中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2010年
3期
387-391
,共5页
心肌肥厚%肿瘤坏死因子-α%钙离子%钙调素依赖蛋白激酶%信号转导%KN93
心肌肥厚%腫瘤壞死因子-α%鈣離子%鈣調素依賴蛋白激酶%信號轉導%KN93
심기비후%종류배사인자-α%개리자%개조소의뢰단백격매%신호전도%KN93
cardiomyocyte hypertrophy%tumor necrosis factor-α%calcium%calmodulin-dependent kinase%signal transduction%KN93
目的 研究钙调素依赖蛋白激酶Ⅱ(CaMKⅡ)信号通路在肿瘤坏死因子-α(TNF-α)诱导心肌细胞肥大中的作用.方法 Lowry法测心肌细胞蛋白含量;计算机图像分析系统测心肌细胞体积;[~3H]-亮氨酸参入法测心肌细胞蛋白合成;Till阳离子测定系统观察胞内[Ca~(2+)]_i瞬变;Western blot法测定CaMKⅡδ_B的表达.结果 ① TNF-α(100 μg·L~(-1))明显诱导心肌细胞蛋白含量、蛋白合成及体积的增加,CaMKⅡ特异性抑制剂KN93(0.2 μmol·L~(-1))明显抑制TNF-α诱导的心肌肥大,但对正常心肌细胞生长无影响.② TNF-α引起心肌细胞内钙离子浓度([Ca~(2+)]_i)瞬间变化幅度增高,KN93明显降低TNF-α诱导的上述改变.③ TNF-α明显增强心肌细胞内CaMKⅡδ_B的表达.结论 TNF-α可能通过引起心肌细胞[Ca~(2+)]_i升高,促进CaMKⅡδ_B表达诱导心肌细胞肥大的.
目的 研究鈣調素依賴蛋白激酶Ⅱ(CaMKⅡ)信號通路在腫瘤壞死因子-α(TNF-α)誘導心肌細胞肥大中的作用.方法 Lowry法測心肌細胞蛋白含量;計算機圖像分析繫統測心肌細胞體積;[~3H]-亮氨痠參入法測心肌細胞蛋白閤成;Till暘離子測定繫統觀察胞內[Ca~(2+)]_i瞬變;Western blot法測定CaMKⅡδ_B的錶達.結果 ① TNF-α(100 μg·L~(-1))明顯誘導心肌細胞蛋白含量、蛋白閤成及體積的增加,CaMKⅡ特異性抑製劑KN93(0.2 μmol·L~(-1))明顯抑製TNF-α誘導的心肌肥大,但對正常心肌細胞生長無影響.② TNF-α引起心肌細胞內鈣離子濃度([Ca~(2+)]_i)瞬間變化幅度增高,KN93明顯降低TNF-α誘導的上述改變.③ TNF-α明顯增彊心肌細胞內CaMKⅡδ_B的錶達.結論 TNF-α可能通過引起心肌細胞[Ca~(2+)]_i升高,促進CaMKⅡδ_B錶達誘導心肌細胞肥大的.
목적 연구개조소의뢰단백격매Ⅱ(CaMKⅡ)신호통로재종류배사인자-α(TNF-α)유도심기세포비대중적작용.방법 Lowry법측심기세포단백함량;계산궤도상분석계통측심기세포체적;[~3H]-량안산삼입법측심기세포단백합성;Till양리자측정계통관찰포내[Ca~(2+)]_i순변;Western blot법측정CaMKⅡδ_B적표체.결과 ① TNF-α(100 μg·L~(-1))명현유도심기세포단백함량、단백합성급체적적증가,CaMKⅡ특이성억제제KN93(0.2 μmol·L~(-1))명현억제TNF-α유도적심기비대,단대정상심기세포생장무영향.② TNF-α인기심기세포내개리자농도([Ca~(2+)]_i)순간변화폭도증고,KN93명현강저TNF-α유도적상술개변.③ TNF-α명현증강심기세포내CaMKⅡδ_B적표체.결론 TNF-α가능통과인기심기세포[Ca~(2+)]_i승고,촉진CaMKⅡδ_B표체유도심기세포비대적.
Aim To investigate whether Ca~(2+)/calmodulin-dependent kinase Ⅱ(CaMKⅡ)contribute to tumor necrosis factor α(TNF-α)-induced cardiomyocyte hypertrophy.Methods The protein content was assayed with Lowry's method.The cardiomyocytes volumes were measured by computer photograph analysis system.The protein synthesis was assayed with[~3H]-lencine incorporation method.[Ca~(2+)]_i transient was measured by Till image system by cell-loading Fura-2/AM.The expression of CaMKⅡδ_B was determined by Western blot.Results ① TNF-α significantly induced the increase of protein content, [~3H]-leucine incorporation and cell size;These responses were significantly suppressed by KN93, a selective CaMKⅡ inhibitor.② TNF-α increased the amplitude of the spontaneous Ca~(2+) transients in cultured ventricular myocytes from the neonatal rat;CaMKⅡ inhibitor KN93 can suppress the elevation induced by TNF-α.③ TNF-α significantly increased the expression of CaMKⅡδ_B.Concluslon CaMKⅡ signal pathway are involved in TNF-α-induced cardiomyocyte hypertrophy in rats.
