基于Keap1-Nrf2信号通路研究补青颗粒对2型糖尿病大鼠肝损伤的保护作用及机制

Protective Effect and Mechanism of Buqing Granule on Liver Injury in Type 2 Diabetic Rats Based on Keap1-Nrf2 Signaling Pathway

目的基于Keap1-Nrf2信号通路探讨补青颗粒对2型糖尿病(type 2 diabetes mellitus,T2DM)大鼠肝损伤的保护作用及其机制。方法采用高脂高糖饲料联合小剂量链脲佐菌素制备T2DM模型后持续高脂高糖饲料喂养建立T2DM合并肝损伤大鼠模型。实验设置空白组、模型组、补青颗粒高剂量组、补青颗粒中剂量组,给药8周后取材检测各组大鼠血糖、肝功能、氧化应激相关指标;HE染色观察肝组织病理形态;Western blot、RT-qPCR法检测大鼠肝脏中Keap1-Nrf2通路相关蛋白表达水平及mRNA表达情况。结果与空白组比较,模型组大鼠体质量下降、血糖升高、肝功能受损伤(P均<0.05);补青颗粒组体质量、血糖及肝功能均较模型组改善(P均<0.05);补青颗粒组氧化应激相关指标均较模型组改善(P均<0.05);补青颗粒各剂量组Keap1、Cytoplasm Nrf2蛋白表达较模型组均减少,Nuclear Nrf2、NQO1、HO-1较模型组均升高(P均<0.05)。补青颗粒各组Keap1、Nrf2、NQO1、HO-1 mRNA表达与模型组比较均上调(P均<0.05)。结论补青颗粒可减轻T2DM大鼠肝损伤,改善大鼠一般状况及肝组织病理学改变,其机制可能与调控Keap1-Nrf2信号通路从而对抗氧化损伤有关。

Objective To investigate the protective effect of Buqing granule on liver injury in type 2 diabetes mellitus(T2DM)rats and its mechanism based on Keap1-Nrf2 signaling pathway.Methods T2DM was induced by high-fat and high-sugar diet combined with low-dose streptozotocin,and then the T2DM model with liver injury was established by continuous high-fat and high-sugar diet.The control group,model group,high-dose Buqing granule group,and middle-dose Buqing granule group were set up in the experiments.After 8 weeks of administration,the blood glucose,liver function,and oxidative stress related indexes of rats in each group were detected.HE staining was used to observe the pathological morphology of liver tissue.Western blot and RT-qPCR were used to detect the expression level of Keap1-Nrf2 pathway-related proteins and mRNA expression in rat liver.Results Compared with the control group,the rats in the model group had significant weight loss,blood sugar increase,and liver function damage(P all<0.05).The weight,blood sugar and liver function in the Buqing granule group were improved compared with the model group(P all<0.05).The oxidative stress-related indexes in the Buqing granule group were improved compared with the model group(P<0.05).The expression of Keap1 and Cytoplasm Nrf2 protein in each dose group of Buqing granule was lower than that in the model group,while the expressionlevels of Nuclear Nrf2,NQO1 and HO-1 were higher than those in the model group(P all<0.05).The expression of Nrf2,NQO1 and HO-1 mRNA in each group of Buqing granule was up-regulated compared with the model group(P all<0.05).Conclusion Buqing granule can reduce liver injury in T2DM rats,improve the general condition and pathological changes of liver tissue in rats,and its mechanism may be related to the regulation of Keap1-Nrf2 signaling pathway to combat antioxidant damage.