大黄附子汤对大鼠失血性休克复苏后肠黏膜屏障的保护作用

Protective effect of Dahuangfuzi decoction on intestinal barrier after hemorrhagic shock in rats

目的探讨大黄附子汤对大鼠失血性休克(hemorrhagic shock,HS)复苏后小肠黏膜屏障功能保护作用的机理。方法健康清洁级雄性SD大鼠,72只,随机数字表法分为对照组、模型组、治疗(大黄附子汤)组,各组再按HS模型制备成功复苏后0、6、12 h分为3个亚组,每组8只。模型组、治疗组采用Wiggers改良法制备HS大鼠模型,模型组以0.9%等渗盐水保留灌肠,治疗组予以大黄附子汤保留灌肠。分别于上述三个时段测定血清中肠脂肪酸蛋白(IFABP)水平及内毒素(LPS)含量;光镜观察大鼠肠黏膜病理改变;Western blot与免疫组化法测定黏膜闭锁小带蛋白-1(zonula occludens-1,ZO-1)、磷酸化血管扩张刺激磷蛋白(phospho-VASP,p-VASP)蛋白表达。结果治疗组与模型组比较,随时间推移肠道血流增加[(119.28±6.50)bpu vs(99.99±0.81)bpu,P<0.01],血清内毒素(LPS)、脂肪酸结合蛋白(IFABP)含量逐渐减低[(21.23±3.33)ng/m L vs(27.09±3.55)ng/m L,P<0.01;(0.420±0.059)ng/m L vs(0.645±0.058)ng/m L,P<0.01],肠道黏膜损伤程度逐渐减轻[(1.78±0.24)vs(3.64±0.25),P<0.01],p-VASP蛋白表达下调[(0.106±0.030)vs(0.297±0.045),P<0.01],ZO-1表达上调[(0.82±0.047)ng/m L vs(0.61±0.041)ng/m L,P<0.01]。结论大黄附子汤可减轻失血性休克复苏后肠道粘膜屏障损伤,可能与调控p-VASP、ZO-1蛋白表达有关。

Objective To investigate the protective effect of Dahuangfuzi decoction on intestinal epithelial tight junction after hemorrhagic shock.Methods Seventy-two SD rats were allocated to sham operation group,hemorrhagic shock controlgroup,Dahuang Fuzi Decotion group according to random number table,and each group was divided into three subgroups according to the time point of 0,6,1 2 h after HS and resuscitation.The rat model of hemorrhagic shock was induced by using the improved Wiggers method. Model group and Dahuang Fuzi Decotion group were given physiological saline or Dahuang Fuzi Decotion 2ml retention enema after the post-resuscitation.At three time points,the level of intestinal fatty acid binding protein (IFABP)in the serum was tested by ELISA method,and the levels of serum endotoxin (LPS)was detected by dynamic turbidimetric method.The morphological changes of small intestinal mucosa in rats was observed under light microscope,while p-VASP and ZO-1 expression was observed by Western blot and immunohistochemical method.Results Compared with model group,the intestinal blood flow was increased [(1 1 9.28 ±6.50)bpu:(99.99 ±0.81 )bpu,P<0.01 ],serum endotoxin (LPS),fatty acid binding protein (IFABP)content gradually reduced [(21 .23 ±3.33)ng/mL:(27.09 ±3.55)ng/mL,P<0.01;(0.420 ±&nbsp;0.059)ng/mL:(0.645 ±0.058)ng/mL,P<0.01 ]were reduced,and the intestinal mucosa damage[(1 .78 ±0.24):(3.64 ±0.25),P<0.01 ]was alleviated,p-VASP protein expression [(0.1 06 ±0.030):(0.297 ± 0.045 ),P<0.01 ],ZO-1 expression [(0.82 ±0.047 )ng/mL:(0.61 ±0.041 )ng/mL,P<0.01 ]was up-regulated.Conclusion Dahuangfuzi decoction can significantly protect the intestinal mucosal barrier function by regulating p-VASP,ZO 1 protein expression after hemorrhagic shock and resuscitation.