中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2015年
5期
387-392
,共6页
孙艳%季文萱%单文红%刘桂美%黄俊彦
孫豔%季文萱%單文紅%劉桂美%黃俊彥
손염%계문훤%단문홍%류계미%황준언
造影剂%内质网应激%半胱氨酸天冬氨酸蛋白酶12%葡萄糖调节蛋白78%羟基酪醇
造影劑%內質網應激%半胱氨痠天鼕氨痠蛋白酶12%葡萄糖調節蛋白78%羥基酪醇
조영제%내질망응격%반광안산천동안산단백매12%포도당조절단백78%간기락순
Contrast media%Endoplasmic reticulum stress%Caspase 12%Glucoseregulated protein 78%Hydroxytyrosol
目的 研究造影剂肾病大鼠肾脏中葡萄糖调节蛋白78(GRP78)及半胱氨酸天冬氨酸蛋白酶12(Caspase-12)的表达情况,探讨内质网应激在造影剂肾病发病中的作用及羟基酪醇的干预作用.方法 84只大鼠随机分为3组:对照组(n=28)、模型组(n=28)和羟基酪醇干预组(n=28).分别于造模后12 h、24 h、48 h、72 h处死部分大鼠,检测各组大鼠血尿素氮(BUN)、血肌酐(Scr);ELISA法检测血清中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的含量;HE染色观察肾脏病理改变;原位末端转移酶标记法(TUNEL)检测肾小管上皮细胞凋亡;实时荧光定量PCR法检测肾组织GRP78 mRNA及Caspase-12mRNA的表达;免疫组化和Western印迹法检测各组大鼠肾组织GRP78及Caspase-12蛋白的表达.结果 对照组大鼠肾组织无明显病理损伤,GRP78、Caspase-12在mRNA水平及蛋白水平均无明显表达.与对照组相比,模型组大鼠肾组织病理损伤明显,细胞凋亡严重,且以上指标均显著升高(P＜0.05).羟基酪醇干预组大鼠肾组织病理损伤、细胞凋亡较模型组减轻,且GRP78、Caspase-12在mRNA水平及蛋白水平表达均较模型组明显降低(P＜0.05),但仍高于对照组(P＜0.05).结论 GRP78及Caspase-12介导的内质网应激可能参与大鼠造影剂肾病的发生发展;羟基酪醇在造影剂诱导的肾脏损伤中发挥保护作用,这可能与其减轻内质网应激有关.
目的 研究造影劑腎病大鼠腎髒中葡萄糖調節蛋白78(GRP78)及半胱氨痠天鼕氨痠蛋白酶12(Caspase-12)的錶達情況,探討內質網應激在造影劑腎病髮病中的作用及羥基酪醇的榦預作用.方法 84隻大鼠隨機分為3組:對照組(n=28)、模型組(n=28)和羥基酪醇榦預組(n=28).分彆于造模後12 h、24 h、48 h、72 h處死部分大鼠,檢測各組大鼠血尿素氮(BUN)、血肌酐(Scr);ELISA法檢測血清中丙二醛(MDA)、超氧化物歧化酶(SOD)、穀胱甘肽過氧化物酶(GSH-Px)的含量;HE染色觀察腎髒病理改變;原位末耑轉移酶標記法(TUNEL)檢測腎小管上皮細胞凋亡;實時熒光定量PCR法檢測腎組織GRP78 mRNA及Caspase-12mRNA的錶達;免疫組化和Western印跡法檢測各組大鼠腎組織GRP78及Caspase-12蛋白的錶達.結果 對照組大鼠腎組織無明顯病理損傷,GRP78、Caspase-12在mRNA水平及蛋白水平均無明顯錶達.與對照組相比,模型組大鼠腎組織病理損傷明顯,細胞凋亡嚴重,且以上指標均顯著升高(P＜0.05).羥基酪醇榦預組大鼠腎組織病理損傷、細胞凋亡較模型組減輕,且GRP78、Caspase-12在mRNA水平及蛋白水平錶達均較模型組明顯降低(P＜0.05),但仍高于對照組(P＜0.05).結論 GRP78及Caspase-12介導的內質網應激可能參與大鼠造影劑腎病的髮生髮展;羥基酪醇在造影劑誘導的腎髒損傷中髮揮保護作用,這可能與其減輕內質網應激有關.
목적 연구조영제신병대서신장중포도당조절단백78(GRP78)급반광안산천동안산단백매12(Caspase-12)적표체정황,탐토내질망응격재조영제신병발병중적작용급간기락순적간예작용.방법 84지대서수궤분위3조:대조조(n=28)、모형조(n=28)화간기락순간예조(n=28).분별우조모후12 h、24 h、48 h、72 h처사부분대서,검측각조대서혈뇨소담(BUN)、혈기항(Scr);ELISA법검측혈청중병이철(MDA)、초양화물기화매(SOD)、곡광감태과양화물매(GSH-Px)적함량;HE염색관찰신장병리개변;원위말단전이매표기법(TUNEL)검측신소관상피세포조망;실시형광정량PCR법검측신조직GRP78 mRNA급Caspase-12mRNA적표체;면역조화화Western인적법검측각조대서신조직GRP78급Caspase-12단백적표체.결과 대조조대서신조직무명현병리손상,GRP78、Caspase-12재mRNA수평급단백수평균무명현표체.여대조조상비,모형조대서신조직병리손상명현,세포조망엄중,차이상지표균현저승고(P＜0.05).간기락순간예조대서신조직병리손상、세포조망교모형조감경,차GRP78、Caspase-12재mRNA수평급단백수평표체균교모형조명현강저(P＜0.05),단잉고우대조조(P＜0.05).결론 GRP78급Caspase-12개도적내질망응격가능삼여대서조영제신병적발생발전;간기락순재조영제유도적신장손상중발휘보호작용,저가능여기감경내질망응격유관.
Objective To investigate the expression of glucose-regulated protein 78(GRP78)and cysteine aspartic acid protease 12(Caspase-12) and evaluate the endoplasmic reticulum stress (ERS) in rats with contrast-induced nephropathy (CIN),and observe the protective effects of hydroxytyrosol on CIN rats.Methods Eighty-four Wistar rats,(220±20) g,were randomly divided into control group,CIN group,hydroxytyrosol treated group (group C+H).At 12th,24th,48th,72th day after the rats model were established,BUN and Scr were detected.ELISA were used to detect the expression of methane dicarboxylic aldehyde (MDA),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px).HE staining were used to evaluate the pathological change of kidney.TUNEL were used to detect the apoptosis of tubular ceils.Real-time PCR were used to detect the expression of GRP78 mRNA and Caspase-12 mRNA in tubular cells.Immunohistochemistry and Western blotting were used to detect the expression of GRP78 and Caspase-12 protein in tubular cells.Results BUN,Scr,the mRNA and protein expression of GRP78,Caspase-12 in hydroxytyrosol treated group were higher than that in control group(P ＜ 0.05),but were significantly lower than that in CIN group (P ＜ 0.05).Pathological changes and the apoptosis of tubular cells in CIN group were more serious than that in hydroxytyrosol treated group (P ＜ 0.05).Conclusions Endoplasmic reticulum stress may be associated with contrast-induced nephropathy.Hydroxytyrosol can protect kidney from contrast medium via reducing the endoplasmic reticulum stress.
