左归降糖清脂方含药血清对体外肝细胞损伤模型NLRP3信号通路的影响  

Effects of serum containing Zuogui Jiangtang Qingzhi Formula on NLRP3 signaling pathway in hepatocyte injury model in vitro

作　　者：曾广娴 喻嵘[2] 陈聪 向琴[2] 勾阳阳 ZENG Guang-xian;YU Rong;CHEN Cong;XIANG Qin;GOU Yang-yang(Qianxinan Hospital of Traditional Chinese Medicine,Xingyi 562400,China;Hunan University of Chinese Medicine,Changsha 410208,China;Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]黔西南州中医医院,贵州兴义562400 [2]湖南中医药大学,长沙410208 [3]贵州中医药大学,贵阳550025

出　　处：《中华中医药杂志》2023年第4期1823-1827,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81860818,No.82004185);贵州省科技厅科学技术基金项目(No.黔科合基础-ZK[2021]一般501);贵州省中医药管理局中医药、民族医药科学技术研究专项(No.QZYY-2022-010);2021贵州省卫生健康委科学技术基金项目;贵州中医药大学糖脂代谢病研究中心项目。

摘　　要：目的:探讨左归降糖清脂方含药血清对体外肝细胞损伤模型NLRP3信号通路的影响。方法:将BRL肝细胞随机分为正常组、模型组、空白血清组、中药血清组、阳性药物血清组,干预24 h后,油红O染色观察各组肝细胞内脂滴,CCK-8法检测肝细胞的增殖情况,常规生化法检测肝细胞上清液生化指标[谷丙转氨酶(ALT)、谷草转氨酶(AST)、甘油三酯(TG)、总胆固醇(TC)],ELISA测定肝细胞上清液NLRP3、Caspase-1、白细胞介素-1β(IL-1β)含量,分别采用qRT-PCR和Western Blot检测肝细胞NLRP3、ASC、Caspase-1 mRNA和蛋白表达。结果:与模型组比较,中药血清组脂滴蓄积明显减少;肝细胞上清液中ALT、AST、TG、TC含量均显著降低(P<0.01),NLRP3、Caspase-1、IL-1β含量均显著降低(P<0.01);肝细胞内NLRP3、ASC、Caspase-1 mRNA及蛋白表达水平显著下调(P<0.01)。结论:左归降糖清脂方含药血清可能通过抑制NLRP3炎症信号通路相关分子活化,改善肝细胞功能,从而发挥保护肝细胞的作用。Objective:To investigate the effect of serum containing Zuogui Jiangtang Qingzhi Formula on NLRP3 signaling pathway in hepatocyte injury model in vitro.Methods:BRL liver cells were randomly divided into normal group,model group,blank serum group,Zuogui Jiangtang Qingzhi Formula containing drug serum group and positive drug serum group.Twenty-four hours after intervention,lipid droplets in liver cells of each group were observed by oil red O staining,proliferation of liver cells was detected by CCK-8 method,biochemical indexes(ALT,AST,TG,TC)of supernatant of liver cells were detected by conventional biochemical method.The contents of NLRP3,Caspase-1 and IL-1βin supernatant of hepatocytes were determined by ELISA,and the mRNA or protein expressions of NLRP3,ASC and Caspase-1 in hepatocytes were detected by qRT-PCR or Western Blot.Results:Compared with the model group,the accumulation of lipid droplets in serum group containing Zuogui Jiangtang Qingzhi Formula was reduced,the concentration of biochemical indexes(ALT,AST,TG,TC)in supernatant of hepatocytes decreased(P<0.01),and the contents of NLRP3,Caspase-1 and IL-1βdecreased(P<0.01),the mRNA and protein expression levels of NLRP3,ASC and Caspase-1 in hepatocytes were significantly down-regulated(P<0.01).Conclusion:The serum containing Zuogui Jiangtang Qingzhi Formula may improve the function of liver cells by inhibiting the activation of NLRP3 inflammatory signaling pathway related molecules,and thus play a protective role in liver cells.

关 键 词：体外肝细胞损伤模型 左归降糖清脂方 含药血清 NLRP3信号通路 机制 2型糖尿病 非酒精性脂肪性肝病 

分 类 号：R285[医药卫生—中药学]