确定大鼠形成HF后连续4周,每日给予大鼠秋水仙碱和XSTF相应剂量药物,正常对照组和模型组给予相应体积的生理盐水。给药期间,除正常对照组外,其余各组同时继续造模处理。末次给药24 h后,收集血清及肝组织,检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;制备肝脏匀浆,检测肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-px)含量;实时荧光定量PCR(qPCR)检测肝组织FAK、PI3K、Akt mRNA表达情况;免疫组化法检测肝组织中PI3K、Akt蛋白表达水平。结果:XSTF干预后可明显抑制CCl4诱导的肝脏氧化应激反应,与模型组大鼠相比,XSTF给药组血清中TNF-α、IL-6含量显著降低(P<0.05或P<0.01);肝组织中MDA含量降低(P<0.05或P<0.01),SOD、GSH-px的活性升高(P<0.05或P<0.01);免疫组化结果显示,PI3K、Akt蛋白表达水平明显上升(P<0.01);qPCR结果显示,FAK、PI3K、Akt的mRNA表达水平明显提升(P<0.01)。结论:XSTF对CCl4诱导的大鼠HF具有明显改善作用,可以明显减小肝组织损伤,其保护机制可能与减轻炎症反应,抑制FAK/PI3K/Akt信号传导通路有关。

Objective:To investigate the protective effect and its mechanism of Xiangsiteng total flavonoid(XSTF)against liver fibrosis induced by CCl4 in rats.Methods:80 male SD rats were randomly divided into normal group,model group,colchicine group(0.2 mg/kg),XSTF high,medium and low dose groups(800,400,200 mg/kg),12 rats in each group.Except for the normal control group,other groups rats were given 50%CCl4 peanut oil mixture(1 mL/kg)by gavage-twice a week to produce the liver fibrosis model.After the formation of hepatic fibrosis(HF)in rats,the corresponding doses of colchicine and XSTF were given to rats by gavage every day for four weeks,the normal group and the model group rats were given the same amount of normal saline.During the administration period,all groups except the normal control group continued the modeling treatment at the same time.After the last administration for 24 h,serum and liver tissues were collected.Rat serum TNF-α and IL-6 levels were detected.The hepatic homogenation was carried out and the levels of Hyp,SOD,MDA and GSH-px in hepatic tissues were tested.Real timefluorescence quantitative PCR was used for detecting the liver tissue FAK,PI3K,Akt mRNA levels.The immunological measures was conducted to detect PI3K and Akt protein levels.Results:After XSTF intervention,liver oxidative stress induced by CCl4 was significantly inhibited.The serum levels of TNF-αand IL-6 in the XSTF group were significantly reduced compared with those in the model group(P<0.05 or P<0.01).The content of MDA in liver tissues was decreased(P<0.05 or P<0.01),while the activities of SOD and GSH-PX were increased(P<0.05 or P<0.01).Immunohistochemical results showed that PI3K and Akt protein expression levels were significantly increased(P<0.01).qPCR results showed that mRNA expression levels of FAK,PI3K and Akt were significantly increased(P<0.01).Conclusion:XSTF has a significant improvement on CCl4-induced HF in rats,which can significantly reduce liver tissue damage,and its protective mechanism may be related to reducing inflammatory response and inhibiting the FAK/PI3K/Akt signaling pathway.