重楼总皂苷对脓毒症大鼠的保护作用研究

Protective effects of rhizoma paridis total saponins （重楼总皂苷） on septic rats

目的观察重楼总皂苷对脓毒症大鼠的保护作用并探讨其作用机制。方法采用盲肠结扎穿孔术（CLP）制备大鼠脓毒症模型，观察重楼总皂苷对CLP大鼠死亡率的影响。用酶联免疫吸附法（ELISA）检测术后2、6、12、24和48h大鼠血清肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）水平；光镜下观察72h大鼠肺组织病理学变化。分离大鼠腹腔巨噬细胞（PMФ），用ELISA检测重楼总皂苷对LPS（100μg／L）诱导大鼠PM由释放TNF-α及IL-1β的影响。结果重楼总皂苷组大鼠死亡率明显低于模型组（50．0％比85．0％，P〈0．05）。重楼总皂苷各浓度组血清TNF-α和IL-1β水平较模型组相应时间点明显降低（P〈0．05或P〈0．01）；肺组织病理学观察显示肺损伤程度较轻。体外实验表明，5、10、20和40mg／L重楼总皂苷均可显著降低LPS刺激PMФ释放TNF-α和IL-1β的水平（P均〈0．01）；且各浓度组间TNF-α水平差异无统计学意义（P均〉0．05）；而5mg／L组IL-1β水平显著高于10mg／L组（P〈0.05），但与20mg／L组和40mg／L组比较差异无统计学意义（P均〉0．05）。结论重楼总皂苷能抑制CLP大鼠巨噬细胞活化并减轻肺损伤程度，对CLP大鼠具有保护作用。

Objective To investigate the protective effect of rhizoma paridis total saponins （重楼总皂苷） and its mechanism on septic rats. Methods Septic model was reproduced by cecal ligation and puncture （CLP） in Wistar rats. Rhizoma paridis total saponins was administered to observe its protective effects on septic rats. Blood was collected to determine serum tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） levels at 2, 6, 12, 24 and 48 hours after operation by means of enzyme-linked immunosorbent assay （ELISA）. The pathological changes of lung tissue were observed with light microscope at 72 hours after operation. The peritoneal macrophages （PMФ） in rats were isolated and the release of TNF-α and IL-1β in PMФ after exposure to lipopolysaccharide （LPS, 100 μg/L） were measured by ELISA. Results Mortality in the rhizoma paridis total saponins group was significantly lower than the CLP group （50.0% vs. 85.0%, P〈0. 05）. The levels of TNF-α and IL-1β in serum were significantly lower than those of the CLP group at the same time （P〈0.05 or P〈0.01）. The degree of inflammatory injury to the lung was much milder than that in the CLP group. In the in vitro experiment, it was shown that rhizoma paridis total saponins in concentrations of 5, 10, 20 and 40 mg/L could inhibit remarkably the release of TNF-α and IL-1β from LPS-stimulated PMФ of rats （all P〈0.01）. The differences in TNF-α levels among the groups showed no statistically significant difference （all P〉0.05）. The level of IL-1β in 5 mg/L group was significantly higher than that of the 10 mg/L group （P〈0. 05）, but showed no difference with those of 20 mg/L and 40 mg/L groups （both P〉0.05）. Conclusion Rhizoma paridis total saponins can protect the CLP rats by inhibiting the activation of rat PMФ to release cytokines and ameliorating acute lung injury.