中药复方通腑颗粒及其组分对脓毒症大鼠肠黏膜机械屏障的影响

目的 探讨中药复方通腑颗粒及其组分对脓毒症大鼠肠黏膜机械屏障的保护作用.方法 按随机数字表法将大鼠分为模型组、大黄组、厚朴组、通腑颗粒组,以正常大鼠作为正常对照组.腹腔注射大肠杆菌脂多糖(LPS)6 mg/kg制备大鼠脓毒症模型,制模成功后通腑颗粒组灌胃通腑颗粒28 g·kg-1·d-1,大黄组、厚朴组分别灌胃大黄、厚朴5 g·kg-1·d-1,模型组灌胃等量生理盐水,各组均每日1次.分别于术后24、48及72 h心脏取血,采用酶联免疫吸附法(ELISA)测定血清肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)水平;72 h时取小肠组织,观察小肠黏膜病理改变并进行病理评分.结果 模型组各时间点血清TNF-α、IL-8均较正常对照组明显升高;各治疗组治疗后血清TNF-α、IL-8均明显低于模型组;通腑颗粒组各时间点TNF-α(ng/L)明显低于大黄、厚朴组(24 h:44.64±1.48比47.18±1.83和46.96±2.23,48 h:51.38±1.36比57.17±2.23和59.41±2.01,72 h:55.54±2.58比64.34±1.02和65.96±3.45,均P<0.05);72 h时IL-8(ng/L)明显低于厚朴组(65.53±4.52比69.14±2.82,P<0.05).正常对照组小肠黏膜病理评分为0;大黄、厚朴、通腑颗粒组小肠黏膜病理评分(分)均明显低于模型组(3.15±0.28、 3.18±0.08、 2.95±0.15比3.90±0.17,均P<0.01),且以通腑颗粒组下降程度明显.电镜下观察正常对照组小肠黏膜紧密连接完整,微绒毛及线粒体形态正常;模型组小肠黏膜微绒毛缺失、崩解,紧密连接破坏;大黄组与厚朴组小肠黏膜细胞间紧密连接局部模糊;通腑颗粒组小肠黏膜细胞间紧密连接保存相对完整.结论 脓毒症大鼠存在肠黏膜屏障损伤;中药通腑颗粒及其组分大黄、厚朴均可降低脓毒症大鼠血清TNF-α和IL-8水平,减轻肠黏膜损伤,从而起到保护肠道屏障的作用,以通腹颗粒作用明显.

Abstract：

Objective To investigate the effects of Tongfu granules (通腑颗粒) and its constituents on barrier function of small intestine in rats with sepsis. Methods The male rats were divided into model group, Tongfu granules group, Rhubarb (大黄) group and Magnoliae cortex (厚朴) group by random digits table, normal rats as control group. Intraperitoneal injection of lipopolysaccharide (LPS, 6 mg/kg) was used to reproduce sepsis model. After establishment of model, rats in Tongfu granules group were given Tongfu granules 28 g·kg-1·d-1 by gavage, and Rhubarb group and Magnoliae cortex group rats were given Rhubarb or Magnoliae cortex 5 g·kg-1·d-1 by gavage, while the model group was given normal saline in same quantity, once a day. Blood samples of rats were collected at 24, 48, 72 hours for measuring tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) with enzyme linked immunosorbent assay (ELISA). The pathological changes in intestinal mucosa were observed, and the pathological scores was estimated at 72 hours. Results The levels of TNF-α and IL-8 were significantly higher in model group than those in control group at different time points. The serum levels of TNF-α and IL-8 were significantly lower in treatment groups than those in model group, and the level of TNF-α (ng/L) in Tongfu granules group was significantly lower than that in Rhubarb and Magnoliae cortex groups at different time points (24 hours: 44.64±1.48 vs. 47.18±1.83 and 46.96±2.23, 48 hours: 51.38±1.36 vs. 57.17±2.23 and 59.41±2.01, 72 hours: 55.54±2.58 vs. 64.34±1.02 and 65.96±3.45, all P<0.05), and IL-8 (ng/L) level at 72 hours was significantly lower than that in Magnoliae cortex group (65.53±4.52 vs. 69.14±2.82, P<0.05). The scores of the lesions were significantly higher in model group than that in control group (3.90±0.17 vs. 0). The scores of Rhubarb group, Magnoliae cortex group and Tongfu granules group were 3.15±0.28, 3.18±0.08, and 2.95±0.15, respectively, which were lower than those of the model group (all P<0.01), and the Tongfu granules group descended obviously than other groups. In control group, the intercellular tight junctions were normal, and the morphology of microvilli and mitochondria was also normal. In model group, the microvilli of intestinal mucosa of the small intestine were absent or disintegrated. The intercellular tight junctions were seen to be blurred in Rhubarb and Magnoliae cortex groups, and they were close to normal state in Tongfu granules group. Their integrity was better preserved compared with that of the model group. Conclusion Injury of barrier function of the small intestine was found in septic rat. It was found that traditional Chinese medicine Tongfu granules, Rhubarb and Magnoliae cortex could protect the barrier function of the small intestine by decreasing the TNF-α and IL-8 levels in septic rats. Above-mentioned effects of Tongfu granules were better than Rhubarb and Magnoliae cortex.