CAJ | 학술논문

目的探讨白细胞介素-10(IL-10)对细菌脂蛋白(BLP)诱导大鼠乳鼠心肌细胞凋亡的抑制作用.方法 (1)体外培养大鼠乳鼠心肌细胞,以1μg/ml的BLP分别刺激心肌细胞0、12、24、36、48h.(2)以不同浓度的BLP(1 μg/ml和5μg/ml)和TNF-α(20 ng/ml和60 ng/ml)分别刺激心肌细胞24h.(3)将细胞分为五组,正常对照组、BLP(1μg/ml)组、IL-10(20 ng/ml)组、IL-10(20 ng/ml)+ BLP(1μg/ml)组及BLP(1μg/ml)+IL-10(20ng/ml)组,刺激细胞24h.提取细胞蛋白,Western blot法检测心肌细胞procaspase-3、cleaved caspase-3、bcl-2及bax蛋白的表达水平.结果 (1)BLP刺激心肌细胞24h后,caspase-3被明显激活,表现为cleaved caspase-3的表达逐渐增加.(2)与正常对照组比较,不同浓度的BLP和TNF-α分别刺激心肌细胞24h后可明显促进cleaved caspase-3表达增加(P＜0.01),且5μg/ml的BLP较1μg/ml的BLP刺激细胞后cleaved caspase-3的表达更高(P＜0.05).(3)与正常对照组相比,BLP组中心肌细胞cleaved caspase-3的表达明显升高(P＜0.01),而bcl-2/bax的比率显著降低(P＜0.01).与BLP组相比,IL-10+ BLP组和BLP+ IL-10组中,心肌细胞cleaved caspase-3水平显著降低(P＜0.01),而bcl-2/bax比率明显升高(P＜0.05),但这两组间无统计学差异.结论 IL-10对细菌脂蛋白诱导的乳大鼠心肌细胞凋亡具有抑制作用.
목적 탐토백세포개소-10(IL-10)대세균지단백(BLP)유도대서유서심기세포조망적억제작용.방법 (1)체외배양대서유서심기세포,이1μg/ml적BLP분별자격심기세포0、12、24、36、48h.(2)이불동농도적BLP(1 μg/ml화5μg/ml)화TNF-α(20 ng/ml화60 ng/ml)분별자격심기세포24h.(3)장세포분위오조,정상대조조、BLP(1μg/ml)조、IL-10(20 ng/ml)조、IL-10(20 ng/ml)+ BLP(1μg/ml)조급BLP(1μg/ml)+IL-10(20ng/ml)조,자격세포24h.제취세포단백,Western blot법검측심기세포procaspase-3、cleaved caspase-3、bcl-2급bax단백적표체수평.결과 (1)BLP자격심기세포24h후,caspase-3피명현격활,표현위cleaved caspase-3적표체축점증가.(2)여정상대조조비교,불동농도적BLP화TNF-α분별자격심기세포24h후가명현촉진cleaved caspase-3표체증가(P＜0.01),차5μg/ml적BLP교1μg/ml적BLP자격세포후cleaved caspase-3적표체경고(P＜0.05).(3)여정상대조조상비,BLP조중심기세포cleaved caspase-3적표체명현승고(P＜0.01),이bcl-2/bax적비솔현저강저(P＜0.01).여BLP조상비,IL-10+ BLP조화BLP+ IL-10조중,심기세포cleaved caspase-3수평현저강저(P＜0.01),이bcl-2/bax비솔명현승고(P＜0.05),단저량조간무통계학차이.결론 IL-10대세균지단백유도적유대서심기세포조망구유억제작용.
Objective To investigate the protective effects of interleukin-10 (IL-10) against bacteria lipoprotein( BLP)-induced apoptosis in neonatal rat cardiomyocytes.Methods Cardiomyocytes of neonatal rats were cultured in vitro and stimulated with 1 μg/ml BLP for 0,12,24,36 and 48 h.Cells were treated with different concentrations of BLP ( 1 μg/ml,5 μg/ml ) and TNF-α ( 20 ng/ml,60 ng/ml ) respectively for 24 h.In separate experiments,cells were divided into five groups:control groups,BLP( 1 μg/ml) groups,IL-10 ( 20 ng/ml) groups,IL-10(20 ng/ml) +BLP(1 μg/ml)groups,BLP(1 μg/ml) +IL-10(20 ng/ml) groups,and then treated for 24 h.The expression of procaspase-3,cleaved caspase-3,bcl-2 and bax were determined by Western blot.Results After stimulating the cardiomyocytes with 1 μg/ml BLP for 24 h,caspase-3 was notably activated,which was manifested by significantly increased cleaved caspase-3.Different concentrations of BLP and TNF-α both caused an apparent increase of cleaved caspase-3 ( P ＜ 0.01 ).Moreover,5 μg/ml BLP caused a higher increase of cleaved caspase-3 than 1 μg/ml BLP ( P ＜ 0.05 ).Compared with that in the control groups,BLP significantly up-regulated the expression of cleaved caspase-3 ( P ＜0.01 )and decreased the bcl-2/bax ratio( P ＜0.01 )in BLP groups.Compared with that in the BLP groups,in both IL-10 + BLP groups and BLP + IL-10 groups,cleaved caspase-3 was markedly down-regulated(P ＜ 0.01 ) and bcl-2/bax ratio was dramatically increased(P ＜ 0.05 ).But,there were no significant differences between these two groups.Conclusions IL-10 could protect cultured neonatal rat cardiomyocytes against bacteria lipoprotein-induced apoptosis.