目的探讨大承气颗粒对多器官功能障碍综合征(multiple organ dysfunction syndrome,MODS)大鼠小肠平滑肌细胞(smooth muscle cell,SMC)凋亡的干预作用及其机制。方法健康成年Wistar大鼠100只,随机分为对照组(20只)、模型组(40只)和大承气颗粒组(中药组,40只)。模型组和中药组大鼠予腹腔注射大肠希氏菌(E.coli)混悬液,建立细菌性腹膜炎致MODS模型,中药组造模前2天给予大承气颗粒(1mL/100g)灌胃,2次/天,连续3天。造模24h后取存活大鼠的上段小肠组织,分别进行末端脱氧核苷酸转移酶介导的dUTP缺口标记技术(terminal-deoxynucleotidyl transferase mediated nick end labeling,TUNEL)和免疫组织化学染色,观察大承气颗粒干预前后MODS大鼠小肠SMC的凋亡数量及线粒体凋亡信号途径B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2associated X protein,Bax)、细胞色素C(cytochrome c,Cyt c)蛋白表达的变化。结果与对照组比较,模型组小肠SMC凋亡数量、Bax、Cyt c蛋白表达明显升高(P<0.01),Bcl-2蛋白表达明显减少(P<0.01)。与模型组比较,中药组小肠SMC凋亡数量、Bax、Cyt c蛋白表达明显降低(P<0.01),Bcl-2蛋白表达明显升高(P<0.01)。结论大承气颗粒可以通过抑制小肠SMC线粒体凋亡途径的激活,抑制SMC的凋亡,从而促进MODS大鼠胃肠运动功能的恢复。
Objective： To observe the morphological changes in enteric nerve system （ENS） of rats with multiple organ dysfunction syndrome （MODS） treated by Dachengqi Decoction （大承气汤, DCQD）. Methods： Fifty Wistar rats were randomly assigned to the control group, MODS model group and DCQD treated group. The rats in MODS model group and DCQD treated group were injected Escherichia coil （E. coli） suspension into abdominal cavity under sterile condition. The DCQD treated group was gavaged with DCQD 2 days before the E. coil suspension was injected. Twenty-four hours after injection, the proximal segment of intestine was resected and studied by immunohistofluorescence using vesicular acetylcholine transporter, vasoactive intestinal polypeptide （VIP）, substance P （SP） and neuronal nitric oxide synthase （nNOS） antibodies. The whole-mount preparations were observed by laser scanning confocal microscope to detect the changes of quantity and fluorescence integral optical density （IOD） value of intestine enteric nerves. Results： Compared with the control group, the quantity and IOD value of acetylcholine （ACh）, VIP, SP and nitric oxide （NO） nerves of intestine in the MODS group were significantly decreased （P〈0.01）, and the network of enteric nerves was remarkably disrupted. Compared with the MODS group, the quantity and fluorescence IOD value of ACh, VIP, SP and NO nerves in the DCQD group were significantly increased （P〈0.01）, and the network of enteric nerves was remarkably recovered. Conclusions： DCQD can protect and repair damage in the network of ACh, SP, NO and VIP nerves in rats with MODS, which may be one of mechanisms involved in promoting gastrointestinal motility by DCQD.
