盐酸小檗碱通过调节氧化应激和炎症通路改善小鼠肝脏衰老

Berberine alleviates hepatic aging in mice by modulating oxidative stress and inflammatory

目的研究盐酸小檗碱对小鼠肝脏的抗衰老作用及可能的机制。方法取12只4周龄C57BL6/J小鼠,分为老年对照组(普通饲料喂养)、老年干预组(含有1 g/kg盐酸小檗碱饲料喂养),每组6只,饲养60周。小鼠64周龄时取材,肝组织石蜡切片进行HE染色观察小鼠肝脏组织病理变化;免疫荧光法检测肝脏组织中p16蛋白表达水平;比色法检测小鼠肝脏中丙二醛(malondialdehyde,MDA)的含量、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测肝脏组织中白细胞介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和血清中IL-8的表达;Western印迹法检测p16、p21、核因子E2相关因子2(nuclear factor erythroid-2 related factor 2,Nrf2)、核因子-κB(nuclear factor-κB,NF-κB)、磷酸化(phospho,p-)NF-κB、NF-κB抑制蛋白(inhibitoryκB,IκB)α、p-IκBα、p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)表达。同样饲养条件的普通饲料喂养16周龄小鼠作为年轻对照组。结果与年轻对照组相比,老年对照组肝脏组织形态老化,抗氧化物水能力降低(P<0.01),炎症因子水平明显上升(P<0.05或P<0.01);对比老年对照组,盐酸小檗碱干预可以明显改善肝脏衰老形态,衰老标记物p16和p21蛋白表达水平显著升高(P<0.05或P<0.01),提高小鼠抗氧化能力(P<0.05或P<0.01),降低炎症因子水平(P<0.05或P<0.01),下调p38 MAPK/NF-κB蛋白及相关因子Nrf2表达水平(P<0.001)。结论盐酸小檗碱改善老年小鼠肝组织的衰老形态,可能通过抑制p38 MAPK/NF-κB炎症信号通路降低炎症因子水平和抑制Nrf2通路改善衰老机体的氧化应激而发挥抗衰老作用。

Objective To explore the anti-aging effects of berberine hydrochloride and underlying mechanism.Methods Twelve 4-week-old C57BL6/J mice were divided into aging group(fed with normal chaw),aging+BBR intervention group(fed with normal chaw containing 1 g/kg berberine hydrochloride).At the age of 64 weeks,all the experimental mice were executed.The liver tissues were made into paraffin sections for HE staining to observe the morphological and structural integrity of liver parenchymal cells for pathological evaluation.Immunofluorescence was used to detect p16 protein expression levels in liver.The expression levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)were detected by colorimetry.The expression levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)in liver tissues and IL-8 in serum were detected by enzyme linked immunosorbent assay(ELISA)technique.The p16,p21,nuclear factor erythroid-2 related factor 2(Nrf2),nuclear factor-κB(NF-κB),phospho(p-)NF-κB,inhibitoryκB(IκB)α,p-IκBα,and p38 mitogen-activated protein kinase(MAPK)protein expression levels in liver were detected by Western blotting.The same indices were tested in the 16-week-old mice as the young control group.Results Compared with the young control group,the liver tissue in the aging control group exhibited morphological aging and antioxidant capacity were reduced(P<0.01),and inflammatory factors were increased(P<0.05 or P<0.01).Compared with the aging group,the liver tissues in the aging+BBR intervention group were still maintain regular arrangement of hepatocytes,the p16 and p21 protein expression levels were significantly increased(P<0.05 or P<0.01),the antioxidant capacity were increased(P<0.05 or P<0.01),the level of inflammatory factors were decresed(P<0.05 or P<0.01),and the p38 MAPK/NF-κB pathway and Nrf2 pathway were inhibited(P<0.001).Conclusion Berberine hydrochloride improves the aging morphology of the liver,potentially by suppressing the p38 MAPK/NF-κB pathway to reduce inflammation levels and inhibiting the Nrf2 pathway to ameliorate oxidative stress.