化浊解毒调肝方对乙型肝炎大鼠肝纤维化、免疫功能及Ras/ERK信号通路的影响

Study of Huozhujiedu Tiaogan prescription on liver fibrosis,immune function and Ras/ERK signaling pathway in rats with hepatitis B

目的:研究化浊解毒调肝方对乙型肝炎大鼠肝纤维化、免疫功能及Ras/ERK信号通路的影响。方法:将50只大鼠随机分为正常组、模型组、拉米夫定阳性对照组、化浊解毒调肝方低剂量实验组、化浊解毒调肝方高剂量组,每组10只。除正常组外均建立AAV-HBV大鼠模型,建模成功后,阳性对照组予以0.3 mg/kg拉米夫定灌胃,低、高剂量实验组分别以生药量1.5 g/ml、3 g/ml化浊解毒调肝方剂灌胃,其余建模组均以同剂量生理盐水灌胃,5组大鼠均持续治疗28 d。治疗结束后,取5组大鼠血清及肝组织,用ELISA法分别检测5组大鼠血清中GPT、GOT、IFN-γ、IL-2、IL-4、HA、LN、PCⅢ含量,用免疫印记检测肝组织中Ras、p-Erk/Erk蛋白水平,HE染色观察肝组织病理形态。结果:与正常组相比,模型组大鼠血清中GPT、GOT、IL-4、HA、LN、PCⅢ含量均升高,IFN-γ、IL-2水平降低,肝组织中Ras、p-Erk水平明显升高(P<0.05);与模型组相比,阳性对照组、低/高剂量实验组GPT、GOT、IFN-γ、IL-2、IL-4、HA、LN、PCⅢ、Ras、p-Erk蛋白水平均有明显改善(P<0.05),阳性对照组与低剂量实验组大鼠之间比较,各指标水平无明显差异(P>0.05),与其他建模组相比,高剂量实验组各指标水平均有明显改善(P<0.05)。结论:化浊解毒调肝方可有效降低乙型肝炎大鼠肝纤维化程度,提高免疫功能,机制与调控Ras/ERK信号通路相关,并呈现剂量依赖性。

Objective:To study the effects of Huazujiedu Tiogan Prescription on liver fibrosis,immune function,and Ras/ERK signaling pathway in rats with hepatitis B.Methods:A total of 50 rats was randomly divided into the normal group,model group,positive control group,low-dose experimental group,and high-dose experimental group with 10 rats.The AAV-HBV rat models were established in all groups except the normal group.After successful modeling,the positive control group was given 0.3 mg/kg lamivudine by intragastric administration.The low-dose and high-dose experimental groups were given intragastric administration of turbidity-detoxification and liver-regulation prescription with a dose of 1.5 g/ml and 3 g/ml,respectively.And the other modeling groups were given the same dose of normal saline.All five groups were treated continuously for 28 days.After treatment,the serum and liver tissues of the five groups of rats were taken,and the contents of GPT,GOT,IFN-γ,IL-2,IL-4,HA,LN,and PCⅢin the serum of the five groups of rats were detected by Elisa.The levels of Ras and p-Erk/Erk protein in the liver tissue were detected by immunoimprinting,and the pathological morphology of the liver tissue was observed by HE staining.Results:Compared with the normal group,the contents of GPT,GOT,IL-4,HA,LN,and PCⅢin the serum of the model group were increased,the levels of IFN-γand IL-2 were decreased,and the levels of Ras and p-ERK in liver tissue were significantly increased(P<0.05).Compared with the model group,the protein levels of GPT,GOT,IFN-γ,IL-2,IL-4,HA,LN,PCⅢand Ras/p-ERK in the positive control group and low/high dose experimental group were significantly improved(P<0.05).There was no significant difference in the level of each index(P>0.05),and compared with other modeling groups,the level of each index in the high-dose experimental group was significantly improved(P<0.05).Conclusion:Reducing turbidity,detoxifying,and regulating the liver can effectively reduce the degree of liver fibrosis and improve the immune function of hepatitis B rats.The mechanism is related to the regulation of the Ras/ERK signaling pathway,and it is dose-dependent.