生大黄灭菌溶液对脓毒症大鼠的治疗作用及对炎症因子的影响  

Study on applications of rheum sterile solution in rats following sepsis and effect on inflammatory factors

作　　者：巩博[1,2] 江伟伟[1] 魏东坡 何超[1] 王胜云[1] 刘雪峰[1] 李文放[1] 

机构地区：[1]第二军医大学附属长征医院急救科,上海200003 [2]解放军第161医院急诊科,武汉430012

出　　处：《中华急诊医学杂志》2017年第5期544-548,共5页Chinese Journal of Emergency Medicine

基　　金：上海市卫生局中医药科研基金课题（2010Y005A）

摘　　要：目的应用超声波法制备生大黄灭菌溶液，观察其在脓毒症大鼠中的效果，探讨其对于炎症反应的作用机制，为急危重患者救治提供新措施。方法首先以超声法萃取制备生大黄灭菌液。其次将50只雄性SD大鼠随机（随机数字法）分为盲肠结扎穿刺（CLP）组、生大黄组，用盲肠结扎穿刺法模仿中度脓毒症，分别予生理盐水、生大黄灭菌液灌胃，观察7 d生存率。再次将54只大鼠随机（随机数字法）分为3组：假手术组、CLP组、生大黄组，CLP组和生大黄组进行脓毒症造模，术后分别予生理盐水、生大黄灭菌液灌胃。分别在12 h、24 h、48 h每组随机挑选6只大鼠处死，用酶联免疫吸附法（ELISA）检测血清中肿瘤坏死因子-α（tumor necrosis factor-alpha，TNF-α）、高迁移率族蛋白1（high mobility group box-1 protein，HMGB1）水平。用蛋白质印迹法检测小肠组织HMGB1、糖基化终产物受体（receptor for advanced glycation endproducts，RAGE）、核因子-κB（NF-κB） P65蛋白。结果超声萃取法蒽醌成分含量高于于传统水煎法（P〈0.01）。生大黄组大鼠7 d生存率（76%）高于CLP组（48%）（P〈0.05）。生大黄组和CLP组大鼠血清TNF-α、HMGB1水平高于假手术组（P〈0.05）；生大黄组TNF-α水平在各个时间点低于CLP组；在术后12 h生大黄组血清HMBG1水平与CLP组差异无统计学意义（P〉0.05）,在术后24 h、48 h，生大黄组血清HMBG1水平低于CLP组（P〈0.05）。48 h生大黄组和CLP组小肠组织HMGB1、RAGE蛋白水平、NF-κB活化程度高于假手术组（P〈0.01），上述指标CLP组高于生大黄组（P〈0.05）。结论本研究提取出的生大黄灭菌液能下调炎症因子水平，调控炎症反应，提高脓毒症大鼠的生存率。Objective To study the therapeutic effect of rheum （Chinese herbal medicine ） preparation made by using ultrasonic technique on pro-inflammatory cytokines and sepsis in rats. In order to offer novel measure for the treatment of critically ill patients. Methods Firstly, rheum sterile solution was prepared through ultrasonic technique. Secondly, fifty healthy male SD rats were randomly （random number） divided to CLP group and rheum group. Moderate degree of sepsis model was established by using cecal ligation and puncture （CLP）. Rats in group rheum received the liquid rheumpreparation via intragastric administration, while rats in group CLP received saline instead. The 7-day survival rate was recorded and was compared between two groups. In addition, another fifty-four rats were randomly （ random number） divided to sham group, CLP group and rheum group （n = 18 in each group）. CLP was performed to induce sepsis in CLP group and rheum group. Then rats in rheum group received rheum sterile solution via intragastric administration, while rats in CLP group received saline instead. At 12 hours, 24 hours and 48 hours after modeling, six rats in each group were randomly sacrificed. Serum TNF-α and HMGB1 levels were detected by ELISA method. Levels of RAGE, HMGB1 and NF-KB P65 in small intestine were detected by Western Blot. Results Level of anthraquinones extracted from rheum by ultrasonic technique was higher than that by conwentional decoction method. The 7-day survival rate of rats in rheum group （76%） was higher than that in CLP group （48%） （P 〈0. 05）. Compared with sham group, serum TNF-α and HMGB1 levels in CLP group and rheum group were significantly increased （P 〈 0. 05 ）. TNF-α was significantly lower in rheum group than that in CLP group at each interval （P 〈 0. 05） . At 12 hours after modeling, there was no significant difference in serum HMGB1 level between CLP group and rheum group （P 〉 0. 05）. At 24 hours and 48 hours after modeling, serum HMGB1 levels w

关 键 词：超声法 生大黄灭菌溶液 稳定性 脓毒症 促炎因子 

分 类 号：R459.7[医药卫生—急诊医学]