CAJ | 학술논문

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/6 J和apoE小鼠随机分为C57BL/6 J对照组、apoE-/-组、apoE-/-+硫氢化钠(NaHS)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-/-+NaHS组每日腹腔注射硫化氢供体NaHS,56μmol·kg-1·d-1,apoE-/-十PPG组每日腹腔注射胱硫醚-γ-裂解酶抑制剂PPG,30 mg· kg-1·d-1,10周后处死小鼠.检测血脂含量,采用油红O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(p PERK)、真核翻译起始因子-2α(eIF-2α)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、p-PERK和eIF-2α蛋白表达显著升高;与apoE-/-小鼠比较,apoE-/-+NaHS组小鼠血浆LDL水平下降,肝脏脂质沉积改善,肝脏GRP78、p-PERK表达增多,p-eIF-2α/ eIF-2α比值增高,但肝脏SREBP 2、LDL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LDL水平,改善肝脏脂肪沉积,上调GRP78蛋白及mRNA表达,促进PERK及eIF-2α磷酸化,改善内质网功能,但对肝脏SREBP-2、LDL受体的表达无影响.
목적 탐토류화경대apoE기인고제(apoE-/-)소서간장내질망응격(ERS)화지질대사적조절작용. 방법 웅성C57BL/6 J화apoE소서수궤분위C57BL/6 J대조조、apoE-/-조、apoE-/-+류경화납(NaHS)조화apoE-/-+결병기감안산(PPG)조,균급여고지음식.대조조화apoE-/-조,매일복강주사생리염수,apoE-/-+NaHS조매일복강주사류화경공체NaHS,56μmol·kg-1·d-1,apoE-/-십PPG조매일복강주사광류미-γ-렬해매억제제PPG,30 mg· kg-1·d-1,10주후처사소서.검측혈지함량,채용유홍O염색검측간장지질침적,단백질인적검측간장ERS상관단백포도당조절단백78(GRP78)、린산화단백격매R양내질망격매(p PERK)、진핵번역기시인자-2α(eIF-2α)화담고순대사상관단백담고순조절원건결합단백2(SREBP-2)、저밀도지단백(LDL)수체수평,실시정량PCR검측GRP78、SREBP-2기인적표체. 결과 여대조조비교,apoE-/-소서체질량무명현변화,단혈장중총담고순(TC)、삼선감유(TG)、저밀도지단백(LDL)수평승고,간장지질침적증다,간장GRP78、p-PERK화eIF-2α단백표체현저승고;여apoE-/-소서비교,apoE-/-+NaHS조소서혈장LDL수평하강,간장지질침적개선,간장GRP78、p-PERK표체증다,p-eIF-2α/ eIF-2α비치증고,단간장SREBP 2、LDL수체적표체무명현변화. 결론 류화경가강저고지사양apoE-/-소서혈장LDL수평,개선간장지방침적,상조GRP78단백급mRNA표체,촉진PERK급eIF-2α린산화,개선내질망공능,단대간장SREBP-2、LDL수체적표체무영향.
Objective To investigate the regulatory effect of hydrogen sulfide (H2S) on endoplasmic reticulum stress (ERS) and lipid metabolism in livers in apoE knockout (apo E-/-) mice.Methods Male C57BL/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-/-+NaHS group (injected intraperitoneally with an H2S donor NaHS 56μmol · kg-1 · d-1) and apoE-/-+ DL-propargylglycine (PPG) group (injected intraperitoneally with an acystathionine-γ-lyase inhibitor PPG 30 mg ·kg-1 · d-1).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-2α (elF-2α),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 realtime polymerase chain reaction (RT-PCR).Results Compared with C57BL/6 J control group,plasma levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein (LDL),liver lipid content and expressions of GRP-78,PERK and eIF-2α 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-78 and PERK in livers were increased,and the ratio of p-eIF-2α/ eIF-2α was increased in apoE-/-+NaHS group,but expression levels of SREBP-2 and LDLR in liver did not change.Conclusions H2S decreases serum LDL level and liver lipid content,and up regulates GRP78 protein and mRNA expressions,promotes PERK and eIF2α phosphorylations,improves endoplasmic reticulum function,but has no effect on the expressions of SREBP-2 and LDLR in apoE-/-mice fed with a high-fat diet.