凉膈散抑制创伤失血性休克大鼠肝脏实质细胞凋亡的机制研究  

The mechanism of Liangge San （凉膈散） for inhibiting apoptosis of hepatic parenchymai cells in rats with traumatic hemorrhagic shock ZHAO Zhi-ling, CAO Shu-hua, WANG Yong-qiang. The First Central

作　　者：赵志伶[1] 曹书华[1] 王勇强[1] 

机构地区：[1]天津医科大学第一中心临床学院,天津市第一中心医院急救医学研究所,天津300192

出　　处：《中国中西医结合急救杂志》2011年第2期82-85,I0001,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：天津市医药卫生科研基金资助项目（05KZE17）

摘　　要：目的观察凉膈散对创伤失血性休克大鼠肝脏细胞凋亡的影响，并探讨其机制。方法健康雄性Wistar大鼠90只，随机分为正常组，模型1、3、6、24h组和凉膈散1、3、6、24h组，每组10只。采用骨折后经颈动脉放血制备创伤失血性休克大鼠模型。凉膈散组于制模前连续3d灌胃凉膈散10ml／kg；模型组给予等量生理盐水；正常组不予任何处理。取血，采用速率法测定血浆天冬氨酸转氨酶（AST）浓度；取肝组织，用原位末端缺刻标记法（TUNEL）检测凋亡细胞，用免疫组化法检测天冬氨酸特异性半胱氨酸蛋白酶3（caspase-3）蛋白表达，用逆转录-聚合酶链反应（RT-PCR）检测Bax、Bcl一2、Fas、FasL的基因表达。结果模型组1、3、6、24h血AST（U／L）呈上升趋势，24h达峰值（647．36±60．02），与正常组（66．69±19．95）比较差异有统计学意义（均P〈0．05）；凉膈散组各时间点血AST（194．24±29．53、306．01±83．85、388．36±92．71、451．48±99．70）均较模型组显著降低（均P〈0．05）。正常组未见凋亡细胞；模型组1、6、24h时有散在凋亡细胞，3h时中央静脉周围肝脏实质细胞出现大量凋亡（个：20．60±0．47）；凉膈散组3h时只发现肝窦内淋巴细胞凋亡（个：14．70±1．02），较模型组显著降低（P〈0．05）。模型组1、3、6和24hcaspase-3蛋白表达（×10^3，A值）均较正常组（8．96±1．22）显著升高，3h达峰值（49．71±0．89）；凉膈散组各时间点caspase-3表达（17．81±0．48、34．48±1.09、28.52±1．14、31．76±1．39）较模型组显著下降（均P〈O．05）。模型组各时间点凋亡相关基因表达均较正常组升高，且Bax、Bcl-2、Fas均于3h达峰值（1．98±0．07、0．87±0．07、0．25±0．02），FasL于6h达峰值（0．57±0．02）；凉膈散组在3h（0．40±0．03）和6h（0．18±0．03）能显著降低Bax基因表达（均P〈0．05），对BcObjective To observe the effect of Liangge San （凉膈散） on apoptosis of liver cells in rats with traumatic hemorrhagic shock and to discuss the mechanism. Methods Ninety healthy male Wistar rats were divided randomly into nine groups ： normal group, model 1-, 3-, 6-, 24-hour groups and Liangge San 1-, 3-, 6-, 24-hour groups （each, n=10）. By bloodletting from carotid artery after bone fracture, the rat model of traumatic hemorrhagic shock was established. The rats in Liangge San groups were infused into the stomach with Liangge San （10 ml/kg） once a day for 3 days before the model establishment, the rats in model groups were infused with equal amount of 0. 9% normal saline （NS） and no treatment was given in normal group. The plasma was collected to measure aspartate aminotransferase （AST） concentrations with Rate method. The liver was harvested to detect the cell apoptosis with the terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling （TUNEL） assay, the expression of caspase-3 protein by the techniques of immunohistochemistry, and the mRNA expressions of Bax, Bcl-2, Fas, FasL with reverse transcriptase- polymerase chain reaction （RT-PCR）. Results AST （U/L） in model 1-, 3-, 6-, 24-hour groups was increased gradually, and peaked at 24 hours （647.36±60.02） with statistically significant difference compared with that of the normal group （66.69±19.95, all P〈0.05）. The levels of AST in Liangge San groups at all time points （194.24 ± 29.53, 306.01 ± 83.85, 388.36 ± 92.71, 451.48± 99.70） were obviously lower than those in the model groups （all P〈0.05）. No apoptosis was found in normal group; in model 1-, 6-, 24-hour groups there were scattered apoptotic cells, and in the 3-hour model group, the apoptosis in a large amount was around the central vein （20.60± 0.47）; the count of apoptotic lymphocytes in the sinusoid in Liangge San 3-hour group （14.70±1.02） was less significantly compared with the model 3-hour group （P〈0. 05）. The

关 键 词：凉膈散 休克 创伤失血性 肝脏 凋亡 

分 类 号：R285.5[医药卫生—中药学] R541.64[医药卫生—中医学]