茵芪肝复颗粒对胆汁淤积性肝炎大鼠的改善作用及对相关细胞和炎症因子的影响

Yinzhi Ganfu granules for treatment of chronic cholestatic hepatitis in rats: Efficacy and influence on cytokines and chemokines

背景茵芪肝复颗粒具有疏肝补脾、清热解毒利湿功效,但其对肝功能影响及作用机制尚不明确.本研究经过分析茵芪肝复颗粒对胆汁淤积性肝炎(cholestatic hepatitis,CP)大鼠肝功能影响,为临床治疗提供一些实验室依据.目的探讨茵芪肝复颗粒对大鼠CP的改善作用及对血清炎症因子、氧化应激指标及法尼酯X受体(farnesyl Xreceptor, FXR)的影响.方法 27只雄性Wistar大鼠根据处理方法不同分为对照组(n=9,健康大鼠,不予任何处理)、模型组[n=9,以α-异硫氰酸萘酯(α-naphthyl isothiocyanate, ANIT)灌胃法复制CP大鼠模型]、茵芪肝复颗粒组(n=9,ANIT灌胃法复制CP大鼠模型后给予茵芪肝复颗粒处理),比较3组肝组织病理变化、肝功能指标[谷草转氨酶(aspartate aminotransferase, AST)、谷丙转氨酶(alanineaminotransferase, ALT)、碱性磷酸酶(alkaline phosphatase, ALP)、总胆红素(totalbilirubin,TB)、谷氨酰转肽酶(glutamyltranspeptidase, GGT)]、血清细胞因子水平[IL-17、IL-4、干扰素-γ(Interferon-γ,I F N-γ)、超氧化物歧化酶(s u p e r o x i d ed i s m u t a s e,SOD)、丙二醛(malondialdehyde,MDA)]、肝组织IL-17、IL-4、IFN-γ、SOD、MDA、FXRm RNA水平,采用LOGISTIC多元回归方程分析影响肝功能因素.结果肝组织病理:对照组肝脏细胞、结构无异常;模型组肝脏细胞、结构存在明显异常;茵芪肝复颗粒组肝脏小叶结构完整清晰,肝细胞肿胀,肝索呈放射状;茵芪肝复颗粒组AST、ALT、ALP、TB、GGT低于模型组,高于对照组(P<0.05);茵芪肝复颗粒组IL-17、IFN-γ、MDA高于对照组,SOD低于对照组(P <0.05);模型组IL-17、IFN-γm RNA高于对照组,IL-4、SOD、FXRm RNA低于对照组(P <0.05);茵芪肝复颗粒组IL-17、IFN-γm RNA低于模型组, IL-4、SOD、FXRm RNA高于模型组(P <0.05);茵芪肝复颗粒组IL-17、IFN-γm RNA高于对照组, SOD、FXR m RNA低于对照组(P <0.05);IL-17、IFN-γ、MDA与CP呈正相关, SOD、FXR与CP呈负相关(P <0.05).结论茵芪肝复颗粒能明显改善CP大鼠肝功能、肝细胞结构形态,其机制可能与调控IL-17、IFN-γ、MDA等因子,缓解炎症反应、提高抗氧化能力等有关.

BACKGROUND Yinzhi Ganfu granules has the functions of soothing the liver and replenishing the spleen, clearing away heat, detoxifying, and removing dampness, but its effect on liver function and its mechanism of action are still unclear. This study analyzed the efficacy of Yinzhi Ganfu granules in the treatment of cholestatic hepatitis(CP) in rats to provide an experimental basis for clinical treatment of this disease.AIM To investigate the efficacy of Yinzhi Ganfu granules in the treatment of cholestasis hepatitis(CP) in rats and its effect on serum inflammatory factors, oxidative stress index, and farnesyl X receptor(FXR).METHODS Twenty-seven male Wistar rats were divided into a control group(n = 9, healthy rats, given no treatment), model group [n = 9, treated with α-naphthyl isothiocyanate(ANIT) to replicate a CP model], and Yinzhi Ganfu granules group(n = 9, CP rats administered with Yinzhi Ganfu granules). Pathological changes, liver function indexes [aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP), total bilirubin(TB), and glutamyl transpeptidase(GGT)], and serum cytokine levels [interleukin(IL)-17, IL-4, and interferon-γ(IFN-γ)] were assayed. Superoxide dismutase(SOD), malondialdehyde(MDA), liver tissue IL-17, IL-4, IFN-γ, SOD, MDA, and FXR m RNA levels were also analyzed. By using multiple logistic regression, the factors that may affect liver function were evaluated.RESULTS There were no abnormalities in liver cells and structure in the control group;liver cells and structures in the model group were abnormal;the liver lobule structure in the Yinzhi Ganfu granules group was intact and clear, hepatocytes were swollen, and hepatic cord was radial. AST, ALT, ALP, TB, and GGT in the Yinzhi Ganfu granules group were significantly lower than those of the model group, but significantly higher than those of the control group(P < 0.05). IL-17, IFN-γ, and MDA were significantly higher and SOD was significantly lower in the Yinzhi Ganfu granules group than in the control group(P < 0.05);IL-17 and IFN-γ m RNAs were significantly higher and IL-4, SOD, and FXR m RNAs were significantly lower in the model group than in the control group(P < 0.05). IL-17 and IFN-γ m RNAs were significantly lower and IL-4, SOD, and FXR m RNAs were significantly higher in the Yinzhi Ganfu granules group than in the model group(P < 0.05). IL-17 and IFN-γ m RNAs in the Yinzhi Ganfu granules group were significantly higher than those of the control group, and SOD and FXR m RNAs were significantly lower than those of the control group(P < 0.05);IL-17, IFN-γ, and MDA were positively correlated with CP, while SOD and FXR were negatively correlated with CP(P < 0.05).CONCLUSION Yinzhi Ganfu granules can significantly improve liver function and liver cell structure and morphology of CP rats via mechanisms possibly related to alleviating the inflammatory reaction and improving antioxidant capacity by regulating IL-17, IFN-γ, MDA, and other cytokines.