L-NAME对内毒素致大鼠肺动脉内皮细胞损伤的保护作用

Protection of L-NAME on RPAECs damage induced by LPS

目的探讨左旋硝基精氨酸甲酯(L-NAME)是否对内毒素脂多糖(LPS)导致的大鼠肺动脉内皮细胞(RPAECs)损伤具有保护作用。方法利用大鼠肺动脉组织块贴壁法进行细胞原代培养。通过形态观察和Ⅷ因子相关抗原抗血清间接免疫荧光染色对细胞进行鉴定。将细胞分组,LPS、L-NAME与细胞共育24小时,检测各组细胞及培养上清中的硝基酪氨酸阳性细胞百分比(NT%)和乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、内皮素-1(ET-1)的含量。结果L-NAME组中LDH(2237.04±50.32,2104.26±58.57)、MDA(8.36±0.25,7.26±0.24)比LPS组LDH(2553.01±55.75)、MDA(10.95±0.33)明显降低(P<0.05),SOD(41.09±2.20,50.76±2.66)较LPS组SOD(33.16±2.9)明显升高(P<0.05),NT%(33.7±1.94%,28.88±1.73%)与LPS组(40.28±2.3%)相比显著下降(P<0.05),ET-1为(417.98±21.44,467.8±17.94 pg/m l,505.88±20 pg/m l)与LPS组ET-1(391.67±14.62 pg/m l)相比明显升高(P<0.05)。结论L-NAME可能通过抗氧化作用来保护大鼠肺动脉内皮细胞。但是L-NAME升高细胞培养上清中的ET-1,单独用药可能导致肺动脉压力的增高。

Objective： To explore the protection of L-NAME in the RPAECS damage induced by LPS. Methods： ① Pulmonary artery endothelial cells were isolated and cultured by explant technique. ②The cells were identified through the observation of antigen anti - serum associated with Ⅷ factor by indirect immunofluorescence staining. ③The cells were randomly divided into three groups： the control group, the LPS group and the L-NAME group and cultured with LPS and L- NAME for 24 hours. ④ For the index detection, LDH, MDA, SOD, ET-1 and NT masculine percentage（NT% ） of every group＇s supernatant or cells were included. Results： Compared with the LPS group, the L-NAME group had lower LDH （2237.04 ± 50.32,2104.26±58.57 ）, MDA （ 8.36 ± 0.25,7.26 ± 0.24 ）, NT% （ 33.7 ±1.94%, 28.88 ± 1.73 % ）, ET-1 （417.98 ±21.44,467.8 ± 17.94 pg/ml,505.88 ±20 pg/ml） （P 〈0.05 ） and higher SOD（41.09 ±2.20,50.76 ±2.66 ） （P 〈 0. 05 ）. Conclusion ： L-NAME can lessen PAECs damage caused by LPS, which can be seen from the changes of LDH, MDA, SOD and NT%. However, ET-1 with the L-NAME rise in cell culture medium may lead to increased pul- monary artery pressur