>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
PI3K/AKT/SIRT1信号通路介导H2S调节肝胆固醇代谢
作者:张睿1  马根山2  蔡君艳2 
单位:1. 东南大学 医学院, 江苏 南京 210009;
2. 东南大学附属中大医院 心血管内科, 江苏 南京 210009
关键词:硫化氢 PI3K/AKT SIRT1 HepG2细胞 胆固醇代谢 
分类号:R-33;R589.2
出版年·卷·期(页码):2019·38·第二期(230-237)
摘要:

目的:探讨硫化氢(H2S)上调SIRT1表达调控肝内胆固醇代谢过程,进而降低胆固醇水平的机制。方法:使用HepG2细胞建模,NaHS作为H2S供体,LY294002是PI3K/AKT通路抑制剂。利用不同浓度(0、50、100、200 μmol·L-1)NaHS处理HepG2细胞24 h,CCK8检测各组细胞活性,免疫印迹法检测各组SIRT1蛋白表达;100 μmol·L-1 NaHS分别处理细胞0、6、12、24 h,检测SIRT1蛋白表达;将细胞分成对照组、NaHS干预组及LY294002+NaHS干预组3组,免疫印迹法检测P-AKT、AKT和SIRT1,RT-qPCR检测SIRT1、SREBP-1c、SREBP-2、CYP7A1、HMGCR;将细胞分为空白对照组、油酸干预组(阳性对照)、油酸+NaHS组、油酸+NaHS+LY294002组,试剂盒检测细胞内胆固醇含量。结果:与对照组相比,50、100、200 μmol·L-1 NaHS对HepG2细胞活性无影响(P>0.05);与对照组相比,50、100、200 μmol·L-1 NaHS都能促进SIRT1蛋白表达并在100 μmol·L-1时达到最大(P<0.05);SIRT1蛋白表达呈时间依赖性升高,在24 h达到最高(P<0.05);与对照组相比,NaHS可以促进细胞内P-AKT/AKT和SIRT1蛋白表达,下调SREBP-1c、SREBP-2和HMGCR基因表达,上调CYP7A1基因表达,降低胞内胆固醇含量(P<0.05),联合应用LY294002具有相反的效应(P<0.05)。结论:PI3K/AKT/SIRT1通路参与了H2S对肝内胆固醇的调节作用。

Objective:To investigate the mechanism of H2S up-regulate SIRT1 to regulate the metabolism of cholesterol in the liver, thereby reducing the cholesterol level. Methods:HepG2 cells was used in the experiment. NaHS was exogenous H2S donor and LY294002 was the inhibitor of PI3K/AKT. HepG2 cells were cultivated with 4 different concentrations (0, 50, 100 and 200 μmol·L-1) of NaHS for 24 hours. Then the viability of HepG2 cells was detected by CCK-8 kits and the protein expressions of SIRT1were detected by western blot. Then 100 μmol·L-1 NaHS was selected to deal with HepG2 cells for 0 h, 6 h, 12 h, and 24 h respectively, western blot was used to detect SIRT1 protein expression. HepG2 cells were divided into blank control group, NaHS drug intervention group and LY294002+NaHS intervention group, then P-AKT, AKT and SIRT1 protein expressions were tested with western blot, gene expressions of SIRT1, SREBP-1c,SREBP-2,CYP7A1,HMGCR were detected with RT-qPCR. The cells were divided into blank control group, oleic acid intervention group (positive control),oleic acid+NaHS group and oleic acid+NaHS+LY294002 group, then intracellular cholesterol concentration were tested by using cholesterol testing kits. Results:Compared with the control group, 50,100,200 μmol·L-1 NaHS all had no effects on the viability of HepG2(P>0.05).The protein expressions of SIRT1 were increasing in a concentration and time dependence way and reached the maximum when the concentration was 100 μmol·L-1 and the time was 24 h(P<0.05). Compared with control group,H2S could increase P-AKT/AKT and SIRT1 protein expressions and CYP7A1 gene expression (P<0.05), inhibite SREBP-1c,SREBP-2 and HMGCR gene expressions (P<0.05), meanwhile, intracellular cholesterol levels reduced(P<0.05),using LY294002 processing cells in advance had the opposite effect. Conclusion:The PI3K/AKT/SIRT1 pathway is involved in the function of H2S in regulating cholesterol metabolism in the liver.

参考文献:

[1] WU D,ZHENG N,QI K,et al.Exogenous hydrogen sulfide mitigates the fatty liver in obese mice through improving lipid metabolism and antioxidant potential[J].Med Gas Res,2015,5(1):1.
[2] CHEUNG S H,KWOK W K,TO K F,et al.Anti-atherogenic effect of hydrogen sulfide by over-expression of cystathionine gamma-lyase (CSE) gene[J].PLoS One,2014,9(11):e113038.
[3] DU C,LIN X,XU W,et al.Sulfhydrated sirtuin-1 increasing its deacetylation activity is an essential epigenetics mechanism of anti-atherogenesis by hydrogen sulfide[J].Antioxid Redox Signal,2018,DOI:10.1089/ars.2017.7195.
[4] STEINL D C,KAUFMANN B A.Ultrasound imaging for risk assessment in atherosclerosis[J].Int J Mol Sci,2015,16(5):9749-9769.
[5] NOVAK J,BIENERTOVA-VASKU J,KARA T,et al.MicroRNAs involved in the lipid metabolism and their possible implications for atherosclerosis development and treatment[J].Mediators Inflamm,2014,DOI:10.1155/2014/275867
[6] AGABITI R E,SALVETTI M.Management of hypercholesterolemia, appropriateness of therapeutic approaches and new drugs in patients with high cardiovascular risk[J].High Blood Press Cardiovasc Prev,2016,23(3):217-230.
[7] KORBER M,KLEIN I,DAUM G.Steryl ester synthesis, storage and hydrolysis:a contribution to sterol homeostasis[J].Biochim Biophys Acta,2017,1862(12):1534-1545.
[8] SHARPE L J,HOWE V,PRABHU A V,et al.Navigating the shallows and rapids of cholesterol synthesis downstream of HMGCR[J]. J Nutr Sci Vitaminol (Tokyo),2015,61(Suppl):S154-S156.
[9] JESCH E D,CARR T P.Food ingredients that inhibit cholesterol absorption[J].Prev Nutr Food Sci,2017,22(2):67-80.
[10] LIU H,PATHAK P,BOEHME S,et al.Cholesterol 7alpha-hydroxylase protects the liver from inflammation and fibrosis by maintaining cholesterol homeostasis[J].J Lipid Res,2016,57(10):1831-1844.
[11] XIAO X,SONG B L.SREBP:a novel therapeutic target[J].Acta Biochim Biophys Sin,2013,45(1):2-10.
[12] LI T,KONG X,OWSLEY E,et al. Insulin regulation of cholesterol 7alpha-hydroxylase expression in human hepatocytes:roles of forkhead box O1 and sterol regulatory element-binding protein 1c[J].J Biol Chem,2006,281(39):28745-28754.
[13] BHATTACHARYA B S,SWEBY P K,MINIHANE A M,et al.A mathematical model of the sterol regulatory element binding protein 2 cholesterol biosynthesis pathway[J].J Theor Biol,2014,349:150-162.
[14] GRABOWSKA W,SIKORA E,BIELAK-ZMIJEWSKA A.Sirtuins, a promising target in slowing down the ageing process[J].Biogerontology,2017,18(4):447-476.
[15] KITADA M,OGURA Y,KOYA D.The protective role of Sirt1 in vascular tissue:its relationship to vascular aging and atherosclerosis[J].Aging (Albany NY),2016,8(10):2290-2307.
[16] SCHUG T T,LI X.Sirtuin 1 in lipid metabolism and obesity[J].Ann Med,2011,43(3):198-211.
[17] di MASI A, ASCENZI P.H2S:a "double face" molecule in health and disease[J].Biofactors,2013,39(2):186-196.
[18] FAES S, DORMOND O.PI3K and AKT:unfaithful partners in cancer[J].Int J Mol Sci,2015,16(9):21138-21152.
[19] KRYCER J R,SHARPE L J,LUU W,et al.The Akt-SREBP nexus:cell signaling meets lipid metabolism[J].Trends Endocrinol Metab,2010,21(5):268-276.
[20] 胡明珠,周波,盛琼,等.PI3K/Akt/Sirt1信号通路介导硫化氢后处理对大鼠缺血心肌的保护作用[J].中国药理学通报,2016(2):268-273.
[21] 廖玲.PCSK9在H_2S调节HepG2细胞脂质摄取中的作用及机制研究[D].衡阳:南华大学,2015.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 177341 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-87232481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364