白术配伍木香通过调节Cajal间质细胞自噬改善脾虚型IBS-D大鼠腹泻症状

Atractylodes macrocephala Koidz. with Aucklandia lappa Decne. improves diarrhea symptoms in IBS-D rats by regulating autophagy in Cajal interstitial cells

关键词:  腹泻型肠易激综合征  白术  木香  Cajal间质细胞  自噬

Key words:  Irritable bowel syndrome with diarrhea  Atractylodes macrocephala Koidz.  Aucklandia lappa Decne.  Cajal interstitial cells  Autophagy

摘要:

目的：探究白术配伍木香对脾虚腹泻型肠易激综合征(脾虚型IBS-D)大鼠的作用及机制。方法：使用液相质谱检测白术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、低剂量白术-木香组、中剂量白术-木香组、高剂量白术-木香组及阳性对照组(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重变化。检测粪便含水量，肠道推进率并通过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(ICC)内自噬小体的形成。WB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，恢复了大鼠体重增长趋势。粪便含水量，肠道推进率及肠道敏感性均显著低于模型组。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SCF及C-kit表达水平低于模型组。并且，白术-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-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 rats with irritable bowel syndrome of spleen deficiency type diarrhea (IBS-D). 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, low-dose of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. group, medium-dose of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. group, high-dose of Atractylodes macrocephala Koidz. and Aucklandia lappa Decne. group and positive group (n=6). Rats were established by intragastric administration of Cassia angustifo lia Vahl combined with chronic stress method for the diarrhea-type irritable bowel syndrome model. The general conditions and weights of the rats in each group were recorded. Visceral sensitivity was assessed by AWR threshold assay and stool water content, intestinal transport rate was measured. Colonic tissue was collected from rats. The formation of autophagic vesicles in Cajal interstitial cells (ICC) was observed by transmission electron microscopy. WB 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 and restored the body weight of the rats with spleen deficiency type IBS-D. Fecal water content, intestinal transport rate and intestinal sensitivity were significantly lower than those in the model group. 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 SCF and 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. reduced the intestinal transport rate and intestinal sensitivity of IBS-D rats by regulating autophagy of Cajal interstitial cells, thereby improving the diarrheal symptoms of IBS-D.