中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
8期
836-839
,共4页
脂多糖%缺氧诱导因子-1α%心肌细胞%线粒体
脂多糖%缺氧誘導因子-1α%心肌細胞%線粒體
지다당%결양유도인자-1α%심기세포%선립체
Lipopolysaccharide%Hypoxia inducible factor-1α%Cardiomyocyte%Mitochondrial
目的 探讨心肌细胞缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)对脂多糖(lipopolysaccharide,LPS)诱导心肌细胞线粒体功能损伤的影响.方法 培养新生乳鼠心肌细胞,随机(随机数字法)分为4组.①C组:常规心肌细胞培养,不予任何处理；②LPS组:给予1μg/mlLPS,培养2h；③YC-1 +LPS组:预给HIF-1α特异性抑制剂YC-1 4μmol/L培养8h,再加入1 μg/mlLPS,培养2h；④YC-1组:仅给予4μmol/L YC-1培养8h.利用荧光分光光度计测定线粒体膜电位(mitochondrial membrane potential,MMP),荧光素酶法测定细胞ATP含量,比色法测定细胞色素C氧化酶(cytochrome c oxidase,COX)活性,Western blot测定细胞色素氧化酶亚单位1,2的蛋白表达水平.结果 与C组相比,LPS显著抑制心肌细胞MMP、COX活性,ATP含量也显著降低(P＜0.05),并抑制COX-1蛋白表达(P＜0.05),上调COX-2蛋白表达(P＜0.05).与LPS组相比,HIF-1α抑制剂YC-1能进一步降低心肌细胞MMP、COX活性及ATP含量(P＜0.01),显著增加COX-1表达并使COX-2表达的下调(P＜0.01).结论 HIF-1α抑制剂YC-1可能通过改变COX-1及COX-2表达,加重LPS所致的心肌细胞线粒体功能的损伤,提示HIF-1α对LPS诱导的心肌细胞线粒体损伤可能具有保护作用.
目的 探討心肌細胞缺氧誘導因子-1α(hypoxia-inducible factor-1α,HIF-1α)對脂多糖(lipopolysaccharide,LPS)誘導心肌細胞線粒體功能損傷的影響.方法 培養新生乳鼠心肌細胞,隨機(隨機數字法)分為4組.①C組:常規心肌細胞培養,不予任何處理；②LPS組:給予1μg/mlLPS,培養2h；③YC-1 +LPS組:預給HIF-1α特異性抑製劑YC-1 4μmol/L培養8h,再加入1 μg/mlLPS,培養2h；④YC-1組:僅給予4μmol/L YC-1培養8h.利用熒光分光光度計測定線粒體膜電位(mitochondrial membrane potential,MMP),熒光素酶法測定細胞ATP含量,比色法測定細胞色素C氧化酶(cytochrome c oxidase,COX)活性,Western blot測定細胞色素氧化酶亞單位1,2的蛋白錶達水平.結果 與C組相比,LPS顯著抑製心肌細胞MMP、COX活性,ATP含量也顯著降低(P＜0.05),併抑製COX-1蛋白錶達(P＜0.05),上調COX-2蛋白錶達(P＜0.05).與LPS組相比,HIF-1α抑製劑YC-1能進一步降低心肌細胞MMP、COX活性及ATP含量(P＜0.01),顯著增加COX-1錶達併使COX-2錶達的下調(P＜0.01).結論 HIF-1α抑製劑YC-1可能通過改變COX-1及COX-2錶達,加重LPS所緻的心肌細胞線粒體功能的損傷,提示HIF-1α對LPS誘導的心肌細胞線粒體損傷可能具有保護作用.
목적 탐토심기세포결양유도인자-1α(hypoxia-inducible factor-1α,HIF-1α)대지다당(lipopolysaccharide,LPS)유도심기세포선립체공능손상적영향.방법 배양신생유서심기세포,수궤(수궤수자법)분위4조.①C조:상규심기세포배양,불여임하처리；②LPS조:급여1μg/mlLPS,배양2h；③YC-1 +LPS조:예급HIF-1α특이성억제제YC-1 4μmol/L배양8h,재가입1 μg/mlLPS,배양2h；④YC-1조:부급여4μmol/L YC-1배양8h.이용형광분광광도계측정선립체막전위(mitochondrial membrane potential,MMP),형광소매법측정세포ATP함량,비색법측정세포색소C양화매(cytochrome c oxidase,COX)활성,Western blot측정세포색소양화매아단위1,2적단백표체수평.결과 여C조상비,LPS현저억제심기세포MMP、COX활성,ATP함량야현저강저(P＜0.05),병억제COX-1단백표체(P＜0.05),상조COX-2단백표체(P＜0.05).여LPS조상비,HIF-1α억제제YC-1능진일보강저심기세포MMP、COX활성급ATP함량(P＜0.01),현저증가COX-1표체병사COX-2표체적하조(P＜0.01).결론 HIF-1α억제제YC-1가능통과개변COX-1급COX-2표체,가중LPS소치적심기세포선립체공능적손상,제시HIF-1α대LPS유도적심기세포선립체손상가능구유보호작용.
Objective To investigate the effects of HIF-1α,on mitochondrial function of cardiomyocytes stimulated by lipopolysaccharide (LPS).Methods Cultured neonatal cardiomyocytes were randomly (random number) distributed to four groups (n =5).Control group:the neonatal cardiomyocytes were cultured in normal condition without any treatmeat; LPS group:cardiomyocytes were exposed to 1 μg/ml LPS for 2 h; YC-1 + LPS group:cardiomyocytes were exposed to 4 μmol/L YC-1 for 8 h before LPS treatment; YC-1 group:cardiomyocytes were exposed to 4μmol/L YC-1 for 8 h.As indexes of mitochondrial function,MMP,the content of ATP and the activity of cytochrome oxidase were measured,COX1/2 protein expression was determined by Western blotting.Results Compared with control group,LPS decreased MMP,the content of ATP and the activity of COX obviously ( P ＜0.05 ),depressed the expression of COX- 1 protein ( P ＜ 0.05 ),induced the expression of COX-2 protein ( P ＜ 0.05 ).Compared with the LPS group,MMP,the content of ATP and the activity of COX were markedly attenuated ( P ＜ 0.01 ),the expression of COX-1 was significantly increased and the expression of COX-2 was decreased in theYC-1 +LPS group (P ＜ 0.01 ).Conclusions YC-1 attributes to the dysfunction of mitochondrial of myocyte by alterating the expression of COX-1 and 2, HIF-1α may attenuate the mitochondrial dysfunction of cadiomyocytes stimulated by LPS.
