乌司他丁对脓毒症大鼠血管内皮细胞损伤保护作用及机制研究  

Mechanism and protective effect of ulinastatin on vascular endothelial cells in sepsis rats

作　　者：樊楚明[1] 杨欣悦[1] 任靖宇[1] 张洪波[1] 茹今 陈庆宁[2] 

机构地区：[1]云南省第一人民医院重症医学科,昆明650032 [2]云南省第一人民医院皮肤科,昆明650032

出　　处：《中国临床药理学杂志》2016年第8期723-726,共4页The Chinese Journal of Clinical Pharmacology

基　　金：云南省科技厅昆明医科大学联合基金资助项目(2014FZ066)

摘　　要：目的研究乌司他丁对脓毒症大鼠血管内皮细胞(VEC)损伤保护作用及机制研究。方法 40只SD雄性大鼠按体重随机分为4组:假手术组,模型组及2个剂量实验组(乌司他丁,10×10~4,20×10~4U·kg^(-1)),其中模型组及实验组应用盲肠结扎穿孔(CLP)构建脓毒症大鼠模型。免疫酶联吸附法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及IL-6含量;试剂盒法检测血清一氧化氮(NO)及诱导型一氧化氮合酶(iNOS)含量;流式细胞术检测VEC凋亡情况;分光光度法检测血清中含半胱氨酸的天冬氨酸水解酶3(Caspase 3)活性;免疫印迹法分析各组B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)含量变化及核因子-κB(NF-κB)信号通路的激活状况。结果与假手术组比较,模型组中TNF-α、I L-1β、IL-6、NO、iNOS含量与VEC凋亡率及Caspase 3的活性均显著提高,Bax表达量明显上升,而Bcl-2表达量明显下降,NF-κB p65及IκBα磷酸化水平明显提高(均P<0.01);与模型组比较,2个剂量实验组皆能显著抑制上述指标的变化(P<0.05)。其中Caspase 3活性比值为(1.00±0.21,3.70±0.25,1.83±0.14,1.53±0.13)。结论乌司他丁对脓毒症大鼠VEC损伤具有保护作用,这可能与NF-κB信号通路有关。Objective To explore the mechanism and protective effect of ulinastatin on vascular endothelial cells（VEC） in sepsis rats. Methods Forty SD male rats were randomly divided into four groups： sham operation group,model group and low and high dose test groups（ulinastatin,10 × 10^4,20 × 10^4 U · kg^-1）. The sepsis rat models were established with cecal ligation and puncture（CLP）. The level of tumor nuclear factor-α（TNF-α）,interleukin 1β（IL-1β） and IL-6 in serum was assayed by enzyme linked immunosorbent assay method. The level of nitrogen（NO） and inducible nitric oxide synthase（i NOS） was determinated by kits. VEC apoptosis was detected by flow cytometry. The activity of cysteinyl aspartate specific proteinase 3（Caspase 3） was measured by spectrophotometry. The expression of B-cell lymphoma-2（Bcl-2）,Bcl-2 associated X protein（Bax） and the activation of nuclear factor kappa B（NF-κB） signal pathway were assayed by Western blot.Results Compared with sham operation group,the level of TNF-α,IL-1β,IL-6,NO,i NOS was higher,the number of apoptotic VECwas higher,the activity of Caspase 3 and the expression of Bax was higher,the expression of Bcl-2 was lower,the phosphorylation of NF-κB p65 and IκBα was higher in the model group,the difference was significant（all P〈0. 01）. Compared with model group,low and high dose test groups could inhibit the change,the difference was significant（P〈0. 05）. The ratio of Caspase 3 activity was（1. 00 ± 0. 21,3. 70 ± 0. 25,1. 83 ± 0. 14,1. 53 ± 0. 13）.Conclusion These results suggested ulinastatin inhibited the injury of VEC in sepsis rats via resisting to inflammation and VEC apoptosis,which might be associated with NF-κB signal pathway.

关 键 词：乌司他丁 脓毒症 血管内皮细胞 炎症 凋亡 

分 类 号：R976[医药卫生—药品]