目的 通过检测大鼠小肠平滑肌线粒体凋亡信号通路Bc1-2、Bax以及核转录因子-kB(NF-kB)的蛋白表达,探讨大承气汤促进多器官功能障碍综合征(MODS)胃肠动力恢复的机制。方法 健康成年Wistar大鼠100只,按随机数字表法分为对照组(20只)、模型组(40只)和大承气汤组(40只)。经腹腔注射1mL大肠杆菌混悬液(8×10^8cfu/mL)制备细菌性腹膜炎致MODS大鼠模型,对照组则腹腔注射1mL生理盐水;大承气汤组于制模前2d灌胃大承气汤,每次10mL/kg,每日2次。各组于制模24h后脱颈活杀大鼠取上段小肠,采用免疫组化法检测小肠平滑肌Bcl-2、Bax和NF—KB的蛋白表达。结果 对照组可见小肠平滑肌有大量分布均匀的Bcl-2蛋白表达及少量分布均匀的Bax、NF—kB蛋白表达。与对照组比较,模型组小肠平滑肌Bcl-2蛋白表达分布稀疏,呈散块状区域表达,Bcl-2蛋白表达明显减少(积分A值:7115.3±1797.2比22085.5±4892.2,P〈0.05),以环层肌中尤为明显;Bax、NF—kB蛋白分布密集,表达均明显升高[Bax(积分A值):33802.6±5778.0比7984.4±1804.5,NF—KB(积分A值):2465.9±664.8比1572.6±256.0,均P〈0.05],尤以环行肌中升高明显。与模型组比较,大承气汤组大鼠小肠平滑肌中Bcl-2蛋白表达明显升高(积分A值:12458.6±2491.1比7115.3±1797.2,P〈0.05),Bax、NF—kB蛋白表达明显减少(Bax(积分A值):12529.2±2018.5比33802.6±5778.0,NF—KB(积分A值):1843.1±373.6比2465.9±664.8,均P〈0.05],以环行肌中尤为显著。结论大承气汤可能通过增加线粒体外膜Bcl-2表达,抑制Bax向线粒体膜移位,从而减轻MODS大鼠小肠平滑肌线粒体的损伤,促进胃肠动力的恢复。
Objective To discuss the mechanism of promotion of gastrointestinal motility during multiple organ dysfunction syndrome ( MODS ) by Dachengqi decoction, by examining the expression of Bcl-2, Bax of mitochondrial pathway, and nuclear factor-kB (NF-kB) in smooth muscle of the small intestinal in rats. Methods According to the random number table, 100 healthy adult Wistar rats were divided into three groups: control group with 20 rats, model group with 40 rats, and Dachengqi decoction group with 40 rats. Rat model of MODS was reproduced by bacterial peritonitis induced by an injection of 1 mL Escherichia coli suspension (8×10^8 cfu/mL) into peritoneal cavity. The rats in control group were given 1 mL normal saline intraperitoneally. The rats in Dachengqi decoction group were given 10 mL/kg Dachengqi decoction by garage, twice a day, before inoculation of the bacterial suspension. Twenty-four hours after modelling, rats in all groups were sacrificed by cervical vertebra luxation, and the upper small intestine was harvested to detect the protein expressions of Bcl-2, Bax, and NF-KB in smooth muscle tissue using immunohistochemical staining. Results In the control group, a large amount of Bcl-2 protein was expressed and it was distributed uniformly in small intestinal smooth muscle. On the other hand, a small amount of Bax and NF-KB protein was expressed, and they were also distributed uniformly. Compared with the control group, Bcl-2 protein was distributed only sparsely, and it was scattered in intestinal smooth muscle in blocks in the model group. The expression of Bcl-2 protein was obviously down-regulated [ integral optical density (A) value: 7 115.3 ± 1 797.2 vs. 22 085.5±4 892.2, P 〈 0.05], and this pheiiomenon was more prominent in circular muscle layer. Bax and NF-kB were densely distributed, and their expressions were upgraded obviously [Bax (A value): 33 802.6± 778.0 vs. 7 984.4±1 804.5, NF-kB(A value): 2 465.9±664.8 vs. 1 572.6±256.0, both P 〈 0.05 ]. This phenomen