灯盏花素对脑死亡巴马小型猪肾脏的保护作用  

Effects of breviscapine on the renal structure, function and PKC-mRNA and its protein expression in brain-dead BA-Ma mini pigs

作　　者：史冀华[1] 王海溥[1] 唐哲[1] 赵永福[1] 张水军[1] 

机构地区：[1]郑州大学第一附属医院器官移植中心,450052

出　　处：《中华器官移植杂志》2006年第12期755-758,共4页Chinese Journal of Organ Transplantation

基　　金：河南省杰出人才创新基金资助项目(0421002500)

摘　　要：目的探讨灯盏花素对脑死亡状态下巴马小型猪的肾脏结构、功能与蛋白激酶C (PKC)-αmRNA及其蛋白表达的影响,并研究其可能机制。方法将15只巴马小型猪随机分为3组,每组5只。A组:应用改进的缓慢间断颅内加压法建立脑死亡模型,不行药物干预;B组:采用与A组相同的方法建立脑死亡模型,分别于初次确认脑死亡后1和12 h经静脉缓慢滴注灯盏花素2．5 mg/kg;C组:不建立脑死亡模型,动物麻醉后仅行开颅与开、关腹手术,检测各项相关数据作为对照。分别于首次判定脑死亡后3、6、12、18和24 h检测巴马小型猪血清尿素氮(BUN)和血肌酐(Cr)及炎性因子IL-1β、IL-6、TNT-α的表达水平。于脑死亡后3、6、12及24 h开腹取相同部位肾脏组织,光镜下观察肾脏组织结构变化,电镜下观察肾脏超微结构变化,免疫组织化学染色观察PKC-α的表达水平,逆转录聚合酶链反应(RT-PCR)检测PKC-αmRNA的动态变化。结果(1)A组及B组血清IL-1β、IL-6、TNF-α水平自脑死亡后3 h开始逐渐升高,血清BUN和Cr水平自脑死亡后12 h开始逐渐升高。每一时点与前一时点比较,差异均有统计学意义(P<0．05);但B组升高幅度显著低于A组(P<0．05)。(2)A组及B组肾组织中PKC-αmRNA及其蛋白表达水平自脑死亡后3 h开始逐渐升高,每一时点与前一时点比较,差异均有统计学意义(P<0．05)。(3)脑死亡后12 h可见肾组织细胞结构变化;B组肾组织细胞结构变化明显轻于A组。结论灯盏花素能够抑制肾组织中PKC-αmRNA及其蛋白的表达,减少炎症介质的释放,从而保护脑死亡状态下肾脏的功能及结构。Objective To investigate the effects of breviscapine on the renal structure, function and PKC-α mRNA and its protein expression in brain-dead BA-Ma mini pigs. Methods Fifteen BA-Ma mini pigs were randomly divided into 3 groups： brain-dead group （group A）, breviscapine pretreatment group （group B）, and control group （group C）, 5 pigs in each group. The brain-dead models were established by increasing intracranial pressure in a modified, slow and intermittent way. At 3, 6, 12, 18 and 24 h after the initial brain death, serum BUN, Cr, TNF-α, IL-1β, and IL-6 were determined. At 3, 6, 12, and 24 h, the changes of renal tissues were observed by HE staining, and the expression of PKC-α mRNA and protein was detected by RT-PCR and immnohistochemistry respectively. The ultrastructural changes of hepatic ceils were observed under electron microscopy. Results （1） At 3rd h after the initial brain death, IL-1β, IL-6, and TNF-α in group A and group B began to increase. Serum BUN and Cr in group A and group B began to in crease at 12 th after brain death and were higher at each time point （P〈0. 05）. Inflammatory factors at 3, 6, 12, 18, and 24 h had significant differences （P〈0.05）. BUN and Cr in group B after 12 h were significantly different from those in group A （P〈0. 05）. In group A, these parameters were significantly higher than in group B at each time point （P〈0. 05）. （2） The expression of PKC-α mRNA and protein in groups A and B began to increase at 3rdh. The expression of PKC-α mRNA and protein at 6, 12, and 24 h showed significant differences （P〈0. 05）, which was significantly higher in group A than in group B at each time point （P〈0.05）. （3） After 12 h, changes of renal cells could be found, the injury degree of hepatic cells in group B were milder than that in group A. Conclusion Breviscapine can inhibit the expression of PKC a mRNA and protein, decrease the release of inflammatory factors, and then alleviate the renal injury during brain death.

关 键 词：灯盏细辛 脑死亡 肾脏 猪 

分 类 号：R285.5[医药卫生—中药学]