脂多糖致脓毒血症大鼠血管内皮细胞凋亡的实验观察

Observation of Vascular Endothelium Cell Apoptosis in Endotoximia Rat induced by LPS

目的:观察细菌脂多糖(LPS)致脓毒血症大鼠血管内皮细胞(VEC)凋亡的器官靶向性及时相性特征。方法:70只SD大鼠经尾静脉一次性注射LPS(10mg/kg)复制脓毒血症模型后平分为30min组、1h组、2h组、4h组、8h组、16h组和24h组。另选10只SD大鼠作为正常对照组。眼眶采血,ELISA检测各组血清中血管性血友病因子(vWF)及炎性介质白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的变化特征。断头处死对照组、1h组、4h组和8h组大鼠,取肺、肝、心、脑、肾制作组织切片,采用地高辛标记凋亡细胞,免疫组化染色检测上述各组大鼠各脏器VEC凋亡。结果:2h-24h组TNF-α浓度均较对照组升高,4h组达峰值(P<0.01);IL-1β在2h组和4h组显著高于对照组,2h组为峰值(P<0.01),8h-24h组与对照组无显著差异(P>0.05)。4h-8h组大鼠血清vWF浓度较对照组升高,4h组达峰值(P<0.05),其它时相组与对照组差异无统计学意义(P>0.05)。LPS致脓毒血症大鼠肺、肝、心、脑、肾存在不同程度VEC凋亡现象,其中肺部受累较早。结论:LPS(10mg/ml)所致脓毒血症大鼠重要脏器可出现VEC凋亡,以肺部较早。

Objective：To evaluate vascular endothelium cell （VEC）apoptosis and its characteristics in lipopo-lysaccharide （LPS）induced endotoxic model in rat.Method：LPS （10mg/kg）was administered in one bout via cau-dal vein in 70 rats and then divided randomly into LPS-administered after 30min,1h,2h,4h,8h,1 6h,and 24h group.Another 10 rats was adopted as control group.Blood was collected from orbit.The concentrations of von Willebrand factor （vWF）and interleukin 1β（IL-1β）and tumor necrosis factorα（TNF-α）were detected by ELISA. Rats were sacrificed by decaptitation.The tissue of heart,liver,brain,kidney,lung were collected and made tissue slice.VEC apoptosis of these organs was evaluated with immunohistochemisty staining labeled with digoxin dUTP after LPS was administered.Results：The concentration of TNF-αin serum increased in LPS-administered 2h-24h group compared with the control group,and peaked at 4h group （P 〈0.01）.The concentration of IL-1βin serum al-so increased in LPS- administered 2h- 4h group,peaked at 2h group （P 〈 0.01 ）,and there were no significant difference between 8h-24h and control group respectivly （P 〉0.05）.The concentration of vWF in serum enhanced gradually after LPS in LPS-administered 2h-8h group,peaked at 4h group （P 〈0.05）,decreased in 1 6h and 24h group compared with 4h group （P 〈0.05）.The apoptosis of VEC from vital organ （lung,heart,brain,kidney and liver）occurred after LPS administered for 2-4h,and lung was the most early organ involved.Conclusion：VEC ap-optosis occurred in LPS （10mg/ml）induced endotoximia in rat,and lung is the much earlier organ involved.