通腑泻肺方对脓毒症急性肺损伤大鼠NLRP3调控作用的实验研究  

Experimental Study of Tongfuxiefei Decoction on Regulating NLRP3 in Rats with Sepsis in Acute Lung Injury

作　　者：张涛[1] 熊旭东[1] 赵敏[1] 

机构地区：[1]上海中医药大学附属曙光医院,上海200021

出　　处：《中国中医急症》2014年第8期1406-1408,1440,共4页Journal of Emergency in Traditional Chinese Medicine

基　　金：高等学校博士学科点专项科研基金(20113107110003)

摘　　要：目的观察通腑泻肺方对脓毒症急性肺损伤大鼠NLRP3炎性体表达的影响,探索通腑泻肺方治疗脓毒症急性肺损伤的作用机制。方法采用盲肠结扎穿孔法(CLP)复制脓毒症急性肺损伤动物模型。将32只清洁级健康成年雄性SD大鼠随机分为空白对照组、假手术组、模型组和治疗组,每组8只。空白对照组麻醉后腹主动脉采血致死;假手术组麻醉后开腹,翻动胃肠后关腹,24 h后麻醉并腹主动脉采血致死;模型组于CLP后立即予生理盐水2 mL灌胃1次,CLP后6 h、12 h,再分别给予生理盐水2 mL灌胃1次,CLP后24 h予麻醉并腹主动脉采血致死;治疗组于CLP后立即予通腑泻肺方中药灌胃1次,CLP后6 h、12 h,再分别给予通腑泻肺方中药灌胃1次,CLP后24 h予麻醉并腹主动脉采血致死。观察各组大鼠的一般表现,ELISA法检测各组血清中Caspase-1含量,Western blot法检测各组大鼠肺组织中NLRP3、ASC含量,RT-PCR法检测肺组织中NLRP3、ASC的mRNA表达量。结果模型组和治疗组肺组织NLRP3、ASC的蛋白水平、mRNA表达水平以及血清中caspase-1水平均显著高于空白对照组和假手术组(P<0.01);模型组肺组织NLRP3、ASC的蛋白水平、mRNA表达水平以及血清中Caspase-1水平显著高于治疗组(P<0.01)。结论通腑泻肺方能够下调脓毒症急性肺损伤大鼠NLRP3、ASC、Caspase-1的表达水平,具有调控NLRP3炎性体合成的作用。Objectives： To observe the effect of Tongfuxiefei decoction on expression of inflammasome NLRP3 in rats with sepsis in acute lung injury（ALI） and to explore the mechanism of Tongfuxiefei decoction in sepsis treatment. Methods： The animal model of sepsis in acute lung injury was made by cecal ligation and puncture（CLP）. 32 male SD rats were randomly divided into four groups： control group,sham operation group,model group and treatment group. Each group has 8 rats. The control group rats were sacrificed by extracting the blood from abdominal aorta after anesthesia. The sham operation group rats were opened the abdomen and turned over intestinal tract after anesthesia,then closed the abdomen,got the blood 24 h later after anesthesia. The model group rats were given 2ml normal saline and the treatment group rats were given Tongfuxiefei decoction by intragastric ad- ministration 6 h and 12 h after CLP. Rats in both groups were sacrificed by extracting the blood from abdominal aorta at 24h later after CLP. Observe the general performance of each group. Enzyme-linked immunosorbent assay was used to detect the level of Caspase-1 in serum. The protein and the mRNA expressions of NLRP3 and ASC in lung tissue were respectively detected by Western blot method and reverse transcription-polymerase chain reaction （RT-PCR）. Results： The protein levels and mRNA expressions of NLRP3 and ASC,and Caspase-1 levels of the model group and of the treatment group were significantly higher than those of the control group and of the sham operation group （P〈 0.01）. The protein levels and mRNA expressions of NLRP3 and ASC,and Caspase-1 levels of model group were significantly higher than those of treatment group （P〈 0.01 ）. Conclusions： Tongfuxiefei decoction can reduce the expression of NLRP3 and ASC as well as the level of Caspase-1 ,so as to regulate NLRP3 synthesis.

关 键 词：脓毒症 通腑泻肺方 炎性体 急性肺损伤 

分 类 号：R285.5[医药卫生—中药学]