双黄连口服液对3型鼠肝炎病毒致急性肝衰竭模型C57BL/6J裸小鼠的保护作用机制研究

Study on the mechanism of protective effect of Shuanghuanglian oral liquid on C57BL/6J nude mice with acute liver failure induced by hepatitis virus type 3

目的观察双黄连口服液对3型鼠肝炎病毒(MHV-3)致裸小鼠急性肝衰竭(ALF)模型的影响,探讨其可能作用机制。方法将32只C57BL/6J裸小鼠随机分为正常组、急性肝衰竭组、双黄连低剂量组和双黄连高剂量组,每组8只。除正常组外,其余组均采用腹腔注射MHV-3建立急性肝衰竭模型。建模成功后,双黄连低剂量组和双黄连高剂量组分别给予0.25 mL/(10 g·d)和0.5 mL/(10 g·d)双黄连口服液灌胃,正常组、急性肝衰竭组分别给予等体积生理盐水灌胃,均1次/d,连续灌胃28 d。取静脉血检测天门冬氨酸氨基转移酶(AST)、谷氨酰转移酶(GGT)、总胆红素(TBil)、丙氨酸氨基转移酶(ALT)、胆汁酸(TBA)水平,取肝脏组织检测干扰素-γ(IFN-γ)、转化生长因子-β1(TGF-β1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-9(IL-9)表达水平。结果急性肝衰竭组小鼠血清AST、GGT、TBil、ALT、TBA水平和肝组织中IFN-γ、TNF-α、TGF-β1、IL-1β、IL-6、IL-9表达水平均显著高于正常组(P均<0.05);双黄连高剂量组和双黄连低剂量组小鼠血清AST、GGT、TBil、ALT、TBA水平和肝组织中IFN-γ、TNF-α、TGF-β1、IL-1β、IL-6、IL-9表达水平均显著低于急性肝衰竭组(P均<0.05),且双黄连高剂量组各指标水平均显著低于双黄连低剂量组(P均<0.05)。结论双黄连口服液对MHV-3致C57BL/6J裸小鼠急性肝衰竭具有保护作用,主要体现在改善肝功能并抑制细胞因子合成、减轻炎症性肝损伤方面。

Objective It is to observe the effect of Shuanghuanglian oral liquid on acute liver failure(ALF)model induced by murine hepatitis virus strain 3(MHV-3)in nude mice and analyze its possible protective mechanism.Methods 32 C57BL/6J nude mice were randomly divided into four groups:normal group,acute liver failure group,Shuanghuanglian low dose group and Shuanghuanglian high dose group,8 mice in each group.All the groups except the normal group were injected with MHV-3 intraperitoneally to establish the acute liver failure model.After successful modeling,the Shuanghuanglian low-dose group and Shuanghuanglian high-dose group were given 0.25 mL/(10g·d)and 0.5 mL/(10g·d)Shuanghuanglian oral solution,respectively,the rats in the normal group and the acute liver failure group were given the same volume of saline,once a day for 28 days.The venous blood was taken to detect the levels of aspartate aminotransferase(AST),glutamyltransferase(GGT),total bilirubin(TBil),alanine aminotransferase(ALT),and bile acid(TBA),liver tissue was taken to detect the levels of expression of interferon-γ(IFN-γ),transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-9(IL-9).Results The levels of AST,GGT,TBil,ALT,TBA in serum and IFN-γ,TNF-α,TGF-β1,IL-1β,IL-6,IL-9 in liver tissues of mice with acute liver failure were significantly higher than those in the normal group(P<0.05);the levels of AST,GGT,TBil,ALT,TBA in serum and IFN-γ,TNF-α,TGF-β1,IL-1β,IL-6 and IL-9 in liver tissues of mice in Shuanghuanglian high dose group and Shuanghuanglian low dose group were significantly lower than those in the acute liver failure group(all P<0.05),and the levels of all indicators in the Shuanghuanglian high-dose group were significantly lower than those in the Shuanghuanglian low-dose group(all P<0.05).Conclusion Shuanghuanglian oral liquid has protective effect on C57BL/6J nude mice with acute liver failure induced by MHV-3.This effect is mainly manifested in improving liver function,inhibiting cytokine synthesis and reducing inflammatory liver injury.