CAJ | 학술논문

目的观察长托宁(PHC)对脂多精(LPS)引起的人脐静脉内皮细胞(HUVEC)损伤的保护作用,并探讨其机制.方法将体外培养HUVEC分为5组:空白对照组、LPS组、LPS/PHC( 10μg/L)组、LPS/PHC(25μ/L)组、LPS/PHC(50μg/L)组噻唑蓝(MTT)比色法测定内皮细胞的存活率:化学比色法测定乳酸脱氢酶(LDH)浓度；硝酸还原酶法测定一氧化氮(N0)浓度；定量逆转录-聚合酶链反应(RT-PCR)检测诱导型一氧化氮合酶(iNOS) mRNA表达；Western blot法检测iNOS蛋白质含量；酶联免疫吸附试验( ELISA)测定核因子-KB (NF-KB)和信号转导和转录激活因子3(STAT3)活性结果与空白对照组比较,LPS组细胞存活率[(60.31±8.76)％比(100.00±0.00)％]明显下降、LDH含量[(326±52) μmol/L比(125±25) μmol/L]和NO浓度[(55.49±10.16) μmol/L比(12.13±11.02) μmol/L]明显增加(P＜0.01)；而PHC可以逆转上述效应(P＜0.05).同时PHC可以降低LPS导致的内皮细胞iNOS转录和蛋白质表达水平,下调NF-KB和STAT3的表达(P＜0.05) 结论长托宁可以减轻LPS对内皮细胞的损伤,其作用可能是通过抑制NF-KB和STAT3信号系统,减少NO生成实现的.
목적 관찰장탁저(PHC)대지다정(LPS)인기적인제정맥내피세포(HUVEC)손상적보호작용,병탐토기궤제.방법 장체외배양HUVEC분위5조:공백대조조、LPS조、LPS/PHC( 10μg/L)조、LPS/PHC(25μ/L)조、LPS/PHC(50μg/L)조새서람(MTT)비색법측정내피세포적존활솔:화학비색법측정유산탈경매(LDH)농도；초산환원매법측정일양화담(N0)농도；정량역전록-취합매련반응(RT-PCR)검측유도형일양화담합매(iNOS) mRNA표체；Western blot법검측iNOS단백질함량；매련면역흡부시험( ELISA)측정핵인자-KB (NF-KB)화신호전도화전록격활인자3(STAT3)활성 결과 여공백대조조비교,LPS조세포존활솔[(60.31±8.76)％비(100.00±0.00)％]명현하강、LDH함량[(326±52) μmol/L비(125±25) μmol/L]화NO농도[(55.49±10.16) μmol/L비(12.13±11.02) μmol/L]명현증가(P＜0.01)；이PHC가이역전상술효응(P＜0.05).동시PHC가이강저LPS도치적내피세포iNOS전록화단백질표체수평,하조NF-KB화STAT3적표체(P＜0.05) 결론 장탁저가이감경LPS대내피세포적손상,기작용가능시통과억제NF-KB화STAT3신호계통,감소NO생성실현적.
Objective To investigate the protective effects of penehyclidine hvdrochloride (PHC)on lipopolysaccharide ( LPS)-induced human umbilical vein endothelial cells (HUVECs) injury and the underlying mechanism.Methods Cultured HUVECs were divided into five groups:control group,LPS group,LPS/PHC ( 10 μg/L) group,LPS/PHC (25 μg/L) group,LPS/PHC (50 μg/L) group.Cell viability was measured by methyl thiazol tetrazolium (MTT) colorimetric method.The concentrations of lactate dehydrogenase (LDH) and nitric oxide (NO) were measured respectively by using chemical colorimetric method and nitrate reduction method.The expression of inducible nitric oxide synthase (iNOS) mRNA and protein was detected hy using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting respectively.The activity of nuclear factor-KB (NF-KB) and signal transducer and activators of transcription 3 (STAT3) were measured by enzyme linked immunosorbent assay (ELISA).Results As compared with control group,cell viability [(60.31 ± 8.76 )％ vs 100.00％] was decreased and the concentrations of LDH [(326 ±52) μmol/L vs ( 125 ±25) μmoL/L] and NO [(55.49 ± 10.16) μmol/L vs ( 12.13 ±11.02) μmol/L] were increased in LPS group ( P ＜0.01 ) ; PHC could revere these effects on HUVECs (P ＜0.05).At the same time,PHC reduced the expression of iNOS mRNA and protein,and the activity of NF-KB and STAT3 ( P ＜ 0.05 ).Conclusion PHC can protect the endothelial function by reducing the concentration of NO.Inhibiting the NF-KB and STAT3 signal pathway may be the underlying mechanism.