文章摘要
张秉丽,霍成英,李有连,等.防风提取物通过5-羟色胺信号轴对腹泻型肠易激综合征大鼠结肠黏膜肥大细胞的影响[J].浙江中医药大学学报,2021,45(8):857-865.
防风提取物通过5-羟色胺信号轴对腹泻型肠易激综合征大鼠结肠黏膜肥大细胞的影响
Effect of Radix Saposhnikoviae Extract on Mast Cells in Colonic Mucosa of Rats with Diarrhea Predominant Irritable Bowel Syndrome through 5-hydroxytryptamine Signaling Axis
DOI:10.16466/j.issn1005-5509.2021.08.008
中文关键词: 腹泻型肠易激综合征  防风提取物  结肠黏膜肥大细胞  5-羟色胺  5-羟色胺3受体  5-羟色胺4受体  色氨酸羟化酶1  作用机制
英文关键词: diarrhea predominant irritable bowel syndrome  Radix Saposhnikoviae extract  mast cells in colonic mucosa  5-hydroxytryptamine  5-hydroxytryptamine 3 receptor  5-hydroxytryptamine 4 receptor  tryptophan hydroxylase 1  mechanism of action
基金项目:青海省卫生计生指导性科研课题([2015]18-17)
作者单位
张秉丽 青海仁济医院 西宁 810000 
霍成英 青海仁济医院 西宁 810000 
李有连 青海仁济医院 西宁 810000 
朱海宏 青海省人民医院 
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中文摘要:
      [目的]探讨防风提取物通过5-羟色胺(5-hydroxytryptamine,5-HT)信号轴对腹泻型肠易激综合征大鼠结肠黏膜肥大细胞的作用。[方法]60只清洁级健康SD大鼠中随机选择10只作为A组,剩余50只建立腹泻型肠易激综合征模型,建模成功46只,随机分为B组(11只)、C组(11只)、D组(12只)、E组(12只)。A组、B组灌胃5 mL蒸馏水;C组灌胃5-HT信号抑制剂昂丹司琼2.6mg/(kg·d)(溶于5 mL蒸馏水);D组灌胃防风提取物6.3mg/(kg·d)(溶于2.5 mL蒸馏水)+昂丹司琼2.6mg/(kg·d)(溶于2.5 mL蒸馏水);E组灌胃防风提取物6.3mg/(kg·d)(溶于5 mL蒸馏水)。观察各组大鼠精神心理行为变化,检测排便粪点数、直肠内玻璃小球排出时间;以苏木精-伊红(hematoxylin-eosin,HE)染色法观察结肠黏膜组织病理学改变;甲苯胺蓝染色法检测结肠黏膜肥大细胞数、阳性面积;实时荧光定量聚合酶链式反应(Real-time fluorescent quantitative polymerase chain reaction,Real-time qPCR)检测结肠组织5-HT3受体(5-HT3 receptor,5-HT3R)、5-HT4受体(5-HT4 receptor,5-HT4R)、色氨酸羟化酶1(tryptophan hydroxylase 1,TPH1)mRNA相对表达量;Western blot检测结肠组织5-HT3R、5-HT4R、TPH1蛋白相对表达量。[结果]与A组比较,B组、C组、D组、E组水平运动次数、垂直运动次数减少,而且C组<B组<D组<E组(P<0.05)。与A组比较,B、C、D、E组粪点数增多,直肠内玻璃小球排出时间减少,且粪点数,E组<D组<B组<C组,直肠内玻璃小球排出时间,C组<B组<D组<E组(P<0.05)。HE染色显示,与B、C组比较,D、E组结肠黏膜上皮较完整,腺体排列较整齐,间质未见炎性细胞、红细胞,其中E组改善更显著。与A组比较,B、C、D、E组肥大细胞数、阳性面积增高,E组<D组<B组<C组(P<0.05)。与A组比较,B、C、D、E组5-HT3R、TPH1 mRNA及蛋白相对表达量升高,5-HT4R mRNA及蛋白相对表达量降低,且5-HT3R、TPH1 mRNA及蛋白相对表达量,E组<D组<B组<C组,5-HT4R mRNA及蛋白相对表达量,C组<B组<D组<E组(P<0.05)。[结论]防风提取物可减少腹泻型肠易激综合征大鼠结肠黏膜肥大细胞数,改善肥大细胞功能及大鼠行为,作用机制可能与调节5-HT信号轴有关。
英文摘要:
      [Objective]To investigate the effect of Radix Saposhnikoviae extract on mast cells in colonic mucosa of rats with diarrhea predominant irritable bowel syndrome(IBS) through 5-hydroxytryptamine(5-HT) signaling axis. [Methods] Ten of sixty clean SD rats were randomly selected as group A, the remaining 50 diarrhea predominant IBS models were established, 46 were successfully modeled, and they were randomly divided into groups B(11 rats), group C(11 rats), group D(12 rats), and group E(12 rats). Group A and group B were intragastrically administered with 5 mL distilled water, group C was intragastrically administered with 5-HT signal inhibitor Ondansetron 2.6 mg/(kg·d)(dissolved in 5 mL of distilled water), group D was intragastrically administered with Radix Saposhnikoviae extract 6.3 mg/(kg·d)(dissolved in 2.5 mL of distilled water)+Ondansetron 2.6mg/(kg·d)(dissolved in 2.5 mL of distilled water), group E intragastrically Radix Saposhnikoviae extract 6.3mg/(kg·d)(dissolved in 5 mL of distilled water). The changes of mental, psychological and behavior of rats in each group were observed, and the number of defecation feces and the time of intrarectal vitreous bulb excretion were detected. Hematoxylin-eosin(HE) staining was used to observe the histopathology of colonic mucosa. The number and positive area of mast cells in colonic mucosa were detected by toluidine blue staining. The relative expression of 5-HT3 receptor(5-HT3R), 5-HT4 receptor(5-HT4R) and tryptophan hydroxylase 1(TPH1) mRNA and protein in colon tissue were detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time qPCR) and Western blot. [Results] Compared with group A, the number of horizontal and vertical movements in groups B, C, D and E decreased, and group C<group B<group D<group E(P<0.05). Compared with group A, the number of feces in groups B, C, D and E increased, and the time of intrarectal vitreous bulb excretion decreased, and for the number of feces, group E<group D<group B<group C; for the time of intrarectal vitreous bulb excretion, group C<group B<group D<group E(P<0.05). HE staining showed that compared with groups B and C, the colonic mucosa epithelium of groups D and E was more complete, the glands were arranged more orderly, no inflammatory cells and red blood cells were found in the stroma, and the improvement of group E was more significant. Compared with group A, the number and positive area of mast cells in groups B, C, D and E were increased, group E<group D<group B<group C(P<0.05). Compared with group A, the relative expressions of 5-HT3R and TPH1 mRNA and protein in groups B, C, D and E were increased, group E<group D<group B<group C; while the relative expressions of 5-HT4R mRNA and protein were decreased, group C<group B<group D<group E(P<0.05). [Conclusion] Radix Saposhnikoviae extract can reduce the number of mast cells in colon mucosa of rats with diarrhea predominant IBS, improve mast cell function and rat behavior, and the mechanism may be related to regulating the 5-HT signaling axis.
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