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    白术配伍木香通过调节Cajal间质细胞自噬改善脾虚型腹泻肠易激综合征大鼠腹泻症状

    Atractylodes Macrocephala Koidz. with Aucklandia Lappa Decne. Improves Diarrhea Symptoms in Irritable Bowel Syndrome of Spleen Deficiency with Diarrhea Rats by Regulating Autophagy in Cajal Interstitial Cells

      摘要
    • 摘要:
      目的 探究白术配伍木香对脾虚型腹泻肠易激综合征(irritable bowel syndrome with diarrhea，IBS-D)大鼠的作用及机制。
      方法 使用液相质谱检测白术配伍木香水煎液的主要成分。以SD大鼠为研究载体，随机分为对照组、模型组、阳性对照组及低、中、高剂量白术配伍木香(n=6)。通过灌胃番泻叶联合慢性应激法建立脾虚型IBS-D模型大鼠。记录各组大鼠的一般状况及体质量变化。检测粪便含水量，肠道推进率并通过腹部撤退反射(abdominal withdrawal reflex，AWR)阈值测定评估内脏敏感性。采集大鼠结肠组织，通过透射电镜观察Cajal间质细胞(Cajal interstitial cells，ICC)内自噬小体的形成。 Western blotting检测自噬相关蛋白表达水平。免疫荧光染色观察ICC自噬水平。
      结果 模型组大鼠腹泻严重，肠道运动加快，敏感性提高。与模型组对比，白术配伍木香的治疗改善了脾虚型IBS-D大鼠的腹泻症状，恢复了大鼠体质量增长趋势，粪便含水量、肠道推进率及肠道敏感性均显著降低。治疗组大鼠结肠组织ICC内自噬小体形成数量大于模型组，自噬相关蛋白表达水平高于模型组，而干细胞因子(stem cell factor，SCF)及酪氨酸蛋白激酶kit(C-kitproto-oncogeneprotein，C-kit)表达水平低于模型组。并且，白术配伍木香对IBS-D大鼠的影响呈剂量依赖性，中剂量及高剂量白术配伍木香的作用具有显著性。
      结论 白术配伍木香通过调节ICC自噬降低脾虚型IBS-D大鼠的肠道推进率及肠道敏感性，从而改善脾虚型IBS-D的腹泻症状。

       

      Abstract:
      OBJECTIVE To investigate the effects and mechanisms of Atractylodes macrocephala Koidz. with Aucklandia lappa Decne. on irritable bowel syndrome with diarrhea(IBS-D) with spleen deficiency type in rats.
      METHODS Liquid chromatography mass spectrometry was used to detect the main components of decoction of Atractylodes macrocephala Koidz. with Aucklandia lappa Decne. SD rats were used as the study vehicle and randomly divided into control group, model group, positive control group and low-, medium-, high-dose of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. group(n=6). The rat model of spleen deficiency type IBS-D was established by intragastric administration of Cassia angustifolia Vahl combined with chronic stress method. The general conditions and weights of the rats in each group were recorded. Measured stool water content, intestinal transport rate, and visceral sensitivity by assessing the abdominal withdrawal reflex(AWR) threshold. Colonic tissue was collected from rats. The formation of autophagic vesicles in Cajal interstitial cells(ICC) was observed by transmission electron microscopy. Western blotting was used to detect the expression level of autophagy-related proteins. Immunofluorescence staining was performed to observe the level of ICC autophagy.
      RESULTS Rats in the model group showed severe diarrheal symptoms, along with accelerated intestinal motility and increased intestinal sensitivity. Compared with the model group, treatment with Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. improved the diarrheal symptoms, restored the body weight gain trend, and reduced fecal water content, intestinal transport rate, and visceral sensitivity in IBS-D rats with spleen deficiency syndrome. The number of autophagic vesicles formed in the ICC of colonic tissue in the treated rats was greater than that in the model group, and the expression levels of autophagy-related proteins were higher than those in the model group, while the expression levels of stem cell factor(SCF) and C-kitproto-oncogeneprotein(C-kit) were significantly lower than those in the model group. Moreover, the effects of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. on IBS-D rats were dose-dependent, and the effects of medium and high doses of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. were significant.
      CONCLUSION Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. reduce the intestinal transport rate and intestinal sensitivity of IBS-D rats by regulating autophagy of ICC, thereby alleviating diarrhea symptoms in IBS-D with spleen deficiency syndrome.

       

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