茯苓多糖提取物调控CYP2E1及NF-κB炎症通路改善小鼠酒精性肝病

建立酒精性肝病(alcoholic liver disease,ALD)模型,研究茯苓多糖(Poria cocos polysaccharides,PCP)提取物对ALD小鼠细胞色素P4502E1(CYP2E1)及核因子κB(NF-κB)炎症信号通路的影响,探讨其对ALD的保护作用及机制。60只SPF级雄性C57BL/6N小鼠随机分为对照组、模型组、阳性药组(联苯双酯200 mg·kg^-1)、茯苓多糖低剂量组(50 mg·kg^-1)、茯苓多糖高剂量组(200 mg·kg^-1),建立Gao-binge模型并给药。观察肝脏形态及病理学变化,计算脏器指数,检测各组小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,试剂盒分别测定小鼠肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)含量,ELISA法检测白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的水平,免疫荧光染色观察巨噬细胞激活情况,Western blot分析主要代谢酶CYP2E1及Toll样受体4(TLR4)、磷酸化核因子κB p65(p-NF-κB p65)、核因子κB p65(NF-κB p65)蛋白的表达。与模型组相比,PCP给药组可显著改善肝组织病理损伤情况。免疫荧光结果显示,与模型组相比,给药组肝组织炎性巨噬细胞数量减少,并且各给药剂量组ALT、AST、MDA、IL-6、TNF-α的含量显著降低(P<0.05),SOD活性显著升高(P<0.01)。茯苓多糖提取物对小鼠酒精性肝损伤具有保护作用,其机制可能是调控代谢酶CYP2E1的表达和抑制TLR4/NF-κB炎症信号通路,减轻氧化应激和炎症损伤,抑制ALD的发展。

Extracts of Poria cocos polysaccharides improves alcoholic liver disease in mice via CYP2E1 and NF-κB inflammatory pathways

The present study investigated the effect of extract of Poria cocos polysaccharides(PCP)on cytochrome P4502 E1(CYP2 E1)and nuclear factorκB(NF-κB)inflammatory signaling pathways in alcoholic liver disease(ALD)mice and explored its protective effect and mechanism.Sixty male C57 BL/6 N mice of SPF grade were randomly divided into a control group,a model group,a positive drug group(bifendate,200 mg·kg^-1),and high-(200 mg·kg^-1)and low-dose(50 mg·kg^-1)PCP groups.Gao-binge mo-del was induced and the mice in each group were treated correspondingly.Liver morphological and pathological changes were observed and organ index was calculated.Serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected.Malondialdehyde(MDA)and superoxide dismutase(SOD)in liver tissues were detected by assay kits.The levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by ELISA.The activation of macrophages was observed by immunofluorescence staining and protein expression of CYP2 E1,Toll-like receptor 4(TLR4),NF-κB p65,and phosphorylated NF-κB p65(p-NF-κB p65)were analyzed by Western blot.The ALD model was properly induced.Compared with the model group,the PCP groups significantly improved the pathological injury of liver tissues.Immunofluorescence staining revealed that compared with the model group,the groups with drug intervention showed decreased macrophages in liver tissues.Additionally,the PCP groups showed reduced ALT,AST,MDA,IL-6,and TNF-α(P<0.05),and potentiated activity of SOD(P<0.01).PCP extract has the protective effect against alcoholic liver injury in mice,and the underlying mechanism may be related to the regulation of the expression of CYP2 E1 and inhibition of TLR4/NF-κB inflammatory signaling pathway to reduce oxidative stress and inflammatory injury,thereby inhibiting the development of ALD.