Effect of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and interstitial cells of Cajal in mice with slow transit constipation

Chen Rong; Zhou Biying; Zhou Ling

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Effect of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and interstitial cells of Cajal in mice with slow transit constipation

更新时间：2022-06-24

- Effect of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and interstitial cells of Cajal in mice with slow transit constipation
- Journal of Chongqing Medical University Vol. 47, Issue 1, Pages: 1-8(2022)
- 作者机构：
  
  1. 遵义医科大学附属医院康复医学科,遵义,563000
  2. 遵义医科大学寄生虫学教研室,遵义,563000
  3. 遵义医科大学附属医院中医科,遵义,563000
- 作者简介：
- 基金信息：
- DOI：
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- Published：2022，
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Chen Rong, Zhou Biying, Zhou Ling. Effect of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and interstitial cells of Cajal in mice with slow transit constipation. [J]. Journal of Chongqing Medical University 47(1):1-8(2022)
DOI：

Chen Rong, Zhou Biying, Zhou Ling. Effect of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and interstitial cells of Cajal in mice with slow transit constipation. [J]. Journal of Chongqing Medical University 47(1):1-8(2022) DOI：

摘要

目的：探究白术破壁饮片对慢传输型便秘（slow transit constipation，STC）小鼠肠神经递质、Cajal间质细胞（interstitial cells of Cajal，ICC）的干细胞因子（stem cell factor，SCF）/干细胞生长因子受体（c-kit）信号通路的影响。方法：采用皮下注射2.5 mg/（kg·d）盐酸吗啡注射液法复制STC小鼠模型，选取48只雌雄各半昆明小鼠随机分为正常组，模型组，白术破壁饮片低、中、高剂量组（25、50、100 mg/kg）和莫沙必利组（1.5 mg/kg），每组8只。各组予以不同药物干预1周后，检测造模后和给药后各组小鼠粪便性状、肠道推进率；采用酶联免疫反应（enzyme-linked immunosorbent assay，ELISA）测定各组小鼠结肠神经递质P物质（substance P，SP）、乙酰胆碱（acetylcholine，Ach）、血管活性肠肽（vasoactive intestinal peptide，VIP）、一氧化氮（nitric oxide，NO）和5-羟色胺（5-hydroxytrytamine，5-HT）含量；采用Western blot检测小鼠结肠组织SCF和c-kit蛋白的表达。结果：ELISA结果显示，与正常组比较，模型组SP和Ach含量均明显降低（SP：230.41±16.41 vs. 146.69±15.59；Ach：577.68±39.35 vs. 281.50±39.40）。与模型组比较，白术破壁饮片低剂量组、白术破壁饮片中剂量组和白术破壁饮片高剂量组SP含量均明显升高（146.69±15.59 vs. 188.25±4.60；146.69±15.59 vs. 201.78±4.44；146.69±15.59 vs. 307.51±16.94）；白术破壁饮片高剂量组和莫沙必利组Ach含量均明显升高（281.50±39.40 vs. 400.93±12.21；281.50±39.40 vs. 422.22±22.70）。与白术破壁饮片高剂量组比较，白术破壁饮片低剂量组和白术破壁饮片中剂量组SP和Ach含量均明显降低（SP：307.51±16.94 vs.188.25±4.60；307.51±16.94 vs. 201.78±4.44；Ach：400.93±12.21 vs. 283.79±11.53；400.93±12.21 vs. 327.81±14.77）；莫沙必利组SP含量均明显降低（307.51±16.94 vs. 164.08±12.54）。与正常组比较，模型组VIP、NO和5-HT含量均明显升高（VIP： 64.47±2.69 vs. 87.74±2.93；NO：38.21±1.76 vs. 42.78±1.69；5-HT：219.58±11.60 vs. 276.08±7.97）。与模型组比较，白术破壁饮片低剂量组、白术破壁饮片中剂量组、白术破壁饮片高剂量组和莫沙必利组VIP含量均明显降低（87.74±2.93 vs. 75.58±2.08；87.74±2.93 vs. 69.34±2.23；87.74±2.93 vs. 66.37±1.93；87.74±2.93 vs. 65.31±3.32 ）；白术破壁饮片中剂量组、白术破壁饮片高剂量组和莫沙必利组NO和5-HT含量均明显降低（NO：42.78±1.69 vs. 39.27±1.90；42.78±1.69 vs. 37.43±1.30；42.78±1.69 vs. 35.65±2.01；5-HT：276.08±7.97 vs. 257.89±6.16；276.08±7.97 vs. 226.79±10.49；276.08±7.97 vs. 242.05±12.15）。与白术破壁饮片高剂量组比较，白术破壁饮片低剂量组VIP、NO和5-HT含量均明显升高（VIP：66.37±1.93 vs. 75.58±2.08；NO：37.43±1.30 vs. 42.73±2.19；5-HT：226.79±10.49 vs. 269.87±10.91）；白术破壁饮片中剂量组VIP、5-HT，莫沙必利组5-HT含量均明显升高。Western blot结果显示，与模型组比较，白术破壁饮片低剂量组c-kit蛋白含量明显升高（0.22±0.10 vs. 0.37±0.08）；白术破壁饮片中剂量组、白术破壁饮片高剂量组和莫沙必利组SCF和c-kit蛋白含量均明显升高（SCF：0.60±0.19 vs. 0.99±0.28；0.60±0.19 vs. 1.17±0.34；0.60±0.19 vs. 1.02±0.30；c-kit：0.22±0.10 vs. 0.47±0.10；0.22±0.10 vs. 0.58±0.13；0.22±0.10 vs. 0.49±0.13）。结论：白术破壁饮片可通过升高STC模型小鼠结肠组织 SP、Ach和SCF/c-kit含量，以及降低VIP、NO和5-HT含量，从而调节小鼠胃肠道功能，促进肠道蠕动，改善便秘症状，进而达到治疗STC的效果。

Abstract

Objective：To investigate the effects of Atractylodes macrocephala decoction pieces on enteric neurotransmitter and stem cell factor（SCF）/stem cell growth factor receptor（c-kit） signaling pathway of interstitial cells of Cajal（ICC） in mice with slow transit constipation（STC）. Methods：The STC mouse model was replicated by subcutaneous injection of 2.5 mg/（kg·d） morphine hydrochloride injection. A total of 48 Kunming mice were randomly divided into normal group，model group，Atractylodes macrocephala decoction pieces low，medium，high-dose groups（25，50，100 mg/kg） and mosapride group（1.5 mg/kg），each with 8 mice. After each group was treated with different drugs for 1 week，the stool characteristics and intestinal propulsion rate of mice in each group were tested after modeling and administration. Enzyme-linked immunosorbent assay（ELISA） was conducted to determine neurotransmitter substance P（SP），acetylcholine（Ach），vasoactive intestinal peptide（VIP），nitric oxide（NO） and 5-hydroxytrytamine（5-HT） contents in the colon of the mice in each group. And Western blot was used to detect the expression of SCF and c-kit protein in mouse colon tissue. Results：ELISA results showed that compared with the normal group，the SP and Ach contents of the model group were significantly reduced（SP：230.41±16.41 vs. 146.69±15.59；Ach：577.68±39.35 vs. 281.50±39.40）. Compared with the model group，the SP content in the low-dose group of Atractylodes macrocephala，the middle-dose group and the high-dose group of Atractylodes macro－cephala fragments were significantly increased（146.69±15.59 vs. 188.25±4.60；146.69±15.59 vs. 201.78±4.44；146.69±15.59 vs. 307.51±16.94）；Ach content in high-dose Atractylodes macrocephala and mosapride group were significantly increased（281.50±39.40 vs. 400.93±12.21；281.50±39.40 vs. 422.22±22.70 ）. Compared with the high-dose group of Atractylodes macrocephala decoction pieces，the SP and Ach contents of the low-dose group of Atractylodes macrocephalus decoction pieces and the middle-dose group of Atractylodes macrocephalus decoction pieces were significantly reduced（SP：307.51±16.94 vs. 188.25±4.60；307.51±16.94 vs. 201.78±4.44；Ach：400.93±12.21 vs. 283.79±11.53；400.93±12.21 vs. 327.81±14.77）；SP content in the mosapride group was significantly reduced（307.51±16.94 vs. 164.08±12.54）. Compared with the normal group，the contents of VIP，NO and 5-HT in the model group were significantly increased（VIP：64.47±2.69 vs. 87.74±2.93；NO：38.21±1.76 vs. 42.78±1.69；5-HT：219.58±11.60 vs. 276.08±7.97）. Compared with the model group，the VIP content of the low-dose Atractylodes macrocephala decoction pieces，the medium-dose Atractylodes macrocephalus decoction pieces，the high-dose Atractylodes macrocephalus decoction pieces，and the mosapride group were significantly reduced（87.74±2.93 vs. 75.58±2.08；87.74±2.93 vs. 69.34±2.23；87.74±2.93 vs. 66.37±1.93；87.74±2.93 vs. 65.31±3.32）；Atractylodes macrocephala medium dose group，Atractylodes macrocephala fragment high dose group and mosapride group NO and The content of 5-HT was significantly reduced（NO：42.78±1.69 vs. 39.27±1.90；42.78±1.69 vs. 37.43±1.30；42.78±1.69 vs. 35.65±2.01；5-HT：276.08±7.97 vs. 257.89±6.16；276.08±7.97 vs. 226.79±10.49；276.08±7.97 vs. 242.05±12.15）. Compared with the high-dose Atractylodes macrocephala decoction pieces group，the contents of VIP，NO and 5-HT in the low-dose Atractylodes macrocephalus decoction pieces group were significantly increased（VIP：66.37±1.93 vs. 75.58±2.08；NO：37.43±1.30 vs. 42.73±2.19；5-HT：226.79±10.49 vs. 269.87±10.91）；VIP，5-HT in the middle-dose group of Atractylodes macrocephala and 5-HT in the mosapride group were significantly increased. Western blot results showed that compared with the model group，the c-kit protein content in the low-dose group of Atractylodes macrocephala was significantly increased（0.22±0.10 vs. 0.37±0.08）；the middle-dose group of Atractylodes macrocephala and the high-dose group of Atractylodes macrocephala both the SCF and c-kit protein content in the mosapride group and the mosapride group were significantly increased（SCF：0.60±0.19 vs. 0.99±0.28；0.60±0.19 vs. 1.17±0.34；0.60±0.19 vs. 1.02±0.30；c-kit：0.22±0.10 vs. 0.47±0.10；0.22±0.10 vs. 0.58±0.13；0.22±0.10 vs. 0.49±0.13）. Conclusion：Atractylodes macrocephala decoction pieces can increase the contents of SP，Ach and SCF/c-kit in the colon tissue of STC model mice，and reduce the contents of VIP，NO and 5-HT，thereby regulating the gastrointestinal function of mice，promoting the intestinal tract peristalsis，improving constipation symptoms，and then achieving the effect of treating STC.

关键词

白术破壁饮片慢传输型便秘结肠神经递质Cajal间质细胞

Keywords

Atractylodes macrocephala decoction piecesslow transit constipationcolonic neurotransmitterinterstitial cells of Cajal

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