中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2010年
4期
500-503
,共4页
蛋白质修饰,转译后%HSP70热休克蛋白质类%谷氨酰胺%内毒素类%肌细胞,心脏
蛋白質脩飾,轉譯後%HSP70熱休剋蛋白質類%穀氨酰胺%內毒素類%肌細胞,心髒
단백질수식,전역후%HSP70열휴극단백질류%곡안선알%내독소류%기세포,심장
Protein processing,post-translational%HSP70 heat-shock proteins%Gintamine%Endotoxins%Myocytes,cardiac
目的 探讨O位-N-乙酰葡萄糖胺(O-GlcNAc)修饰在谷氨酰胺(Gln)诱导LPS干预的大鼠心肌细胞热休克蛋白70(HSP70)表达中的作用.方法 出生48 h的SD大鼠,原代培养心肌细胞,随机分为6组,每组11瓶(1×106个,瓶):对照组(C组)加入双蒸水25 μl;Gln组加入Gln,终浓度5 mmol/L;LPS组加入LPS,终浓度4 μg/ml;Gin+LPS组依次加入Gln和LPS,Gin终浓度5 mmol/L,LPS 终浓度4 μg/ml;Gln+LPS+Alloxan组依次加入Gln、LPS和O位-N-乙酰氨基葡萄糖转移酶抑制剂Alloxan,Gln和LPS终浓度与Gln+LPS组相同,Alloxan终浓度1 mmol/L;Gln+LPS+PUGNAc组依次加入Gln、LPS和O位-N-乙酰氨基葡萄糖苷酶抑制剂PUGNAc,Gln和LPS终浓度与Gln+LPS组相同,PUGNAc终浓度100 μmol/L.各组孵育6 h后测定心肌细胞存活率、O-GlcNAc和HSP70的表达水平.结果 各组大鼠心肌细胞存活率比较差异无统计学意义(P>0.05);与C组比较,其余组心肌细胞0.GlcNAc和HSP70的表达上调(P<0.05);与LPS组比较,Gln+LPS组和Gln+LPS+PUGNAc组心肌细胞O-GlcNAc和HSP70的表达上调(P<0.05);与Gln+LPS组比较,Gin+LPS+Alloxan组心肌细胞0-GlcNAc和HSP70的表达下调,Gln+LPS+PUGNAc组心肌细胞O-GlcNAc和HSP70的表达上调(P<0.05).结论 Gln诱导LPS干预大鼠心肌细胞HSPT0表达上调的机制可能与O-GlcNAc修饰有关.
目的 探討O位-N-乙酰葡萄糖胺(O-GlcNAc)脩飾在穀氨酰胺(Gln)誘導LPS榦預的大鼠心肌細胞熱休剋蛋白70(HSP70)錶達中的作用.方法 齣生48 h的SD大鼠,原代培養心肌細胞,隨機分為6組,每組11瓶(1×106箇,瓶):對照組(C組)加入雙蒸水25 μl;Gln組加入Gln,終濃度5 mmol/L;LPS組加入LPS,終濃度4 μg/ml;Gin+LPS組依次加入Gln和LPS,Gin終濃度5 mmol/L,LPS 終濃度4 μg/ml;Gln+LPS+Alloxan組依次加入Gln、LPS和O位-N-乙酰氨基葡萄糖轉移酶抑製劑Alloxan,Gln和LPS終濃度與Gln+LPS組相同,Alloxan終濃度1 mmol/L;Gln+LPS+PUGNAc組依次加入Gln、LPS和O位-N-乙酰氨基葡萄糖苷酶抑製劑PUGNAc,Gln和LPS終濃度與Gln+LPS組相同,PUGNAc終濃度100 μmol/L.各組孵育6 h後測定心肌細胞存活率、O-GlcNAc和HSP70的錶達水平.結果 各組大鼠心肌細胞存活率比較差異無統計學意義(P>0.05);與C組比較,其餘組心肌細胞0.GlcNAc和HSP70的錶達上調(P<0.05);與LPS組比較,Gln+LPS組和Gln+LPS+PUGNAc組心肌細胞O-GlcNAc和HSP70的錶達上調(P<0.05);與Gln+LPS組比較,Gin+LPS+Alloxan組心肌細胞0-GlcNAc和HSP70的錶達下調,Gln+LPS+PUGNAc組心肌細胞O-GlcNAc和HSP70的錶達上調(P<0.05).結論 Gln誘導LPS榦預大鼠心肌細胞HSPT0錶達上調的機製可能與O-GlcNAc脩飾有關.
목적 탐토O위-N-을선포도당알(O-GlcNAc)수식재곡안선알(Gln)유도LPS간예적대서심기세포열휴극단백70(HSP70)표체중적작용.방법 출생48 h적SD대서,원대배양심기세포,수궤분위6조,매조11병(1×106개,병):대조조(C조)가입쌍증수25 μl;Gln조가입Gln,종농도5 mmol/L;LPS조가입LPS,종농도4 μg/ml;Gin+LPS조의차가입Gln화LPS,Gin종농도5 mmol/L,LPS 종농도4 μg/ml;Gln+LPS+Alloxan조의차가입Gln、LPS화O위-N-을선안기포도당전이매억제제Alloxan,Gln화LPS종농도여Gln+LPS조상동,Alloxan종농도1 mmol/L;Gln+LPS+PUGNAc조의차가입Gln、LPS화O위-N-을선안기포도당감매억제제PUGNAc,Gln화LPS종농도여Gln+LPS조상동,PUGNAc종농도100 μmol/L.각조부육6 h후측정심기세포존활솔、O-GlcNAc화HSP70적표체수평.결과 각조대서심기세포존활솔비교차이무통계학의의(P>0.05);여C조비교,기여조심기세포0.GlcNAc화HSP70적표체상조(P<0.05);여LPS조비교,Gln+LPS조화Gln+LPS+PUGNAc조심기세포O-GlcNAc화HSP70적표체상조(P<0.05);여Gln+LPS조비교,Gin+LPS+Alloxan조심기세포0-GlcNAc화HSP70적표체하조,Gln+LPS+PUGNAc조심기세포O-GlcNAc화HSP70적표체상조(P<0.05).결론 Gln유도LPS간예대서심기세포HSPT0표체상조적궤제가능여O-GlcNAc수식유관.
Objective To investigate the effects of O-GlcNAc modification on gintamine (Glu)-induced heat shock protein 70 (HSP70) expression in LPS-treated rat cardiomyocytes.Methods Primary cultures of neonatal rat cardiomyocytes were randomly divided into 6 groups:group Ⅰ control(group C);group Ⅱ Glu;group Ⅲ LPS;group Ⅳ Glu+LPS;group Ⅴ Glu+LPS+Alloxan and group Ⅵ Gln+LPS+PUGNAc.In group C double distilled water 25 μl was added.In group Ⅱ-Ⅵ the cells were exposed to the sanle concentrations of Glu (5 mmol/L)and LPS(4 μg/ml) except Alloxan (an inhibiter of O-linked β-N-acetyl glucosamine transferase/OGT) (1 mmol/L) and PUGNAc (an inhibitor of O-linked β-N-acetyl glucosaminidase/OGA)(100μmol/L).After being incubated for 6 h,cardiomyocyte viability,O-GlcNAc modification level and HSP70 expression level were measured.Results There was no significant difference in cell viability among the six groups.The levels of O-GlcNAc modification and HSP70 expression were significantly higher in group Ⅱ-Ⅵ than in group Ⅰ,were significantly higher in group Ⅳ and group Ⅵ than in group Ⅲ,were significantly lower in group Ⅴ and higher in group Ⅵ than in group Ⅳ.Conclusion O-GlcNAc modification may be involved in Glu-induced HSPT0 expression in LPS-treated cardiomyocytes.
