羟基红花黄色素A对非酒精性脂肪肝炎细胞的作用

The Effect of Hydroxysafflor Yellow A on Non-Alcoholic Steatohepatitis Cells

目的:探讨羟基红花黄色素A对非酒精性脂肪肝炎(NASH)的影响及其作用机制。方法:实验分为对照组、模型组和羟基红花黄色素A组,模型组用500μmol/L的游离脂肪酸(FFA)处理细胞24 h,对照组为不做处理的人正常肝脏细胞L-02,羟基红花黄色素A组在模型组基础上用0.2 mg/mL的羟基红花黄色素A处理细胞24 h。通过油红O染色法观察各组细胞内脂质沉积情况。ELISA检测各组细胞内谷丙转氨酶(ALT)、谷草转氨酶(AST)、超氧化物歧化酶(SOD)、丙二醛(MDA)、白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)含量。RT-qPCR检测细胞中炎症小体感受器分子NOD样受体蛋白3(NLRP3)mRNA、半胱氨酸蛋白酶-1(Caspase-1)mRNA、消皮素D(GSDMD)mRNA表达水平。Western blotting检测各组细胞中NLRP3、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达情况。结果:与对照组比较,模型组细胞变圆,细胞间隙变大,红色脂滴较多,ALT、AST、MDA、IL-18、IL-1β、NLRP3、Caspase-1、GSDMD含量明显增加,SOD含量明显下降(P<0.05);与模型组比较,羟基红花黄色素A组细胞红色脂滴减少,细胞内ALT、AST、MDA、IL-18、IL-1β、NLRP3、Caspase-1、GSDMD含量明显下降,SOD含量明显增加(P<0.05)。结论:羟基红花黄色素A能够减轻炎症反应和氧化应激反应,抑制细胞焦亡,从而改善非酒精性脂肪肝炎。

Objective:To investigate the effect of hydroxysafflor yellow A on non-alcoholic steatohepatitis(NASH)and its mechanism.Methods:The experiment was divided into control group,model group and hydroxysafflor yellow A group.The model group was treated with 500μmol/L free fatty acid(FFA)for 24 h,the control group was untreated human normal liver cells L-02,and the hydroxysafflor yellow A group was treated wit h0.2 mg/mL hydroxysafflor yellow A for 24 h on the basis of the model group.The oil red O staining method was used to observe the lipid deposition in the cells of each group.The levels of intracellular alanine aminotransferase(ALT),aspartate aminotransferase(AST),superoxide dismutase(SOD),malondialdehyde(MDA),interleukin-18(IL-18),interleukin-1β(IL-1β)were detected by ELISA.RT-qPCR was used to detect the expression of inflammasome receptor molecule NOD-like receptor protein 3(NLRP3)mRNA,cysteine protease-1(Caspase-1)mRNA,and steroid D(GSDMD)mRNA.Western blotting detected the protein expression of NLRP3,Caspase-1,GSDMD,IL-1β,and IL-18 in each group of cells.Results:Compared with the control group,the cells in the model group became rounded,the intercellular spaces became larger and there were more red lipid droplets.The contents of ALT,AST,MDA,IL-18,IL-1β,NLRP3,Caspase-1 and GSDMD were significantly increased,and The content of SOD was significantly decreased(P<0.05).Compared with the model group,the red lipid droplets in the hydroxysafflor yellow A group were reduced,the content of intracellular ALT,AST,MDA,IL-18,IL-1β,NLRP3,Caspase-1,and GSDMD were significantly decreased,and the content of SOD was significantly increased(P<0.05).Conclusion:Hydroxysafflor yellow A can reduce inflammation and oxidative stress,inhibit pyrolysis,and improve non-alcoholic steatohepatitis.