温阳解毒化瘀方对慢加急性肝衰竭大鼠肝组织白细胞介素-10/信号转导因子和转录激活因子3及相关因子表达的影响

Efficacy of the Wenyang Jiedu Huayu prescription on the expression of IL-10/STAT3 and related factors in liver of acute-on-chronic liver failure rats

目的:探讨温阳解毒化瘀方对慢加急性肝衰竭大鼠肝组织白细胞介素(Interleukin,IL)-10/信号转导因子和转录激活因子3(Signal Transducer and Activator of Transcription 3,STAT3)表达及其相关炎症因子分泌的调控作用。方法:将健康SD雄性大鼠随机分为正常组、模型组、中药组,除正常组外,其余均造模。采用皮下多点注射牛血清白蛋白致敏联合D-氨基半乳糖胺(D-Gal)/脂多糖(LPS)腹腔急性攻击法构建慢加急性肝衰竭大鼠模型,正常组以等剂量生理盐水腹腔注射。中药组给予温阳解毒化瘀方灌胃,正常组和模型组给予等剂量蒸馏水灌胃。检测各组大鼠急性攻击后1 h、12 h、24 h肝功能、肝组织病理形态学改变、炎症因子[肿瘤坏死因子(Tumor Necrosis Factor,TNF)-α、IL-1β、转化生长因子(Transforming Growth Factor,TGF)-β、IL-10]、STAT3相关蛋白、基因表达。结果:①肝功能:与正常组比较,其余各组在各时间点谷丙转氨酶、谷草转氨酶含量均明显升高(P<0.01);与模型组比较,中药组在各时间点谷丙转氨酶、谷草转氨酶含量均明显下降(P<0.01)。②炎症因子水平:与正常组比较,其余各组炎症因子(TNF-α、IL-1β、TGF-β、IL-10)各时间点表达均显著升高(P<0.01);与模型组比较,中药组在各时间点促炎因子(TNF-α、IL-1β)表达均显著下降(P<0.01),抑炎因子(TGF-β、IL-10)表达均显著上升(P<0.01)。③肝组织病理评分:与正常组比较,其余各组各时间点肝损伤均显著加重(P<0.01);与模型组比较,中药组肝损伤减轻(P<0.05)。④IL-10 mRNA、p-STAT3 mRNA水平:与正常组比较,其余各组各时间点IL-10 mRNA、p-STAT3 mRNA水平均显著升高(P<0.01);与模型组比较,中药组各时间点IL-10 mRNA、p-STAT3 mRNA均显著升高(P<0.01)。⑤IL-10、p-STAT3蛋白表达:与正常组比较,其余各组各时间点IL-10、p-STAT3蛋白表达均显著升高(P<0.01);与模型组比较,中药组在各时间点IL-10、p-STAT3均升高(P<0.05)。结论:①慢加急性肝衰竭大鼠肝组织存在炎症因子表达失衡,以促炎因子占据主导地位。②温阳解毒化瘀方通过增加IL-10/STAT3表达促进抑炎因子分泌,使炎症反应由促炎向抑炎方向发展,这可能是其抗肝衰竭机制之一。

Objective:To investigate the regulation effect of the Wenyang Jiedu Huayu prescription(温阳解毒化瘀方)on the expression of IL-10/STAT3 and secretion of related inflammatory factors in liver tissue of chronic acute liver failure rats.Methods:Healthy male SD rats were randomly divided into the normal group,the model group and the TCM group.The acute-on-chronic liver failure(ACLF)rat model was established by multi-point subcutaneous injection of bovine serum albumin sensitization combined with D-Gal/LPS acute intraperitoneal attack.The normal group was injected intraperitoneally with equal dose of normal saline.The TCM group was given the Wenyang Jiedu Huayu prescription by intragastric administration,while the normal group and the model group were given equal dose of distilled water by intragastric administration.Liver function,histopathological changes of liver tissue,inflammatory factors(TNF-α,IL-1β,TGF-β,IL-10),STAT3 related protein and gene expression were detected at 1 h,12 h and 24 h after acute attack.Results:①Liver function:compared with the normal group,the contents of ALT and AST in other groups were significantly increased at each time point(P<0.01),compared with model group,ALT and AST contents in TCM group were significantly decreased at each time point(P<0.01).②Levels of inflammatory factors:compared with the normal group,the expressions of inflammatory factors(TNF-α,IL-1β,TGF-β,IL-10)in other groups were significantly increased at each time point(P<0.01),compared with the model group,the expressions of pro-inflammatory factors(TNF-α,IL-1β)in the TCM group were significantly decreased at each time point(P<0.01),while the expressions of anti-inflammatory factors(TGF-β,IL-10)were significantly increased(P<0.01).③Liver histopathological score:compared with the normal group,liver injury was significantly aggravated in other groups at each time point(P<0.01),compared with the model group,liver injury in the TCM group was reduced(P<0.05).④IL-10 mRNA and p-STAT3 mRNA levels:compared with the normal group,IL-10 mRNA and p-STAT3 mRNA levels in other groups at each time point were significantly increased(P<0.01),compared with the model group,IL-10 mRNA and P-STAT3 mRNA in the TCM group were significantly increased at each time point(P<0.01).⑤The expression of IL-10 and p-STAT3 protein:compared with the normal group,the expression of IL-10 and p-STAT3 protein at each time point in other groups were significantly increased(P<0.01),compared with the model group,IL-10 and p-STAT3 in the TCM group were increased at each time point(P<0.05).Conclusion:①The expression of inflammatory factors in ACLF rats is unbalanced,and pro-inflammatory factors play a dominant role.②The Wenyang Jiedu Huayu prescription promoted inflammatory response from pro-inflammatory to anti-inflammatory by increasing IL-10/STAT3 expression,which may be one of its anti-liver failure mechanisms.