异丙酚对内毒素诱导人脐静脉内皮细胞过氧亚硝基阴离子生成的影响

目的 探讨异丙酚对内毒素诱导人脐静脉内皮细胞过氧亚硝基阴离子(ONOO-)生成的影响.方法 培养至活细胞计数大于95%的人脐静脉内皮细胞,随机分为7组(n=5),对照组(C组)不给予任何处理;LOS0.1组、LPS1组和LPS10组分别加入内毒素(LPS)至终浓度为0.1、1和10 μg/ml,于37℃5%CO2培养箱中孵育6 h;P4+LPS10组和P40+LPS10组预先加入异丙酚至终浓度为4、40μg/ml,I40+LPS10组预先加入脂质溶剂Introlipid至终浓度为40 μg/ml,于37℃ 5%CO2培养箱中孵育30 min,再分别加入LPS至终浓度为10μg/ml,于培养箱中继续孵育6 h.孵育6 h时,测定细胞活力和乳酸脱氢酶(LDH)释放率;采用免疫组化法和Western blot法测定硝基酪氨酸蛋白(NT)表达.结果 与C组比较,其余各组细胞活力降低,内皮细胞NT表达上调,LPS1组、LPS10组、I40+LPS10组、P4+LPS10组和P40+LPS10组LDH释放率升高(P<0.01);与LPS0.1组比较,LPS1组细胞活力、LDH释放率和内皮细胞NT表达差异无统计学意义(P>0.05),LPS10组细胞活力降低,LDH释放率升高,内皮细胞NT表达上调(P<0.01);与LPS10组比较,I40+LPS10组细胞活力、LDH释放率和内皮细胞NT表达差异无统计学意义(P>0.05),P4+LPS10组和P40+LPS10组细胞活力升高,LDH释放率降低,内皮细胞NT表达下调(P<0.01).结论 异丙酚可通过抑制ONOO'-的生成,减轻内毒素诱导人脐静脉内皮细胞损伤.

Effects of propofol on production of peroxynitrite anion in LPS-induced human umbmcal vein endothelial cell injury

Objective To investigate the effects of propofol on the production of peroxynitrite anion (ONOO-) in lipopelysaccharide (LPS)-induced human umbilical vein endothelial cell (HUVEC) injury. Methods HUVECs were provided by centre for preservation of cell strains of special species, Wnhan University, and were assigned to one of 7 groups (n = 5 each). In group Ⅰ no additive was added to HUVECs. In group Ⅱ , Ⅲ ,HUVECs were incubated with LPS 0.1, 1.0, 10 μg/ml. In group Ⅴ, Ⅵ HUVECs were incubated with propofol 4 or40 μg/ml + LPS 10 μg/ml. In group Ⅶ HUVECs were incubated with intralipid (solvent for propofol) 40 μg/ml + LPS 10 μg/ml. After 6 h incubation nitrotyrosine protein in HUVECs was detected by immunocytochemical staining and immunoblot assay. Lactate dehydrogeuase (LDH) release and cell viability were measured. Results LPS significantly increased the expression of nitrotyrosine, enhanced LDH release and decreased live cell numbers in a concentration-dependent manner. These changes were significantly attenuated by propofoi especially 40 μg/ml. Conclusion Propofol can protect HUVECs from LPS-induced injury through inhibiting the production of ONOO-.