Objective: To observe the therapeutic effect of Shengsan Jiedu Huayu decoction in alleviating inflammatory liver injury in rats with acute-on-chronic liver failure (ACLF) and its effect on the activation intensity for the NLRP3 signaling pathway. Methods: 63 SD rats were randomly divided into a blank group, a model group, and low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction (7.29 g/kg/d, 14.58 g/kg/d, and 29.16 g/kg/d). The ACLF rat model was replicated using carbon tetrachloride combined with d-galactosamine and lipopolysaccharide. Different dose gradients of the Shengsan Jiedu Huayu decoction were used for a five-day intervention treatment, and then rat serum and tissue samples were collected. A biochemical analyzer was used to detect the serum levels of ALT, AST, and TBIL in rats. ELISA was used to detect serum IL-18 and IL-1β content. HE staining was used to observe histomorphological changes in liver tissue. Immunohistochemistry was used to detect GSDMD expression in liver tissue. Western blot and PCR were used to detect NLRP3, Caspase1, ASC, TLR4, IL-1β, IL-18 protein, and mRNA expression levels.The groups were compared using analysis of variance and the rank-sum test. Results: Compared with the blank group, the model group's rat liver tissue was severely injured. Serum levels of ALT, AST, and TBIL, inflammatory factors IL-1β and IL-18, and the GSDMD protein expression level, NLRP3 expression level, TLR4, caspase 1, ASC, IL-1β, IL-18 protein, and mRNA (P<0.01) were all significantly increased in the model than the blank group (P<0.01). Additionally, compared with the model group, the low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction had improved liver tissue injury in ACLF rats, while the serum levels of ALT, AST, TBIL, IL-1β, IL-18, liver tissue GSDMD protein, NLRP3, TLR4, caspase 1, and ASC expressions were all lower in the different dose gradients of the Shengsan Jiedu Huayu decoction than the model group, with the most evident reduction in the high-dose group (P<0.01). Conclusion: Shengsan Jiedu Huayu decoction can weaken the activation intensity of the NLRP3 signaling pathway, alleviate liver tissue pathological injury, reduce inflammatory factor release, and alleviate inflammatory liver injury in ACLF rats.
目的： 观察解毒化瘀升散方缓解慢加急性肝衰竭（ACLF）大鼠肝脏炎症损伤的疗效及其对NLRP3信号通路活化强度的影响。 方法： 取SD大鼠63只，随机分为空白组、模型组和解毒化瘀升散方低、中、高剂量组（分别为7.29、14.58及29.16 g·kg(-1)·d(-1)）。采用四氯化碳联合D-氨基半乳糖和脂多糖复制ACLF大鼠模型，解毒化瘀升散方不同剂量梯度干预处理5 d，收集大鼠血清及组织样本。采用生物化学分析仪检测大鼠血清丙氨酸转氨酶、天冬氨酸转氨酶、总胆红素水平；酶联免疫吸附法检测血清白细胞介素-18、白细胞介素-1β含量；苏木精-伊红染色观察肝组织形态学改变；免疫组织化学法检测肝组织中消皮素D (GSDMD)表达；蛋白质免疫印迹法及PCR检测NLRP3、Caspase-1、凋亡相关斑点样蛋白、Toll样受体4、白细胞介素-1β、白细胞介素-18mRNA和蛋白表达水平。计量资料采用方差分析，组间比较采用秩和检验。 结果： 与空白组相比，模型组大鼠肝组织损伤严重，血清丙氨酸转氨酶、天冬氨酸转氨酶、总胆红素水平，炎症因子白细胞介素-1β、白细胞介素-18水平，GSDMD蛋白表达水平，均较空白组显著升高（P值均<0.01），NLRP3、TLR4、Caspase 1、凋亡相关斑点样蛋白、白细胞介素-1β、白细胞介素-18蛋白与mRNA表达水平也显著升高（P值均 < 0.01）；与模型组相比，解毒化瘀升散方低、中、高剂量组能改善ACLF大鼠肝组织损伤，血清丙氨酸转氨酶、天冬氨酸转氨酶、总胆红素水平，血清白细胞介素-1β、白细胞介素-18水平，肝组织GSDMD蛋白水平，NLRP3、TLR4、Caspase 1、凋亡相关斑点样蛋白表达均较模型组降低，其中以高剂量组降低程度最明显（P 值均< 0.01）。 结论： 解毒化瘀升散方能减弱ACLF大鼠肝脏NLRP3信号通路活化强度，减轻肝组织病理损伤，减少炎症因子释放，缓解ACLF大鼠肝脏炎症损伤。.
Keywords: Acute-on-chronic liver failure; Inflammation; Jiedu Huayu Shengsan Formula; NLRP3 signaling pathway.