硫化氢对apoE基因敲除小鼠肝脏内质网应激和脂质代谢的调节作用  

Regulatory effect of hydrogen sulfide on endoplasmic reticulum stress and lipid metabolism in livers in apoE knockout mice

作　　者：刘文莉[1] 陈志芳[1] 王海英[1] 涂家红[1] 关岚[1] 赵斌[1] 

机构地区：[1]北京积水潭医院急诊科,100035

出　　处：《中华老年医学杂志》2013年第12期1356-1360,共5页Chinese Journal of Geriatrics

摘　　要：目的探讨硫化氢对apoE基因敲除（apoE-/-）小鼠肝脏内质网应激（ERS）和脂质代谢的调节作用。方法雄性C57BL／6J和apoE-/-小鼠随机分为C57BL／6J对照组、apoE。组、apoE-/-硫氢化钠（NariS）组和apoE。＋炔丙基甘氨酸（PPG）组，均给予高脂饮食。对照组和apoE。组，每日腹腔注射生理盐水，apoE。＋NariS组每日腹腔注射硫化氢供体NariS，56／amol·kg-1·d，apoE-/-＋PPG组每日腹腔注射胱硫醚-Y-裂解酶抑制剂PPG，30mg·kg-1·d-1，10周后处死小鼠。检测血脂含量，采用油红（）染色检测肝脏脂质沉积，蛋白质印迹检测肝脏ERS相关蛋白葡萄糖调节蛋白78（GRP78）、磷酸化蛋白激酶R样内质网激酶（PPERK）、真核翻译起始因子一2d（elF2a）和胆固醇代谢相关蛋白胆固醇调节元件结合蛋白2（SREBP-2）、低密度脂蛋白（LDL）受体水平，实时定量PCR检测GRP78、SREBP-2基因的表达。结果与对照组比较，apoE-/-小鼠体质量无明显变化，但血浆中总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白（LDL）水平升高，肝脏脂质沉积增多，肝脏GRP78、PPERK和elF-2c蛋白表达显著升高；与apoE-/-小鼠比较，apoE-/- ＋NariS组小鼠血浆I。DL水平下降，肝脏脂质沉积改善，肝脏GRP78、p-PERK表达增多，p-elF-2ajelF2c比值增高，但肝脏SREBP-2、I。DI。受体的表达无明显变化。结论硫化氢可降低高脂饲养apoE。小鼠血浆I。DI．水平，改善肝脏脂肪沉积，上调GRP78蛋白及mRNA表达，促进PERK及elF2c~磷酸化，改善内质网功能，但对肝脏SREBP2、LDI，受体的表达无影响。Objective To investigate the regulatory effect of hydrogen sulfide （H^S） on endoplasmic reticulurn stress （ERS） and lipid metabolism in livers in apoE knockout （apo E / ） mice. Methods Male C57BI./6 J mice and homozygous apoE＇ mice were fed with a western type diet and randomly divided into four groups： C57BL/6 J control group （injected intraperitoneally with normal saline）, apoE＇ group （injected intraperitoneally with normal saline）, apoE / --NariS group （injected intraperitoneally with an H2S donor Naris 56#mol ~ kg ~ ~ d-t） and apoE/ ＋ DL - propargylgly- cine （PPG） group （injected intraperitoneally with an acystathionine y lyase inhibitor PPG 30 mg ~ kg ~ ~ d ~）. After 10 weeks, all mice were sacrificed and plasma lipids were detected. Lipid deposition was determined by oil red O staining. Glucose regulated protein78 （GRP78）, Thr-981 phosphorylated double stranded RNA activated protein kinase like endoplasmic reticulum kinase （PERK）, subunit of eukaryotic translation initiation factor-2a （elF-2a）, low density lipoprotein （LDL） receptor （LDLR）, sterol regulatory element-binding protein 2 （SREBP-2） in the livers were detected by Western bloting. The expressions of GRP78 and SREBP-2 mRNA were analyzed by real- time polymerase chain reaction （RT-PCR）. Results Compared with C57BL/6 J control group, plasma levels of total cholesterol （TC）, triglyeeride （TG） and low density lipoprotein （I.DL）, liver lipid content and expressions of GRP 78, PERK and eIF-2A were significantly increased in apoE mice, but body weight did not change. Compared with apoE/- mice, plasma LDL level was decreased, liver lipid deposition was improved, expressions of GRP-7g and PERK in ~ivers were increased, and the ratio of p elF-2a/ eIF-2a was increased in apoE j ~NaHS group, but expression levels of SREBP- 2 and I.DLR in liver did not change. Conclusions H2S decreases serum LDL level and liver lipid content, and up-regulates GRP78 protein and mRNA expressions, pro

关 键 词：硫化氢 小鼠 基因敲除 脂代谢障碍 

分 类 号：R575[医药卫生—消化系统]