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和调健脾方对腹泻型肠易激综合征模型大鼠的治疗作用及机制研究
引用本文:陈贤家,符士颖,蔡翠珠,吕迎春.和调健脾方对腹泻型肠易激综合征模型大鼠的治疗作用及机制研究[J].陕西中医,2021,0(5):547-551,556.
作者姓名:陈贤家  符士颖  蔡翠珠  吕迎春
作者单位:(1.儋州市中医医院脾胃病科,海南 儋州 571700; 2.三亚市中医院脾胃科,海南 三亚 572000)
摘    要:目的:探讨和调健脾方对腹泻型肠易激综合征模型大鼠的治疗作用及对核因子-κB(NF-κB)/Notch1通路的影响。方法:建立腹泻型肠易激综合征大鼠模型,将造模成功的大鼠随机分为模型组、匹维溴铵组(2 mg/kg)、和调健脾方低剂量组(2.82 g/kg)、和调健脾方高剂量组(5.64 g/kg),每组12只; 另取12只健康大鼠作为对照组; 匹维溴铵组、和调健脾方低剂量组、和调健脾方高剂量组给予相应药物灌胃,对照组和模型组给予等体积蒸馏水灌胃,1次/d,连续给药14 d。给药周期结束后,检测大鼠排便频率、粪便含水量,酶联免疫吸附(ELISA)法检测大鼠血清白细胞介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)、二胺氧化酶(DAO)水平,观察大鼠结肠组织病理学变化,实时荧光定量PCR及蛋白免疫印迹法测定NF-κB、Notch1 mRNA和蛋白表达水平。结果:对照组结肠黏膜组织结构正常; 模型组肠绒毛变短、增厚、脱落,肠黏膜上皮细胞脱落,可见明显炎症细胞浸润; 匹维溴铵组及和调健脾方高剂量组结肠黏膜上皮细胞脱落减轻,炎症细胞浸润减少; 和调健脾方低剂量组仍可见明显充血及炎症细胞浸润。与对照组比较,模型组大鼠排便频率、粪便含水量、血清IL-2、TNF-α、DAO水平、结肠组织NF-κB、Notch1 mRNA和蛋白表达水平明显升高(均P<0.05); 与模型组比较,匹维溴铵组、和调健脾方低剂量组、和调健脾方高剂量组大鼠排便频率、粪便含水量、血清IL-2、TNF-α、DAO水平、结肠组织NF-κB、Notch1 mRNA和蛋白表达水平降低(均P<0.05); 且和调健脾方高剂量组大鼠排便频率、粪便含水量、血清IL-2、TNF-α、DAO水平、结肠组织NF-κB、Notch1 mRNA和蛋白表达水平低于和调健脾方低剂量组(均P<0.05); 和调健脾方高剂量组各指标与匹维溴铵组比较差异无统计学意义(均P>0.05)。结论:和调健脾方可改善腹泻型肠易激综合征模型大鼠腹泻症状,减轻大鼠血清和结肠组织炎症反应,其机制可能与和调健脾方抑制NF-κB、Notch1 mRNA和蛋白表达水平进而抑制NF-κB/Notch1通路的激活有关。

关 键 词:腹泻型肠易激综合征  和调健脾方  排便频率  粪便含水量  炎症反应  NF-κB/Notch1通路

Therapeutic effect and mechanism of Hetiao Jianpi recipe on diarrhea predominant irritable bowel syndrome model rats
CHEN Xianjia,FU Shiying,CAI Cuizhu,LYU Yingchun.Therapeutic effect and mechanism of Hetiao Jianpi recipe on diarrhea predominant irritable bowel syndrome model rats[J].Shaanxi Journal of Traditional Chinese Medicine,2021,0(5):547-551,556.
Authors:CHEN Xianjia  FU Shiying  CAI Cuizhu  LYU Yingchun
Affiliation:(Department of Spleen and Stomach Diseases,Danzhou Hospital of Traditional Chinese Medicine,Danzhou 571700,China)
Abstract:Objective:To investigate therapeutic effect of Hetiao Jianpi recipe on diarrhea predominant irritable bowel syndrome model rats and its influence on nuclear factor-κB(NF-κB)/Notch1 pathway.Methods:Establish diarrhea predominant irritable bowel syndrome model in rats,the rats were randomly divided into model group,pinaverium bromide group(2 mg/kg),Hetiao Jianpi recipe low dose group(2.82 g/kg)and Hetiao Jianpi recipe high dose group(5.64 g/kg),with 12 rats in each group; another 12 healthy rats were selected as control group; pinaverium bromide group,Hetiao Jianpi recipe low dose group and Hetiao Jianpi recipe high dose group were given corresponding drugs by gavage,while the control group and model group were given equal volume distilled water by gavage,once a day for 14 days.At the end of the administration cycle,the defecation frequency and fecal water content were detected.The serum levels of interleukin-2(IL-2),tumor necrosis factor-α(TNF-α)and diamine oxidase(DAO)were detected by enzyme-linked immunosorbent assay(ELISA).The pathological changes of colonic tissue were observed.Real time fluorescent quantitative PCR and western blotting were used to determine the levels of NF-κB,Notch1 mRNA and protein.Results:The histological structure of colonic mucosa in the control group was normal; in the model group,intestinal villi became shorter,thickened and shed,intestinal mucosal epithelial cells shed and obvious inflammatory cell infiltration could be seen; in the pinaverium bromide group and the Hetiao Jianpi recipe high dose group,colonic mucosal epithelial cells shed and inflammatory cell infiltration decreased; in the Hetiao Jianpi recipe low dose group,obvious congestion and inflammatory cell infiltration could still be seen.Compared with the control group,the defecation frequency,fecal water content,serum IL-2,TNF-α,DAO levels,colonic tissue NF-κB,Notch1 mRNA and protein expression levels in the model group were significantly increased(all P<0.05).Compared with the model group,the defecation frequency,fecal water content,serum IL-2,TNF-α,DAO levels,colonic tissue NF-κB,Notch1 mRNA and protein expression levels in the pinaverium bromide group,Hetiao Jianpi recipe low dose group and Hetiao Jianpi recipe high dose group were decreased(all P<0.05).And the defecation frequency,fecal water content,serum IL-2,TNF-α,DAO levels,colonic tissue NF-κB,Notch1 mRNA and protein expression levels in the Hetiao Jianpi recipe high dose group were lower than those in the Hetiao Jianpi recipe low dose group(all P<0.05).There was no significant difference in all indicators between Hetiao Jianpi recipe high dose group and pinaverium bromide group(all P>0.05).Conclusion:Hetiao Jianpi recipe can improve the diarrhea symptoms of diarrhea predominant irritable bowel syndrome model rats,and alleviate the inflammatory reaction of serum and colon tissue in rats.Its mechanism may be related that Hetiao Jianpi recipe inhibit NF-κB,Notch1 mRNA and protein expression levels,thereby inhibiting the activation of NF-κB/Notch1 pathway.
Keywords:Diarrhea predominant irritable bowel syndrome  Hetiao Jianpi recipe  Defecation frequency  Fecal water content  Inflammatory reaction  NF-κB/Notch1 pathway
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