z-VAD-FMK与Necrostatin-1联合对MODS大鼠器官的保护作用研究

Protective effect of z-VAD-FMK and Necrostatin-1 on the organs of rats with MODS

目的:探讨z-VAD-FMK与Necrostatin-1联合对MODS大鼠器官的保护作用。方法:采用失血性休克-内毒素"二次打击"建立大鼠MODS动物模型。将80只雄性SD大鼠随机分为4组:z-VAD-FMK组、Nec-1组、z-VAD-FMK+Nec-1组、模型组,每组20只,观察72 h死亡率。另取40只雄性SD大鼠随机分为5组:假手术组、z-VAD-FMK组、Nec-1组、z-VAD-FMK+Nec-1组、模型组,每组8只,分别于注射内毒素和生理盐水后24 h收集血清和肝、肺、肠组织。全自动生化仪检测血清ALT、AST的变化,血气分析仪检测PaO2的变化,ELISA检测血清TNF-α、IL-1β的变化,HE染色观察肝、肺、肠组织的病理学改变。结果:大鼠遭受失血性休克-内毒素"二次打击"后,血清ALT、AST明显升高(P<0.01),PaO2显著下降(P<0.01),血清炎性因子TNF-α、IL-1β水平明显升高(P<0.01)。光镜下见肝窦明显扩张、淤血,肝小叶结构紊乱,肝细胞部分变性、坏死;肺泡壁结构被破坏,肺间质充血,大量炎性细胞浸润;肠绒毛结构被破坏,部分绒毛顶部细胞、隐窝细胞变性、坏死,大量炎性细胞浸润。而z-VADFMK组、Nec-1组及z-VAD-FMK+Nec-1组ALT、AST升高程度、PaO2下降程度、TNF-α、IL-1β升高程度均显著低于模型组(P<0.01),以z-VAD-FMK+Nec-1组变化最明显。72 h死亡率均较模型组显著降低(P<0.05)。结论:z-VAD-FMK、Nec-1、z-VADFMK+Nec-1联合治疗均可显著减轻MODS大鼠的器官损伤,联合治疗效果更佳。

Objective： To investigate the protective effect of z-VAD-FMK and Necrostatin-1 on the organs of rats with MODS. Methods： Aduh male SD rats were used and rat model was produced by hemorrhagic shock based on injecting LPS. A total of 80 rats were randomly divided into four groups： z-VAD-FMK group; Nec-1 group; z-VAD-FMK＋Nec-1 group and model group with 20 rats in each group. And the 72 h mortality was observed. In addition, other 40 rats were randomly divided into five groups： sham group; z-VAD-FMK group; Nec-1 group; z-VAD-FMK＋Nec-1 group; model group, with 8 rats in each group. The samples of different groups were collected at 24 h after injecting LPS or normal saline. Serum ALT and AST were detected by automatic biochemistry analyzer. PaO2 was detected by blood gas analyzer. And serum TNF-ct and IL-1β were detected by ELISA. The pathological changes of liver, lung and intestine were observed by H＆E staining. Results： After hemorrhagic shock, the expression of ALT, AST, TNF-a and IL-1β significantly increased （P〈0.01） while the level of PaO2 markedly reduced （P〈0.01）. Under light microscopy, hepatic sinus expansion, liver ceils degeneration, necrosis, as well as infiltration of abundant inflammatory cells were observed; the structure of alveolar walls were destroyed and the pulmonary interstitial hyperemia and infiltration of abundant inflammatory cells were observed; the structure of intestinal villi were destroyed and the top part of villi cells and crypt cells degeneration, necrosis, as well as infiltration of abundant inflammatory cells were observed. Compared with model group, the up-regulation degree of ALT, AST, TNF-α and IL-1β and the PaO2 reduction were smaller in z-VAD-FMK group, Nec-1 group and z-VAD-FMK＋Nec-1 group （P〈0.01）. And the changes in z-VAD-FMK＋Nec-1 group were the most significant. In addition, the 72 h mortality significantly reduced （P〈0.05）. Conclusion： The z-VAD-FMK, Nec-1 and z-VAD-FMK＋Nec-1 can significantly reduce the organ damage of rats with MODS, and combining z-VAD-FMK and Nec-1 can bring marked results.