Effects of <italic style="font-style: italic">Qingre Huashi Tiaoshu Formula</italic> （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice

Xiaolan SU; Mengxi YAO; Lin CHEN; Tao ZHANG; Lin WANG; Xiangli QING; Gezhi ZHANG; Wei WEI

doi:10.13288/j.11-2166/r.2022.16.012

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Effects of Qingre Huashi Tiaoshu Formula （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice

更新时间：2022-09-15

- Effects of Qingre Huashi Tiaoshu Formula （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice
- Journal of Traditional Chinese Medicine Vol. 63, Issue 16, Pages: 1566-1572(2022)
- 作者机构：
  
  1.中国中医科学院望京医院，北京市朝阳区望京中环南路6号，100102
  2.中国中医科学院
  3.成都中医药大学
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2022.16.012
  CLC：
- Received：06 January 2022，
  
  Revised：2022-04-21，
  
  Published：17 August 2022
- 稿件说明：
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苏晓兰,姚梦茜,陈琳等.清热化湿调枢方对葡聚糖硫酸钠诱导的结肠炎模型小鼠肠黏膜屏障及相关蛋白表达的影响[J].中医杂志,2022,63(16):1566-1572.

SU Xiaolan,YAO Mengxi,CHEN Lin,et al.Effects of Qingre Huashi Tiaoshu Formula （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice[J].Journal of Traditional Chinese Medicine,2022,63(16):1566-1572.
苏晓兰,姚梦茜,陈琳等.清热化湿调枢方对葡聚糖硫酸钠诱导的结肠炎模型小鼠肠黏膜屏障及相关蛋白表达的影响[J].中医杂志,2022,63(16):1566-1572. DOI： 10.13288/j.11-2166/r.2022.16.012.

SU Xiaolan,YAO Mengxi,CHEN Lin,et al.Effects of Qingre Huashi Tiaoshu Formula （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice[J].Journal of Traditional Chinese Medicine,2022,63(16):1566-1572. DOI： 10.13288/j.11-2166/r.2022.16.012.

摘要

目的

探讨清热化湿调枢方治疗溃疡性结肠炎的可能作用机制。

方法

36只C57BL/6小鼠随机抽取6只小鼠为正常组，其余小鼠均给予2.5%葡聚糖硫酸钠（DSS）水溶液自由饮用7天以制备DSS结肠炎小鼠模型。将造模成功小鼠再随机分为模型组，美沙拉嗪组以及清热化湿调枢方低、中、高剂量组，每组6只。正常组和模型组小鼠予0.5 ml纯水灌胃；美沙拉嗪组小鼠给予0.52 g/（kg·d）美沙拉嗪混悬液灌胃；清热化湿调枢方低、中、高剂量组小鼠分别给予清热化湿调枢方0.845、1.690、3.380 g/（kg·d）灌胃；各组均每日1次，连续10天。干预后测量各组小鼠体质量并进行疾病活动指数（DAI）评分，测量结肠长度并观察结肠组织病理变化，检测结肠组织黏蛋白2（MUC-2）阳性表达及带状闭合蛋白1（ZO-1）、闭合蛋白1（Claudin-1）、咬合蛋白（Occludin）表达。

结果

与正常组相比，模型组和清热化湿调枢方低剂量组小鼠的体质量下降，DAI评分明显升高，结肠长度变短，结肠组织ZO-1、Occludin、Claudin-1蛋白表达均降低（

＜0.05）。与模型组和清热化湿调枢方低剂量组比较，美沙拉嗪组和清热化湿调枢方高、中剂量组小鼠体质量增加，DAI评分下降，结肠长度增长，结肠组织ZO-1、Claudin-1、Occludin蛋白表达升高（

＜0.05）。正常组小鼠肠黏膜上皮完整，无炎性细胞浸润；模型组小鼠肠黏膜上皮缺损，杯状细胞明显减少，固有层及隐窝结构破坏严重，排列紊乱，并可见大量淋巴细胞浸润；清热化湿调枢方高剂量组和美沙拉嗪组小鼠结肠黏膜上皮几乎无缺损，极少炎性细胞浸润；而清热化湿调枢方中、低剂量组改善不明显。正常组MUC-2表达为棕黄色颗粒，广泛分布在杯状细胞胞浆（核周及核膜）；模型组小鼠结肠组织中MUC-2阳性表达明显减少；美沙拉嗪组和清热化湿调枢方高、中剂量组MUC-2阳性表达较模型组有较明显的恢复。

结论

高、中剂量清热化湿调枢方对DSS结肠炎小鼠的肠道黏膜屏障损伤具有一定修复作用，其作用机制可能是增加结肠组织MUC-2表达，促进ZO-1、Occludin、Claudin-1分泌，从而促进黏膜化学及机械屏障的修复，维持肠黏膜屏障的稳态。

Abstract

Objective

To explore the possible mechanism of

Qingre Huashi Tiaoshu Formula

（清热化湿调枢方， QHTF） in the treatment of ulcerative colitis （UC）.

Methods

Thirty-six C57BL/6 male mice were included， of which six mice were randomly selected as normal group， and the rest were given 2.5% dextran sodium sulfate （DSS） aqueous solution for 7 days to prepare DSS-induced colitis mice. The successfully modeled mice were randomly divi-ded into five groups， namely the model group， mesalazine group， and the low-， medium- and high-dose QHTF groups， with six mice in each group.Then the normal group and the model group were given 0.5 ml of pure water by gavage； mice in the mesalazine group were given 0.52 g/（kg·d） of mesalazine suspension by gavage； mice in the low-， medium- and high-dose QHTF groups were given 0.845， 1.69， and 3.38 g/（kg·d） of QHTF suspension by gavage. All groups were intervented once a day for 10 consecutive days. After intervention， the body weight， disease activity index （DAI）， colon length， colon histopathological results were observed， and the positive expression of colonic tissue mucin 2 （MUC-2） and the expression of occludin 1 （ZO-1）， claudin 1 （Claudin-1） and occludin （Occludin） were detected.

Results

Compared to those of mice in the normal group， the body weight of mice in the model group and low-dose QHTF group decreased， and the DAI score increased， while colon length was shortened， and the expression of ZO-1， Occludin， and Claudin-1 in the colonic tissue decreased （

＜0.05）. Compared to the model group and low-dose QHTF group， the other three groups had increased body weight， decreased DAI， longer colon length， and increased expression of ZO-1， Occludin， and Claudin-1 （

＜0.05）. In the normal group， the intestinal mucosal epithelium of the mice was intact with no inflammatory cell infiltration， while in the model group， the intestinal mucosal epithelium was defective with significantly reduced goblet cells， severely damaged and disorderly arranged lamina propria and crypts structure， and a large number of infiltrated lymphocytes； in the high-dose QHTF group and mesalazine group， the colonic mucosal epithelium had almost no defect and very little inflammatory cell infiltration， while in the middle- and low-dose QHTF groups， it was not significantly improved. The MUC-2 was expressed in brown-yellow granules， widely distributed in the cytoplasm （perinuclear and nuclear membrane） in the normal group； the positive expression of MUC-2 was significantly reduced in the model group； in the mesalazine group and the high- and medium-dose QHTF groups， the positive expression of MUC-2 was significantly restored compared to that in the model group.

Conclusion

High- and medium-dose QHTF have certain effect on repairing the intestinal mucosal barrier damage in DSS colitis mice， and the mechanism may be related to its action on increaing the expression of MUC-2 in the colonic tissue， promoting the secretion of ZO-1， Occludin， and Claudin-1， thereby promoting the repair of mucosal chemical and mechanical barriers， and maintaining the homeostasis of intestinal mucosal barrier.

关键词

Keywords

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Effects of Qingre Huashi Tiaoshu Formula （清热化湿调枢方） on Intestinal Mucosal Barrier and Expression of Related Proteins in Dextran Sodium Sulfate-induced Colitis Model Mice

DOI：10.13288/j.11-2166/r.2022.16.012

摘要

Abstract

关键词

Keywords

references