CNPY2调控内质网应激PERK/eIF2α通路在有氧运动干预非酒精性脂肪性肝病中的作用  

Aerobic Exercise Regulates PERK/eIF2α Pathway via CNPY2 in Models of Non-alcoholic Fatty Liver Disease

作　　者：李军汉[1] 王佳倩 李亚龙 蒋昌君 Li Junhan;Wang Jiaxian;Li Yalong;Jiang Changjun(School of Sports Medicine and Health,Chengdu Sport University,Chengdu 610041,China)

机构地区：[1]成都体育学院运动医学与健康学院,成都610041

出　　处：《中国运动医学杂志》2022年第10期783-792,共10页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金青年基金资助项目(31900846);成都体育学院运动医学四川省重点实验室资助项目(CX21B04)。

摘　　要：目的:探讨有氧运动通过冠层成纤维细胞生长因子信号调节器2(CNPY2)调控内质网应激在PKR样内质网状激酶(PERK)/真核细胞翻译起始因子2α(eIF2α)通路改善非酒精性脂肪性肝病(NAFLD)中的可能机制。方法:30只8周龄雄性C57BL/6J小鼠随机分为普通饮食组(普饮组)、高脂饮食组(高脂组)和高脂饮食运动组(高脂运动组),每组10只。普饮组给予普通饲料喂养,高脂组和高脂运动组给予高脂饲料喂养,连续喂养18周,直至实验结束,取小鼠肝脏。从第10周开始,高脂运动组小鼠进行有氧跑台训练干预(12 m/min,60 min/次,5 d/周),普饮组和高脂组小鼠静置于没有开机的跑步机上,静置时间和频率与高脂运动组相同。HE染色观察各组小鼠肝组织病理学形态,Western Blotting法检测肝组织CNPY2、PERK、p-eIF2α蛋白表达。另培养人肝癌细胞系HepG2细胞,使用棕榈酸孵育肝细胞,在诱导NAFLD体外模型后,采用腺苷磷酸活化蛋白激酶(AMPK)激动剂模拟细胞水平运动效应,通过AMPK激动剂、CNPY2重组蛋白和(或)PERK抑制剂干预,Western Blotting检测各组肝细胞CNPY2、PERK、peIF2α、血红素加氧酶-1(HO-1)、核因子E2相关因子2(Nrf2)、NADPH氧化酶-1(NOX-1)蛋白表达、qRT-PCR检测各组肝细胞CNPY2 mRNA和PERK mRNA表达,ELISA检测各组肝细胞上清液白介素-1α(IL-1α)、白介素-6(IL-6)和肿瘤坏死因子(TNF-α)浓度。结果:(1)与普饮组比较,高脂组小鼠肝脏有明显脂肪性变,肝组织CNPY2、PERK、p-eIF2α蛋白表达显著升高(P<0.05);与高脂组比较,高脂运动组小鼠肝脏脂肪性变显著改善,肝组织CNP Y2、PERK、p-eIF2α蛋白表达显著降低(P<0.05)。(2)棕榈酸诱导肝细胞CNPY2、PERK、p-eIF2α、HO-1、Nrf2、NOX-1蛋白表达、CNPY2 mRNA、PERK mRNA表达和肝细胞上清液IL-1 α、IL-6、TNF-α浓度显著升高;CNPY2重组蛋白干预显著上调肝细胞以上指标水平,加重棕榈酸作用效应;AMPK激动剂和(或)PERK抑制剂ObjectiveTo explore the possible mechanism of aerobic exercise improving non-alcoholic fatty liver(NAFLD) by regulating endoplasmic reticulum stress protein kinase R-like ER kinase(PERK)/eukaryotic translation initiation factor 2α(eIF2α) pathway via canopy fibroblast growth factor signaling regulator 2(CNPY2).MethodsThirty eight-week male C57BL/6J mice were randomly divided into a normal group,a high-fat diet group and a high-fat diet + exercise group,each group of 10. The normal group was fed with normal diet,while the high-fat diet group and high-fat diet + exercise group were on high fat diet for 18 weeks until the end of the experiment. From the 10thweek,the mice in the high-fat diet + exercise group underwent daily 60-minute aerobic treadmill training five days a week at a speed of 12 m/min,while the other two groups were given fake exercise training. The pathological morphology of liver tissues was observed using the hematoxylin and eosin staining,while the expressions of CNPY2,PERK and phosphorylated eukaryotic translation initiation factor 2α(p-eIF2α) protein in the liver tissues were evaluated using Western blotting. Moreover, the human hepatoma cell line HepG2 was cultured,and hepatocytes were incubated with palmitic acid to induce NAFLD model in vitro. Then,the adenosine monophosphate-activated protein kinase(AMPK) agonist was used to simulate cellular movement,and AMPK agonist,CNPY2 recombinant protein and/or PERK inhibitor were employed to intervene. The protein expressions of CNPY2,PERK,p-eIF2α,haem oxygenase-1(HO-1), nuclear factor E2-related factor 2(Nrf2) and NADPH oxidase 1(NOX-1) were measured using Western blotting,while the interleukin-1 α(IL-1α),interleukin-6(IL-6) and tumor necrosis factor-a(TNF-α) in the hepatocellular supernatant were detected.ResultsCompared with the normal group,significant fatty degeneration, as well as increased CNPY2, PERK and p-eIF2α protein expression was observed in the high-fat diet group(P<0.05). Compared with the high-fat diet group,the expressions of

关 键 词：CNPY2重组蛋白 AMPK激动剂 PERK抑制剂 有氧运动 非酒精性脂肪肝 

分 类 号：G804.2[文化科学—运动人体科学] R575.5[文化科学—体育学]