红花提取物通过调控Nrf2/STAT3/NF-κB信号通路对酒精性肝病的作用机制研究

Mechanism of Action of Carthamus tinctorius L.Extract on Alcoholic Liver Disease Through Modulation of Nrf2/STAT3/NF-κB Signaling Pathway

目的探讨红花提取物(Carthamus tinctorius L.extract,CTLE)对乙醇诱导的酒精性肝病小鼠氧化应激和炎症水平的影响及其作用机制。方法SPF级C57BL/6雄性小鼠随机分为4组:对照组、模型组、红花提取物低剂量组(50 mg·kg^(-1))、红花提取物高剂量组(100 mg·kg^(-1))。对照组给予Lieber-Decarli液体饲料,其他组给予Lieber-Decarli酒精饲料构建小鼠慢性酒精性肝损伤模型。收集小鼠血清和肝组织,检测小鼠血清生化指标。通过HE和油红O染色观察小鼠肝组织病理变化。qRT-PCR和Western Blot法检测Keap1/Nrf2和STAT3/NF-κB通路相关因子的mRNA和蛋白表达水平。结果与模型组比较,给药组丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、低密度脂蛋白胆固醇(LDL-C)、丙二醛(MDA)的水平明显降低(P<0.05,P<0.01),而高密度脂蛋白胆固醇(HDL-C)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)的水平明显升高(P<0.05,P<0.01),表明红花提取物对小鼠酒精性肝损伤具有一定的保护和抗氧化作用;HE染色和油红O染色观察到小鼠肝脏病变和脂质沉积有所改善。并且通过激活Keap1/Nrf2通路相关抗氧化因子的mRNA和蛋白表达水平,抑制STAT3/NF-κB通路及相关炎症因子的mRNA和蛋白表达水平(P<0.05,P<0.01),从而增强机体的抗氧化和抗炎作用。结论红花提取物可以通过调节Keap1/Nrf2和STAT3/NF-κB信号通路发挥抗氧化应激和抗炎作用,减轻小鼠的酒精性肝损伤,为治疗酒精性肝病和后续分子机制研究提供新的思路。

Objective To investigate the effects of Carthamus tinctorius L.extract(CTLE)on the levels of oxidative stress and inflammation in mice with ethanol-induced alcoholic liver disease and its mechanism of action.Methods SPF-grade C57BL/6 male mice were randomly divided into four groups:control group,model group,low-CTLE group(50 mg·kg^(-1)),and high-CTLE group(100 mg·kg^(-1)).The control group was given Lieber-Decarli liquid diet,and the other groups were given Lieber-Decarli alcohol diet to construct a chronic alcoholic liver injury model in mice.Serum and liver tissues of mice were collected and serum biochemical indexes of mice were detected.HE and oil red O staining were applied to observe pathological changes in mouse liver tissues.Real-time fluorescence quantitative PCR and Western Blot were used to detect the mRNA and protein expression levels of Keap1/Nrf2 and STAT3/NF-κB pathway-related factors.Results Compared with the model group,the ALT,AST,LDL-C,and MDA levels were significantly reduced(P<0.05,P<0.01),while the levels of HDL-C,SOD,and GSH were increased dramatically in the administered group(P<0.05,P<0.01),which indicated that CTLE has specific protective and antioxidant effects on alcoholic liver injury in mice.HE staining and oil red O staining showed that the hepatic lesions and lipid deposition of mice were ameliorated.It enhances the antioxidant and anti-inflammatory effects of the body by activating the mRNA and protein expression levels of antioxidant factors related to the Keap1/Nrf2 pathway and inhibiting the mRNA and protein expression levels of inflammatory factors related to STAT3/NFκB pathway(P<0.05,P<0.01).Conclusion It was shown that CTLE could exert anti-oxidative stress and anti-inflammatory effects through regulating Keap1/Nrf2 and STAT3/NF-κB signaling pathways to attenuate alcoholic liver injury in mice.This study may provide a new idea for the treatment of alcoholic liver disease and the subsequent study of molecular mechanisms.