基于NLRP3/ASC/Caspase-1信号通路探讨肝复胶囊抗慢性肝损伤的作用机制  

Effect and mechanism of Ganfu capsule against chronic liver injury based on NLRP3/ASC/Caspase⁃1 signaling pathway

作　　者：李敏仪 黄以蓉 彭凯 彭继 谢锋 王祁 高璐 李敏[1] LI Minyi;HUANG Yirong;PENG Kai;PENG Ji;XIE Feng;WANG Qi;GAO Lu;LI Min(Clinical Chinese Pharmacy Teaching and Research Section,School of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Pharmacy,Ankang Hospital of Traditional Chinese Medicine)

机构地区：[1]陕西中医药大学药学院临床中药学教研室,咸阳712046 [2]安康市中医医院药学部

出　　处：《山西医科大学学报》2025年第1期28-35,共8页Journal of Shanxi Medical University

基　　金：秦创原中药创新研发项目(2022-QCYZH-050);陕西中医药大学校级高水平中医药重点学科建设-临床中药学项目(2024XKZD12)。

摘　　要：目的 研究肝复胶囊(Ganfu capsule,GFC)对CCl4诱导的慢性肝损伤(chronic liver injury,CLI)大鼠模型的影响及作用机制。方法 将60只SD雄性大鼠随机分为空白组、模型组、秋水仙碱组(2×10^(-4) g/kg)及肝复胶囊低(GFC-L,0.781 3 g/kg)、中(GFC-M,1.562 5 g/kg)、高(GFC-H,3.125 g/kg)剂量组,每组10只。空白组不做处理,其余组大鼠均背部皮下注射40%CCl4花生油混合溶液造模,除首剂量按5 mL/kg外,每次注射剂量为3 mL/kg,每周2次,持续12周。造模4周后,给药组大鼠开始灌胃相应药物,空白组和模型组灌胃等量生理盐水,每日1次,持续8周。12周末,称量各组大鼠体质量后,处死大鼠,腹主动脉取血,收集大鼠肝脏。HE及Masson染色观察肝组织病理变化;生化试剂盒检测大鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST);ELISA检测大鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、IL-6水平;Western blot法检测各组大鼠肝组织中核苷酸结合寡聚化结构域样受体蛋白3(nod-like receptor protein 3,NLRP3)、凋亡相关斑点样蛋白(apoptosis associated speck-like protein containing a caspase activating recruitment domain,ASC)、半胱氨酸蛋白酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)蛋白表达。结果 与模型组比较,GFC各剂量组和秋水仙碱组大鼠体质量显著增加,肝组织炎症浸润及胶原蛋白沉积减少,血清中ALT、AST、TNF-α、IL-1β、IL-6水平明显降低(P<0.05),肝组织中NLRP3、ASC、Caspase-1蛋白表达下调(P<0.05),高剂量GFC改善CLI大鼠疗效优于秋水仙碱。结论 GFC可能通过下调NLRP3/ASC/Caspase-1信号通路,抑制炎症因子的表达,从而缓解CCl4诱导的肝脏炎症损伤,发挥保护肝脏作用。Objective To study the effect and mechanism of Ganfu capsule(GFC)against CCl4-induced chronic liver injury(CLI)in rats.Methods Sixty SD male rats were randomly divided into control group,model group,Colchicine group(2×10^(-4) g/kg)and Ganfu capsule low dose group(GFC-L,0.7813 g/kg),medium dose group(GFC-M,1.5625 g/kg),and high dose group(GFC-H,3.125 g/kg),with 10 rats in each group.The rats in control group were not treated,and the rats in the other groups were subcuta-neously injected with 40%CCl4 peanut oil mixture(3 mL/kg,except for the first dose of 5 mL/kg)on the dorsal surface twice a week for 12 weeks.At week 4,the rats in drug groups were gavaged with the corresponding drugs once a day,and the rats in control group and model group were gavaged with the same amount of saline.At the end of 12 weeks,the rats were weighed and then sacrificed to collect the blood from the abdominal aorta and the livers.HE staining and Masson staining were used to observe the pathological changes of liver tissues.Biochemical kits were used to detect serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST).ELISA was used to detect the expressions of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6.Western blot was used to detect the protein expression levels of nod-like receptor protein 3(NLRP3),apoptosis associated speck-like protein contain-ing a caspase activating recruitment domain(ASC),and cysteinyl aspartate specific proteinase-1(Caspase-1).Results Compared with model group,the body mass was significantly increased in GFC groups and Colchicine group,the inflammatory infiltration and collagen deposition in liver tissues were reduced,the serum levels of ALT,AST,TNF-α,IL-1β,and IL-6 were decreased(P<0.05),and the protein expressions of NLRP3,ASC,and Caspase-1 in liver tissues were downregulated(P<0.05),and the efficacy of high dose of GFC for improving CLI was better than that of Colchicine.Conclusion GFC may alleviate the CCl4-induced hepatic inflam-matory injury and exert hepatopr

关 键 词：肝复胶囊 慢性肝损伤 NLRP3 ASC CASPASE-1 炎症 

分 类 号：R575[医药卫生—消化系统]