固本延龄丸通过NF-κB信号通路拮抗小鼠肝衰老作用及其机制研究

Effect of Guben Yanling pills in antagonising liver aging in mice through NF-κB signaling pathway and its mechanism

目的探讨固本延龄丸对衰老小鼠肝衰老的影响及其作用机制。方法小鼠随机分为空白对照组、模型组、维生素E组(0.1 g·kg^(-1))及固本延龄丸低、中、高剂量组(0.59、1.17、2.34 g·kg^(-1)),颈背部皮下注射D-半乳糖(150 mg·kg^(-1))建立衰老小鼠模型,造模的同时,各组灌胃给予相应溶媒或药物,每日1次,连续6周。检测各组小鼠主要脏器指数;HE染色观察肝组织形态学;比色法检测肝中SA-β-Gal活力;ELISA法检测血清TNF-α、IL-1β、IL-6、IL-4和IL-10含量;Western blot法检测肝中IKKβ、IκBα和NF-κB p65蛋白相对表达水平;免疫荧光法检测肝中NF-κB p65入核表达水平。结果与空白对照组比较,模型组小鼠的脑、肝、肾、脾及胸腺的脏器指数下降(P<0.05,P<0.01);肝中SA-β-Gal活力升高(P<0.01);肝组织形态结构受损明显;TNF-α、IL-1β和IL-6含量升高(P<0.01)、IL-4和IL-10含量降低(P<0.01);IKKβ、NF-κB p65表达水平升高(P<0.01)、IκBα表达水平降低(P<0.01);细胞核内NF-κB p65表达水平升高(P<0.01)。与模型组比较,固本延龄丸各剂量组小鼠的脑、肝、肾、脾及胸腺的脏器指数升高(P<0.05,P<0.01);肝中SA-β-Gal活力降低(P<0.01);肝组织形态结构受损明显改善;TNF-α、IL-1β和IL-6含量降低(P<0.01);IL-4和IL-10含量升高(P<0.01);IKKβ、NF-κB p65表达水平降低(P<0.01)、IκBα表达水平升高(P<0.01);细胞核内NF-κB p65表达水平降低(P<0.01)。结论固本延龄丸能够拮抗小鼠的肝衰老,其作用机制可能与抑制肝中NF-κB信号通路的激活、下调其下游促炎因子水平并上调抑炎因子水平、减轻肝内炎症反应有关。

Aim To study the effect of Guben Yanling pills on liver aging in aging mice and the related mechanism.Methods The mice were randomly divided into blank control group,model group,vitamin E group(0.1 g·kg^(-1))and low,medium and high dose groups(0.59,1.17,2.34 g·kg^(-1))of Guben Yanling pills.The aging mouse model was established by subcutaneous injection of D-galactose(150 mg·kg^(-1))into the back of neck.At the same time of modeling,the corresponding drugs were given by gavage once a day for six weeks.The main organ indexes were calculated.HE staining was used to observe the morphology of liver tissue.Colorimetry was used to detect the activity ofβ-galactosidase in liver.ELISA was used to detect the content of TNF-α,IL-1β,IL-6,IL-4,IL-10.Western blot was used to detect the protein relative expression level of IKKβ,IκBα,NF-κB p65.Immunofluorescence was used to detect the expression level of NF-κB p65.Results Compared with the blank control group,the organ index of the brain,liver,kidney,spleen,and thymus in the model group decreased(P<0.05,P<0.01),the activity ofβ-galactosidase increased(P<0.01),liver tissue morphology and structure were significantly damaged,the content of TNF-α,IL-1βand IL-6 increased(P<0.01),the content of IL-4 and IL-10 decreased(P<0.01),the levels of IKKβ,NF-κB p65 increased(P<0.01),the levels of IκBαdecreased(P<0.01),and the levels of NF-κB p65 in nucleus increased(P<0.01).Compared with the model group,the organ indexes of brain,liver,kidney,spleen,and thymus in each dose group of Guben Yanling pills increased(P<0.05,P<0.01),the activity ofβ-galactosidase decreased(P<0.01),the morphological and structural damage of liver tissue was significantly improved,the content of TNF-α,IL-1βand IL-6 decreased(P<0.01),the content of IL-4 and IL-10 increased(P<0.01),the levels of IKKβ,NF-κB p65 decreased(P<0.01),the levels of IκBαincreased(P<0.01),and the levels of NF-κB p65 in nucleus decreased(P<0.01).Conclusions Guben Yanling pills can antagonize liver aging in mice,and its mechanism may be related to inhibiting the activation of NF-κB signaling pathway in liver,downregulating downstream pro-inflammatory factor levels,upregulating anti-inflammatory factor levels,and alleviating inflammation in liver.