摘要
【目的】观察和比较调胃承气汤灌胃与灌肠对肠源性脓毒症(GOS)大鼠肠黏膜屏障的影响。【方法】将48只大鼠随机分为假手术组,模型组,中药灌胃高、低剂量组(分别给予调胃承气汤9.450、4.725 g/kg灌胃),中药灌肠高、低剂量组(分别给予调胃承气汤9.450、4.725 g/kg灌肠),每组8只。采用盲肠结扎穿刺法(CLP)复制GOS大鼠模型。各组在造模后2、10、24 h分别对应给药。采用酶联免疫吸附分析(ELISA)检测血清降钙素原(PCT)、D-乳酸(D-LA)水平,逆转录实时定量PCR(RT-qPCR)法检测小肠组织occludin、闭锁小带蛋白(ZO-1)基因及肝脏组织肿瘤坏死因子α(TNF-α)、干扰素γ(INF-γ)基因表达水平,免疫组化法检测小肠组织occludin、ZO-1蛋白的表达,苏木素—伊红(HE)染色观察小肠病理组织结构的变化。【结果】与假手术组比较,模型组大鼠小肠组织结构有明显损伤,血清PCT和D-LA水平显著升高(P <0.01),小肠组织occludin、ZO-1基因相对表达水平明显下调(P <0.01),肝脏组织TNF-α、INF-γ基因相对表达水平显著上调(P <0.01),小肠组织occludin、ZO-1蛋白免疫组化阳性表达明显减少(P <0.05);与模型组比较,中药各治疗组能明显改善小肠的损伤程度,显著降低血清PCT和D-LA水平(P <0.05或P <0.01),上调小肠组织occludin和ZO-1基因相对表达水平,下调肝脏组织TNF-α和INF-γ基因相对表达水平(P <0.05或P <0.01),并且能提高小肠组织occludin和ZO-1蛋白免疫组化阳性表达(P <0.05或P <0.01);与灌胃组比较,中药灌肠组的治疗效果更为明显(P <0.05或P <0.01)。【结论】调胃承气汤可上调GOS模型大鼠肠上皮细胞紧密连接蛋白occludin、ZO-1及下调体内炎性因子TNF-α、INF-γ的表达,从而修复肠黏膜屏障,治疗GOS,且灌肠疗法要优于灌胃疗法。
Objective To observe and compare the effect of Tiaowei Chengqi Decoction by gavage and by enema on intestinal mucosal barrier of gut-origin sepsis(GOS)rat model. Methods Forty-eight rats were randomly divided into 6 groups,including sham surgery group,model group,high-and low-dose Chinese medicine gavage groups(given gavage with Tiaowei Chengqi Decoction at the dosage of 9.450,4.725 g/kg,respectively)and high-and low-dose Chinese medicine enema groups(given enema with Tiaowei Chengqi Decoction at the dosage of 9.450,4.725 g/kg,respectively), 8 rats in each group. At 2, 10, 24 h after modeling, each group was given corresponding medication, respectively. The serum levels of procalcitonin(PCT)and D-Lactate(D-LA)were detected by enzyme-linked immunosorbent assay(ELISA). The gene expression levels of occludin, zonula occluden1(ZO-1)in small intestinal tissues as well as tumor necrosis factor α(TNF-α),interferon γ(INF-γ)in liver tissues were determined by reverse transcription qPCR(RT-qPCR). The protein expression levels of occludin,ZO-1 in small intestinal tissues were observed by immunohistochemistry. And the pathological features of small intestinal tissues were observed by hematoxylin-eosin(HE)staining. Results Compared with the sham surgery group,the impair in small intestinal tissues was obvious in the model group,the serum levels of PCT and D-LA were increased(P < 0.01),the relative gene expression levels of occludin and ZO-1 in small intestinal tissues were down-regulated(P < 0.01),the relative gene expression levels of TNF-α and INF-γ in liver tissues were up-regulated(P < 0.01),and the positive immunohistochemical expression of occludin and ZO-1 in small intestinal tissues were also decreased(P < 0.05). Compared with the model group,the damage degree of small intestinal tissues was relieved in various Chinese medicine treatment groups, the serum levels of PCT and D-LA were decreased(P < 0.05 or P < 0.01),the relative gene expression levels of occludin and ZO-1 in small intestinal tissues were up-regulated,the relative gene expression levels of TNF-α and INF-γ in liver tissues were downregulated(P < 0.05 or P < 0.01),and the positive immunohistochemical expression levels of occludin and ZO-1 in small intestinal tissues were also increased(P < 0.05 or P < 0.01). And compared with the Chinese medicine gavage groups, the therapeutic effect in Chinese medicine enama groups was stronger(P < 0.05 or P < 0.01).Conclusion Tiaowei Chengqi Decoction is effective for down-regulating expression of inflammatory factors,and up-regulating expression of occludin and ZO-1 in enterocytes of GOS rats,thus to repair the barrier of intestinal mucosa to cure GOS. And the enema therapy is superior to the gavage therapy.
作者
赵锋利
吴思慧
赵馥
罗苑苑
徐运升
冼绍祥
ZHAO Feng-Li;WU Si-Hui;ZHAO Fu;LUO Yuan-Yuan;XU YurtSheng;XIAN Shao-Xiang(The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405 Guangdong, China)
出处
《广州中医药大学学报》
CAS
2019年第5期708-713,共6页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
广东省科学技术厅科技计划项目(编号:2016Z0101)