CAJ | 학술논문

目的探讨盐酸戊乙奎醚(PHC)对脂多糖(LPS)诱导的内皮细胞损伤的影响及其作用机制.方法用RPMI 1640培养基将人脐静脉内皮细胞在5%CO2、37℃培养箱中培养,并分为对照组,LPS组及PHC组.检测细胞上清液乳酸脱氢酶(LDH)、细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)和一氧化氮(NO)水平.提取细胞核蛋白,Western blot分析细胞外信号调节激酶1/2(ERK1/2)和c-Jun氨基末端激酶(JNK)磷酸化蛋白的表达.采用SPSS 13.0统计软件进行分析,组间比较采用单因素方差分析,以P<0.05为差异具有统计学意义.结果与对照组比较.LPS组LDH含量[(1 642±367)U/L vs.(169±33)U/L]、MDA含量[(13.2±1.2)nmol/L vs.(7.2±0.8)nmoL/mL]及NO水平[(143.2±10.3)μmol/L vs.(85.5 4.4.1)μmol/L]明显增多,SOD活性明显下降[(41.2 ±2.7)U/mL vs.(61.1±2.8)U/mL],ERK1/2和JNK磷酸化表达明显增高.PHC可显著降低LDH含量[(392 4.90)U/L],降低MDA[(8.6±1.3)nmol/mL]和NO水平[(92.1 ±6.6)μmol/L],提高SOD活性[(58.0±3.0)U/mL],减弱ERK1/2磷酸化蛋白表达.结论盐酸戊乙奎醚对脂多糖诱导的内皮细胞损伤具有保护作用,其作用机制可能与抑制ERK1/2磷酸化表达减少过氧化物的产生有关.
목적 탐토염산무을규미(PHC)대지다당(LPS)유도적내피세포손상적영향급기작용궤제.방법 용RPMI 1640배양기장인제정맥내피세포재5%CO2、37℃배양상중배양,병분위대조조,LPS조급PHC조.검측세포상청액유산탈경매(LDH)、세포중병이철(MDA)、초양화물기화매(SOD)화일양화담(NO)수평.제취세포핵단백,Western blot분석세포외신호조절격매1/2(ERK1/2)화c-Jun안기말단격매(JNK)린산화단백적표체.채용SPSS 13.0통계연건진행분석,조간비교채용단인소방차분석,이P<0.05위차이구유통계학의의.결과 여대조조비교.LPS조LDH함량[(1 642±367)U/L vs.(169±33)U/L]、MDA함량[(13.2±1.2)nmol/L vs.(7.2±0.8)nmoL/mL]급NO수평[(143.2±10.3)μmol/L vs.(85.5 4.4.1)μmol/L]명현증다,SOD활성명현하강[(41.2 ±2.7)U/mL vs.(61.1±2.8)U/mL],ERK1/2화JNK린산화표체명현증고.PHC가현저강저LDH함량[(392 4.90)U/L],강저MDA[(8.6±1.3)nmol/mL]화NO수평[(92.1 ±6.6)μmol/L],제고SOD활성[(58.0±3.0)U/mL],감약ERK1/2린산화단백표체.결론 염산무을규미대지다당유도적내피세포손상구유보호작용,기작용궤제가능여억제ERK1/2린산화표체감소과양화물적산생유관.
Objective To investigate the effects of penehyclidine hydrochloride ( PHC) on lipopolysaccharide (LPS) -induced endothelial cells injury and its mechanism. Methods ECV-304 was cultured in RPMI1640 in a 5% humidified CO2 atmosphere at 37 ℃. Then cultured cells were used to assess the following treatments: control group, LPS group (1 μg/mL) and PHC group(2 μg/mL). At the end of the experiments, supernatant was collected for determination of lactate dehydrogenase ( LDH), and cells were collected for determination of malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO) levels. And extracellular regulated kinasel/2( ERKl/2)and JNK MAPK (mitogen-activated protein kinases, MAPK) protein expressions were determined using Western blot technique. Analysis of variance (ANOVA) was used for statistical analysis to compare values among all groups. A significant difference was presumed for a probability value < 0.05. Results Compared with control group, LDH leakage [(1642 ± 367) U/L vs (169±33)U/L], the contents of MDA[(13. 2 ± 1. 2) nmol/L vs (7. 2 ±0. 8)nmol/mL] and NO levels [(143.2 ± 10.3) μmol/L vs(85.5 ±4.1) μmol/L], expressions of ERK1/2 and JNK were remarkably increased and SOD activities[(41.2 ±2.7) U/mL vs (61. 1 ±2.8) U/mL] were obviously decreased in LPS group. PHC markedly decreased LDH leakage [(392 ±90) U/L], MDA contents [(8. 6 ± 1. 3) nmol/ mL] and NO levels [(92.1 ±6.6) μmol/L], ERK1/2 activation and enhanced SOD activities [(58.0± 3.0) U/mL]. Conclusions PHC could protect endothelial cells against LPS-induced cell injury. The effect of PHC is likely mediated through inhibition of ERK1/2 MAPK activation.