基于NF-κB和STAT3探讨白芍总苷对化学性肝损伤肝阴虚证大鼠的作用机制

Mechanism of total glucosides of paeony on chemical liver injury rats with liver yin deficiency syndrome based on NF-κB and STAT3

关键词:  白芍总苷  化学性肝损伤  肝阴虚证  炎症因子  NF-κB

Key words:  total glucosides of paeony  chemical liver injury  liver yin deficiency syndrome  inflammatory factors  NF-κB

摘要:

[目的] 基于核因子-κB（NF-κB）和信号转导和转录激活因子3（STAT3）研究白芍总苷对化学性肝损伤肝阴虚型大鼠的保护作用机制。[方法] 将雄性SD大鼠随机分为空白组、模型组、一贯煎组（6.35 g/kg）、白芍总苷组（50 mg/kg）。采用20%四氯化碳（CCL₄）橄榄油溶液联合30 mg/kg甲状腺片造模，造模的同时给药，时间为6周。观察大鼠体质量、肛温、摄食量、摄水量、舌面干湿度，以及测定血清环磷酸腺苷（cAMP）、环磷酸鸟苷（cGMP）含量、cAMP/cGMP。测定血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、γ-谷氨酰转肽酶（γ-GT）、碱性磷酸酶（ALP）、总胆红素（TBIL）、总胆汁酸（TBA）的含量变化并取肝脏进行Masson染色评估肝纤维化程度；测定血清白细胞介素-1（IL-1）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、白细胞介素-10（IL-10）、白细胞介素-1受体拮抗剂（IL-1Ra）的水平及大鼠肝脏NF-κB p65、STAT3 mRNA及蛋白水平。[结果] 与模型组相比，白芍总苷组体质量上升（P>0.05）；白芍总苷组肛温下降（P<0.01），24 h摄食量和饮水量减少（P<0.01），舌面干湿度增加（P<0.01）；白芍总苷组cAMP降低，cGMP升高，cAMP/cGMP下降（P<0.01）；白芍总苷组ALT、AST、γ-GT、ALP、TBIL均下降（P<0.01），TBA含量无统计意义（P>0.05）；白芍总苷组肝纤维化程度明显减轻；白芍总苷组IL-1、IL-6、TNF-α均下降，IL-10、IL-1Ra水平均上升（P<0.05或P<0.01）；白芍总苷组NF-κB p65 mRNA和蛋白表达显著下降（P<0.01），STAT3 mRNA和蛋白表达升高（P<0.01）。[结论] 研究结果显示，白芍总苷对肝阴虚型肝损伤具有较好的保护作用，可能通过抑制NF-κB的激活，促进STAT3的表达，从而减少促炎细胞因子分泌，增加抗炎细胞因子释放。

Abstract:

[Objective] To study the protective effect and mechanism of total glucosides of paeony(TGP) on chemical liver injury rats with liver yin deficiency type based on nuclear factor-κB(NF-κB) and signal transducer and activator of transcription 3(STAT3). [Methods] Male SD rats were randomly divided into the blank group,the model group,the consistent decoction group(6.35 g/kg),and the TGP group(50 mg/kg). Rats were modelled with 20% carbon tetrachloride(CCL₄) olive oil solution combined with 30 mg/kg thyroid tablets,which were administered at the same time as the modelling for a period of 6 weeks. Body weight,anal temperature,food intake,water intake,dry and wet tongue surface were observed,and the content of serum cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP) and cAMP/cGMP were determined. The serum content of alanine aminotransferase(ALT),aspartate aminotransferase(AST),γ-glutamyl transpeptidase(γ-GT),alkaline phosphatase(ALP),total bilirubin(TBIL),and total bile acid(TBA) were measured and Masson staining was used to evaluate the extent of liver fibrosis. The levels of serum interleukin-1(IL-1),interleukin-6(IL-6),tumor necrosis factor α(TNF-α),interleukin-10(IL-10),interleukin-1 receptor antagonist(IL-1Ra) and the mRNA and protein expressions of NF-κB p65 and STAT3 in rat liver were determined. [Results] Compared with the model group,the body weight of rats in TGP group increased(P>0.05). In TGP group,the anal temperature decreased(P<0.001),24 h food and water intake decreased(P<0.01),and dry and wet tongue surface increased(P<0.01). In TGP group,cAMP decreased,cGMP increased,and cAMP/cGMP decreased(P<0.01). ALT,AST,γ-GT,ALP and TBIL all decreased(P<0.01),while TBA content had no statistical significance(P>0.05). The degree of liver fibrosis in TGP group was significantly reduced. In TGP group,IL-1,IL-6 and TNF-α decreased,while IL-10 and IL-1Ra increased(P<0.05 or P<0.01). The protein and mRNA levels of NF-κB p65 were significantly down regulated in TGP group(P<0.01),while the protein and mRNA levels of STAT3 were significantly elevated(P<0.01). [Conclusion] The result showed that TGP had a good protective effect on hepatic injury of liver yin deficiency,which may reduce the secretion of pro-inflammatory cytokines and increase the release of anti-inflammatory cytokines by inhibiting the activation of NF-κB and promoting the expression of Stat3.