山楂原花青素和VC联合通过Wnt/β-catenin通路减轻胰岛素抵抗大鼠肝脏氧化应激  

Hawthorn Proanthocyanidins and Vitamin C Attenuate Liver Oxidative Stress in Insulin Resistance Rats through the Wnt/β-catenin Signaling Pathway

作　　者：李西栋[1] 孙绍霞[1] 梁亚楠 石塔拉 宓伟 LI Xidong;SUN Shaoxia;LIANG Yanan;SHI Tala;MI Wei(Cardiovascular Internal Medicine Department,Linyi People’s Hospital,Linyi 276000,China;School of Public Health and Management,Binzhou Medical University,Yantai 264003,China)

机构地区：[1]临沂市人民医院心血管内科,山东临沂276000 [2]滨州医学院公共卫生与管理学院,山东烟台264003

出　　处：《食品科学》2021年第17期186-192,共7页Food Science

基　　金：山东省自然科学基金项目(2017LH059);山东省高等学校科技计划项目(J17KA224)。

摘　　要：目的:探讨山楂原花青素(hawthorn proanthocyanidin,HPC)和VC联合通过Wnt/β-catenin通路减轻胰岛素抵抗大鼠氧化应激的机制。方法:80只雄性Wistar大鼠通过高脂膳食法建立胰岛素抵抗模型,10只饲喂普通饲料作为对照组,检测大鼠造模前后体质量变化和造模后空腹血糖、血清胰岛素浓度,比色法检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)和碱性磷酸酶(alkaline phosphatase,ALP)活力;随机选取50只造模成功的胰岛素抵抗大鼠分为模型组、HPC组(56μg/mL)、VC组(180μg/mL)、HPC(56μg/mL)+VC(180μg/mL)组和二甲双胍(2μg/mL)组,每组各10只。连续给药12周,测定各组大鼠血液中葡萄糖、胰岛素浓度以及肝匀浆中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)、还原型谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)的水平;实时定量聚合酶链式反应检测Wnt1、轴蛋白(Axin)、糖原合酶激酶-3β(glycogen synthasc kinase-3β,GSK-3β)、腺瘤样息肉病蛋白(adenomatous polyposis coli protein,APC)、β-catenin、过氧化物酶体增殖物激活受体γ(peroxisome proliferators-activated receptorsγ,PPARγ)的mRNA表达水平;蛋白质印迹法检测肝组织Wnt1、β-catenin和PPARγ的蛋白表达水平;荧光共振能量转移分析法检测Wnt1和Dsh蛋白结合的情况。结果:80只大鼠给予高脂膳食喂养,有54只造模成功,成功率67.5%。造模后,模型组大鼠的体质量以及空腹血糖、血清胰岛素、ALT、AST和ALP水平均极显著高于对照组大鼠(P<0.01)。与模型组相比,HPC组、HPC+VC组和二甲双胍组大鼠肝匀浆中各指标水平均有极显著差异(P<0.01),HPC+VC组对肝匀浆中各指标干预效果显著优于HPC组(P<0.05);HPC+VC组Wnt1和β-catenin的mRNA相对表达水平极显著降低(P<0.01),Axin、GSK-3β、APC和PPARγ的mRNA相对表To explore the mechanism by which hawthorn proanthocyanidins(HPC)combined with vitamin C(VC)alleviates liver oxidative stress in insulin resistance(IR)rats through the Wnt/β-catenin signaling pathway.To create an animal model of IR,80 Wistar male rats were given a high-fat diet.The changes in body mass,fasting blood glucose and serum insulin concentration were measured before and after the establishment of the model.Colorimetry was used to test the activities of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP).A total of 50 IR rats were randomly selected and divided into model,HPC(56μg/mL),VC(180μg/mL),HPC(56μg/mL)+VC(180μg/mL)and metformin(2μg/mL)groups,each consisting of 10 animals.Blood glucose and insulin levels,as well as the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GPx),glutathione(GSH)and the content of malondialdehyde(MDA)in liver homogenate were determined after 12 weeks of continuous administration.The mRNA expression of Wnt1,Axin,glycogen synthasc kinase-3β(GSK-3β),adenomatous polyposis coli protein(APC),β-catenin and peroxisome proliferators-activated receptorsγ(PPARγ)were tested by real-time polymerase chain reaction(PCR).The expression of Wnt1,β-catenin and PPARγproteins was tested by Western blotting.The binding capacity of Wnt1 to dishevelled(Dsh)proteins was evaluated by fluorescence resonance energy transfer(FRET)assay.IR was successfully induced in 54(67.5%)of the 80 rats.The body mass,fasting blood glucose,serum insulin,and ALT,AST and ALP activities of the model group were significantly higher than those of the control group(P<0.01).Compared with the model group,the three intervention groups exhibited significant differences in all the studied parameters in the liver homogenate(P<0.01),and the effect of HPC+VC was better than that of HPC alone(P<0.05).In the HPC+VC group,the relative mRNA expression of Wnt1 andβ-catenin was decreased significantly(P<0.01),while the relative mRNA expression of Axi

关 键 词：山楂原花青素 VC WNT/Β-CATENIN通路 胰岛素抵抗 肝脏氧化应激 

分 类 号：R151.1[医药卫生—营养与食品卫生学]