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陈镇,向彪,周鑫,付子煜,刘涛,吴金鸿,欧阳林旗,喻斌,邓桂明.不同产地乌药质量评价及其改善IBS-D大鼠肠道低度炎症的机制研究[J].湖南中医药大学学报英文版,2023,43(11):1963-1970.[Click to copy
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| 不同产地乌药质量评价及其改善IBS-D大鼠肠道低度炎症的机制研究 |
| 陈镇,向彪,周鑫,付子煜,刘涛,吴金鸿,欧阳林旗,喻斌,邓桂明 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 基于高效液相色谱法(high performance liquid chromatography, HPLC)对10批次不同产地乌药进行质量评价,并进一步探究乌药对腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome, IBS-D)大鼠肠道低度炎症的作用机制。方法 采用HPLC对10个不同批次乌药进行质量评价。选取雄性SD大鼠60只,随机分为对照组、IBS-D模型组、乌药低剂量组、乌药中剂量组、乌药高剂量组、阳性药物组。除对照组外,其余大鼠均接受"番泻叶灌胃联合束缚应激"刺激,构建IBS-D大鼠模型。乌药低、中、高剂量组大鼠分别灌胃乌药水提物(0.94、1.88、3.76 g·kg-1),阳性药物组大鼠予以匹维溴铵片水溶液(1.5 g·kg-1);而对照组和IBS-D模型组大鼠灌胃等体积蒸馏水,共计2周;采用HE染色、PAS染色观察大鼠结肠变化;运用免疫组织化学法和Western blot技术检测大鼠结肠组织中过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ, PPAR-γ)、血管细胞黏附分子1(vascular cell adhesion molecular-1, VCAM-1)、原癌基因酪氨酸蛋白激酶(proto-oncogene tyrosine-protein kinase Src, SRC)、Yes相关蛋白(Yes-associated protein, YAP)蛋白的相对表达水平。结果 相似度评价、聚类结果和主成分分析结果显示,除浙江乌药(S6)外,其余批次乌药质量从整体上而言稳定可靠。HE染色和PAS染色结果显示,与对照组相比,IBS-D模型组大鼠结肠隐窝深度明显变浅、隐窝处杯状细胞数量明显减少;与IBS-D模型组相比,中、高剂量的乌药水提物能明显改善IBS-D大鼠结肠组织隐窝深度变浅及杯状细胞减少的状态;免疫组织化学和Western blot结果显示,3个剂量组的乌药水提物均可显著增强IBS-D大鼠结肠中PPAR-γ、YAP和SRC蛋白的相对表达量(P＜0.05),同时能显著降低VCAM-1蛋白的相对表达量(P＜0.05)。结论 不同批次的乌药化学成分和含量差异不大,乌药能显著改善IBS-D大鼠低度炎症,其作用机制可能与调控PPAR-γ/VCAM-1有关。 |
| 关键词: 乌药 腹泻型肠易激综合征 质量评价 低度炎症 PPAR-γ/VCAM-1信号通路 作用机制 |
| DOI：10.3969/j.issn.1674-070X.2023.11.005 |
| Received:September 04, 2023 |
| 基金项目:国家自然科学基金面上项目（82374424）；湖南省自然科学基金项目（2018JJ6040，2020JJ5434，2022JJ70028）；湖南省中医药科研计划项目（2021016）；湖南省教育厅科学研究项目（19A372）；湖南中医药大学科研基金项目（2022YYZK002，2019XJJJ025）；湖南中医药大学中药学重点学科。 |
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| Quality evaluation of Wuyao from different origins and its mechanism in alleviating intestinal low-grade inflammation in IBS-D rats |
| CHEN Zhen,XIANG Biao,ZHOU Xin,FU Ziyu,LIU Tao,WU Jinhong,OUYANG Linqi,YU Bin,DENG Guiming |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To evaluate the quality of 10 batches of Wuyao (Linderae Radix) from different origins based on high performance liquid chromatography (HPLC), and to further explore its mechanism of action on intestinal low-grade inflammation in rats with diarrhea-predominant irritable bowel syndrome (IBS-D). Methods The quality of 10 different batches of Wuyao (Linderae Radix) was evaluated by HPLC. Sixty male SD rats were selected and assigned randomly into control group, IBS-D model group, low-, medium-, and high-dose Wuyao (Linderae Radix) groups, and positive drug group. Except the control group, all the other rats were stimulated by "Fanxieye (Sennae Folium) gavage combined with restraint stress" to construct IBS-D rat model. The low-, medium-, and high-dose Wuyao (Linderae Radix) groups were given intragastric administration of Wuyao (Linderae Radix) water extract (0.94, 1.88, 3.76 g·kg-1), and the positive drug group was given aqueous solution of pinaverium bromide tablets (1.5 g·kg-1). The control group and IBS-D model group were treated with equal volume of distilled water by gavage for 2 weeks. HE and PAS staining were used to observe the colonic changes respectively. The relative expression levels of peroxisome proliferator-activated receptor γ (PPAR-γ), vascular cell adhesion molecule-1 (VCAM-1), proto-oncogene tyrosine-protein kinase Src (SRC), and Yes-associated protein (YAP) in rat colon tissues were determined by immunohistochemistry and Western blot. Results Similarity evaluation, clustering results, and principal component analysis showed that the quality of other batches was stable and reliable overall except Zhejiang Wuyao (Linderae Radix) (S6). HE staining and PAS staining showed that compared with the control group, the depth of colonic crypts in the IBS-D model group was significantly shallower and there was a noticeable decrease in the number of goblet cells in the crypts. Compared with IBS-D model group, the medium and high doses of Wuyao (Linderae Radix) water extract demonstrated significant improvement in the depth of crypt of colon tissues in IBS-D rats and the reduction of goblet cells. Immunohistochemistry and Western blot showed that Wuyao (Linderae Radix) water extract of the three dose groups could significantly enhance the relative expressions of PPAR-γ, YAP, and SRC in the colon of IBS-D rats (P＜0.05), and also significantly decreased the relative expression of VCAM-1 protein (P＜0.05). Conclusion There was little difference in chemical composition and content for different batches of Wuyao (Linderae Radix). It can notably alleviate low-grade inflammation in IBS-D rats, and its mechanism of action may be related to the regulation of PPAR-γ/VCAM-1 signaling pathways. |
| Key words: Wuyao (Linderae Radix) diarrhea-predominant irritable bowel syndrome quality evaluation low-grade inflammation peroxisome proliferator-activated receptor γ/Vascular cell adhesion molecule-1 signaling pathway mechanism of action |
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