脂多糖对体外培养人脐静脉内皮细胞分泌功能及活力的影响

Effect of lipopolysaccharide on viability and secretion function of human umbilical vein endothelial cells

目的 :观察脂多糖对人脐静脉内皮细胞分泌缩血管因子内皮素 - 1、舒血管因子一氧化氮及其细胞活力的影响 ,为探讨感染性休克的发病机制提供实验资料。方法 :选用体外培养的 3代人脐静脉内皮细胞 ,分别使用浓度为 1g/L、10 0mg/L、10mg/L、1mg/L、10 0 μg/L、10 μg/L、1μg/L的脂多糖与之孵育 6h。分别使用放免法、硝酸还原酶法以及MTT法测定培养上清液内内皮素、一氧化氮的含量和内皮细胞的活力。结果 :正常对照组培养基中ET - 1的浓度 (pg/L)为 :2 5 1 6 4± 10 90。脂多糖与内皮细胞共孵育组培养基中ET - 1的浓度 (pg/L)分别为 :2 2 0 85± 19 14,2 78 6 7± 15 45 ,30 6 40± 11 6 0 ,312 87± 33 5 0 ,32 4 38± 17 0 2 ,2 91 49± 14 30 ,2 82 11± 13 38,各组与正常对照组比较 ,P <0 0 5或P <0 0 1;正常对照组培养基中总硝酸根离子 (NOx)的浓度 (μmol/L)为 6 2 9 46± 13 36 ,脂多糖与内皮细胞共孵育组培养基中总硝酸根离子 (NOx)的浓度 (μmol/L)分别为 :732 5 8± 2 3 2 1,6 6 9 87± 9 32 ,6 6 1 2 4± 16 80 ,6 5 0 33± 13 2 4,6 0 6 5 9± 12 94,6 2 6 75± 9 83,6 2 7 6 1± 5 6 1,各组与正常对照组比较 ,P <0 0 5或P <0 0 1。各组内皮细胞的活力分别为 :74%、81%。

AIM: To observe the direct effect of lipopolysaccharide (LPS) on secretion of endothelin-1(ET-1) and nitric oxide by human umbilical vein endothelial cell and cell viability of the secretor. METHODS: The third passage of human umbilical vein endothelial cells were incubated with different concentrations of LPS(1 g/L, 100 mg/L, 10 mg/L, 1 mg/L, 100 μg/L, 10 μg/L, 1 μg/L) for 6 hours, and the culture supernatants were collected. The concentrations of ET-1 were determined by radioimmunoassay, the concentrations of nitric oxide were determined using Greiss's method. The viabilities of cells were measured by MTT method. RESULTS: The concentration of ET-1 (pg/L) of normal control group was 251 64±10 90. The concentrations of ET-1(pg/L) of LPS treated groups were 220 85±19 14, 278 67±15 45, 306 40±11 60, 312 87±33 50, 324 38±17 02, 291 49±14 30, 282 11±13 38, respectively. (each group compared with normal control group, P<0.05 or P<0.01 ). The concentration of NO x (μmol/L) of normal control group was 629 46±13 36. The concentrations of NO x(μmol/L)of LPS treated groups were 732 58±23 21, 669 87±9 32,661 24±16 80, 650 33±13 24, 606 59±12 94, 626 75±9 83, 627 61±5 61,respectively(each group compared with normal control group, P<0 05 or P<0 01 ). The viabilities of endothelial cells of LPS treated groups were 74%, 81%, 86%, 88%,91%, 93%, 93%, respectively. CONCLUSION: LPS of lower concentrations had no significantly lethal effect on human umbilical vein endothelial cells, but enhanced secretion of ET-1 and inhibited NO production. LPS in higher concentrations showed significant lethal effect on human umbilical vein endothelial cells, inhibited secretion of ET-1 and enhanced NO production. [