CAJ | 학술논문

目的研究舍曲林对对NGF诱导的PC12细胞细胞活力以及酪氨酸羟化酶(TH)和磷酸化细胞外信号调节激酶1/2(pERK1/2)表达的影响.方法以NGF诱导后的PC12细胞为细胞模型,给予5μM、10μM、20μM、50μM不同剂量舍曲林,分别进行直接作用和保护作用处理后,采用细胞计数试剂盒-8(CCK-8)检测细胞活性,免疫组织化学观察细胞形态学改变以及蛋白质斑迹法(western blot)检测PC12细胞酪氨酸羟化酶(TH)和磷酸化细胞外信号调节激酶(pERK1/2)的表达水平的变化.结果中等剂量的舍曲林促进PC12细胞的活性,直接作用24h后20μM(1.32±0.11)、10μM(1.17±0.05),保护作用24h后20μM(1.15±0.11)显著高于对照组,高剂量50μM的舍曲林PC12细胞有很强的毒性作用.20μM舍曲林直接作用后PCI2细胞TH表达增加,TH与β-actin表达阳性面积的比值24h作用组(1.27±0.05)以及48 h作用组(1.23±0.08)与对照组(作用24h和48 h分别为0.99±0.04,0.92±0.07)相比差异有统计学意义(P＜0.01,P＜0.05),20μM 舍曲林直接作用后PC12细胞pERK1/2表达也增加,pERK1/2与β-actin表达阳性面积的比值24 h作用组(1.41±0.05)以及48 h作用组(1.40±0.06)与对照组(作用24 h和48 h分别为0.86±0.04,0.78±0.06)相比差异有统计学意义(P＜0.01,P＜0.05),免疫组织化学结果与上述一致.结论舍曲林能提高PC12细胞的活性,对神经元提供保护可能是舍曲林抗抑郁作用机制之一,而这种作用可能是通过上调TH以及pERK1/2的表达实现的.
목적 연구사곡림대대NGF유도적PC12세포세포활력이급락안산간화매(TH)화린산화세포외신호조절격매1/2(pERK1/2)표체적영향.방법 이NGF유도후적PC12세포위세포모형,급여5μM、10μM、20μM、50μM불동제량사곡림,분별진행직접작용화보호작용처리후,채용세포계수시제합-8(CCK-8)검측세포활성,면역조직화학관찰세포형태학개변이급단백질반적법(western blot)검측PC12세포락안산간화매(TH)화린산화세포외신호조절격매(pERK1/2)적표체수평적변화.결과 중등제량적사곡림촉진PC12세포적활성,직접작용24h후20μM(1.32±0.11)、10μM(1.17±0.05),보호작용24h후20μM(1.15±0.11)현저고우대조조,고제량50μM적사곡림PC12세포유흔강적독성작용.20μM사곡림직접작용후PCI2세포TH표체증가,TH여β-actin표체양성면적적비치24h작용조(1.27±0.05)이급48 h작용조(1.23±0.08)여대조조(작용24h화48 h분별위0.99±0.04,0.92±0.07)상비차이유통계학의의(P＜0.01,P＜0.05),20μM 사곡림직접작용후PC12세포pERK1/2표체야증가,pERK1/2여β-actin표체양성면적적비치24 h작용조(1.41±0.05)이급48 h작용조(1.40±0.06)여대조조(작용24 h화48 h분별위0.86±0.04,0.78±0.06)상비차이유통계학의의(P＜0.01,P＜0.05),면역조직화학결과여상술일치.결론 사곡림능제고PC12세포적활성,대신경원제공보호가능시사곡림항억욱작용궤제지일,이저충작용가능시통과상조TH이급pERK1/2적표체실현적.
Objective To investigate the effect of sertraline on the viability and the expression of tyrosine hydroxylase (TH) and phosphorylated ERK1/2 in NGF-induced rat pheochromocytoma (PC12) cells.Methods NGF-induced PC12 cells were pretreated or directly treated with different concentrations of sertraline for 24 or 48 hours and the pretreated groups were then subjected to serum withdrawal condition. Then cell viability was determined by the cell counting Kit-8 (CCK-8). The expression of Tyrosine hydroxylase (TH) and pERK1/2in NGF-induced PC12 cells was determined by immunohistochemistry and western blot respectively. Results The viability of NGF-induced PC12 was improved after administration with sertra]ine. After 24h sertraline administration, the cells activity of PC 12 cells at 20μM ( 1.32 ± 0. 11 ) , 10μM ( 1. 17 ± 0.05 ) of direct effect, and 20μM ( 1.15 ±0.11 ) of protect effect increased dramatically as compared with control group. But high dose ( 50μM )sertraline express high toxic effect to PC12 cells. The expression of TH was increased by sertraline 20 μM at both 24h(ratio of TH/β-actin = 1.27 ±0.05) and 48h(ratio of TH/β-actin = 1. 23 ±0.08) compare with control group,and the expression of pERK1/2 also increased dramatically by sertraline 20 μM at both 24h (ratio of (pERK1/2)/β-actin = 1.41±0.05) and 48h( ratio of (pERK1/2)/β-actin = 1.40 ±0.06) compare with control group(P＜0. 01, P ＜ 0. 05). Immunohistochemistry showed similar results. Conclusion These data suggest that the neuroprotective effect of sertraline may play an important role in depression therapy, and this effect might be mediated by TH and pERK1/2 up-regulation.