褐藻胶寡糖对D-半乳糖诱导的衰老模型小鼠肝脏损伤的保护作用及其机制  

PROTECTIVE EFFECT OF ALGINATE OLIGOSACCHARIDES AGAINST D-GALACTOSE-INDUCED LIVER INJURY IN A MOUSE MODEL OF SENESCENCE AND ITS MECHANISM

作　　者：陈茗 冯文静[1] 胡松[1] 刘佳[1] 王珊 毛拥军[1] CHEN Ming;FENG Wenjing;HU Song;LIU Jia;WANG Shan;MAO Yongjun(Department of Geriatric Medicine,The Affiliated Hospital of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属医院老年医学科,山东青岛266071

出　　处：《精准医学杂志》2022年第3期217-221,共5页Journal of Precision Medicine

基　　金：山东省自然科学基金面上项目(ZR2021MH-197);山东省自然科学基金面上项目(ZR201911060375);中国博士后科学基金会面上项目一等资助(2018M630757)。

摘　　要：目的探讨褐藻胶寡糖(AOS)对D-半乳糖(D-gal)诱导的衰老模型小鼠肝脏损伤的保护作用及其可能的机制。方法将32只雄性C57BL/6J小鼠随机分为对照组(A组)、AOS组(B组)、D-gal组(C组)和D-gal+AOS组(D组)。C、D组小鼠颈背部皮下注射D-gal[200 mg/(kg·d)],其余各组注射等量生理盐水,连续8周。从第5周开始B、D组小鼠用AOS[200 mg/(kg·d)]进行灌胃处理,其余各组用等量生理盐水灌胃处理,连续4周。实验结束后,收集小鼠血液和肝脏组织样本,测定小鼠血清中ALT、AST的水平,检测肝脏组织中衰老相关蛋白p21和还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)的氧化酶亚基gp91 phox、p67 phox的表达情况,以及核因子E2相关因子2(Nrf 2)、血红素氧化酶-1(HO-1)mRNA和超氧化物歧化酶1(SOD1)蛋白的含量。结果小鼠血清中ALT、AST和肝脏组织中衰老蛋白p21检测结果显示,单用D-gal诱导小鼠肝脏衰老和肝脏损伤的效果最显著(F=77.05~836.38,P<0.05),AOS对D-gal诱导的小鼠肝脏衰老和肝脏损伤有改善作用(F=20.29~138.67,P<0.05)。Western blot检测结果显示,单用D-gal可提高小鼠肝脏组织中NADPH的氧化酶亚基gp91 phox、p67 phox的水平(F=690.11、66.68,P<0.05),降低抗氧化酶SOD1的水平(F=24.85,P<0.05),而AOS可抑制D-gal的作用(F=12.28~123.18,P<0.05)。RT-qPCR检测结果显示,单用D-gal可明显抑制肝脏组织中Nrf 2、HO-1 mRNA的表达(F=380.07、253.34,P<0.05),而AOS可抑制D-gal的作用(F=62.04、41.21,P<0.05)。结论AOS可有效抑制D-gal诱导的衰老小鼠肝脏损伤,保护肝脏的生理功能,其作用机制可能与调控肝脏细胞中Nrf2/HO-1信号通路,抑制肝脏组织中的氧化应激有关。Objective To investigate the protective effect of alginate oligosaccharides(AOS)against D-galactose(D-gal)-induced liver injury in a mouse model of senescence and its possible mechanism.Methods A total of 32 male C57BL/6J mice were randomly divided into control group(group A),AOS group(group B),D-gal group(group C),and D-gal+AOS group(group D).The mice in groups C and D were given subcutaneously injected D-gal[200 mg/(kg·d)]at the neck and back,and those in the other groups were given injection of an equal volume of normal saline,for 8 consecutive weeks.Since week 5,the mice in groups B and D were given AOS[200 mg/(kg·d)]by gavage,and those in the other groups were given an equal volume of normal saline by gavage,for 4 consecutive weeks.After the experiment ended,blood and liver tissue samples were collected to mea-sure the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)and the expression of the senescence-related proteins p21 and oxidase subunits gp91 phox and p67 phox of reduced nicotinamide adenine dinucleotide phosphate(NADPH)in liver tissue,as well as the mRNA expression levels of nuclear factor erythroid 2-related factor 2(Nrf 2)and heme oxygenase-1(HO-1)and the protein expression level of superoxide dismutase-1(SOD1).Results Measurement of the serum levels of ALT and AST and the expression of the senescence-related protein p21 in liver tissue showed that D-gal alone had the most significant effect in inducing liver senescence and liver injury in mice(F=77.05-836.38,P<0.05),and AOS improved liver senescence and liver injury induced by D-gal in mice(F=20.29-138.67,P<0.05).Western blot showed that D-gal alone increased the expression levels of NADPH oxidase subunits gp91 phox and p67 phox(F=690.11,66.68,P<0.05)and reduced the expression level of the antioxidant enzyme SOD1(F=24.85,P<0.05)in liver tissue,while AOS inhibited such effect of D-gal(F=12.28-123.18,P<0.05).RT-qPCR showed that D-gal alone significantly inhibited the mRNA expression levels of Nrf 2 and HO-1 in liver

关 键 词：褐藻酸 寡糖类 半乳糖 化学性与药物性肝损伤 衰老 氧化性应激 疾病模型 动物 

分 类 号：R575[医药卫生—消化系统] R916.4[医药卫生—内科学]