枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-<italic style="font-style: italic">γ</italic><sub>1</sub>/PLC-<italic style="font-style: italic">γ</italic><sub>2</sub>信号通路的影响

王伟松; 夏旭婷; 刘富林; 王文凤; 陈雨波; 滕广飞; 王婷; 涂琴蓉

doi:10.13422/j.cnki.syfjx.20191039

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枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ₁/PLC-γ₂信号通路的影响

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- 枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ₁/PLC-γ₂信号通路的影响
- Effect of Zhizhuwan on Intestinal Conduction and PLC-γ₁/PLC-γ₂ Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome
- 中国实验方剂学杂志 2019年25卷第10期页码：8-14
- 作者机构：
  
  1.湖南中医药大学　第一附属医院，长沙　 410000；
  2.湖南中医药大学　中医学院，长沙　 410000；
  3.湖南中医药大学　招生就业处，长沙　 410000；
  4.湖南中医药大学　第二附属医院，长沙　 410000
- 作者简介：
  
  王伟松，在读博士，从事消化及心血管系统疾病中医药防治研究，E-mail：wwsdipan@126.com
  *夏旭婷，硕士，助教，从事消化系统疾病中医药防治研究，E-mail：308211635@qq.com
- 基金信息：
  
  国家自然科学基金青年基金项目(81603597)
- DOI：10.13422/j.cnki.syfjx.20191039
  中图分类号： R289; R318.14; R256.35; R442.2; R285.5
- 收稿日期：2018-11-01，
  
  网络出版日期：2019-02-18，
  
  纸质出版日期：2019-05-20
- 稿件说明：
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王伟松, 夏旭婷, 刘富林, 等. 枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ₁/PLC-γ₂信号通路的影响[J]. 中国实验方剂学杂志, 2019,25(10):8-14.

Wei-song WANG, Xu-ting XIA, Fu-lin LIU, et al. Effect of Zhizhuwan on Intestinal Conduction and PLC-γ₁/PLC-γ₂ Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(10): 8-14.
王伟松, 夏旭婷, 刘富林, 等. 枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ₁/PLC-γ₂信号通路的影响[J]. 中国实验方剂学杂志, 2019,25(10):8-14. DOI： 10.13422/j.cnki.syfjx.20191039.

Wei-song WANG, Xu-ting XIA, Fu-lin LIU, et al. Effect of Zhizhuwan on Intestinal Conduction and PLC-γ₁/PLC-γ₂ Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(10): 8-14. DOI： 10.13422/j.cnki.syfjx.20191039.

摘要

目的：

探讨枳术丸对脾虚证慢传输型便秘(STC)小鼠肠道传导功能的干预作用，及对小鼠结肠组织磷脂酶C-

(PLC-

)/磷脂酶C-

(PLC-

)信号通路表达的影响。

方法：

将无特定病原体(SPF)健康小鼠随机分为正常组与造模组。采用番泻叶灌胃造成脾虚状态，然后控制饮食饮水的方法造成脾虚便秘的小鼠模型，造模完成后将模型组小鼠随机分为模型组、枳术丸组、莫沙必利组，枳术丸组按9.0 g·kg

－1

·d

－1

给药，莫沙必利组按2.5 mg·kg

－1

·d

－1

给药，模型组和正常组给予等剂量的蒸馏水，连续给药7 d，治疗完成后用印度墨水在结肠中推进的长度计算小鼠肠道推进率；用

-木糖试剂盒测定小鼠血清

-木糖含量；用苏木素-伊红(HE)染色观察小鼠结肠组织病理改变；采用免疫组化法及蛋白免疫印迹法(Western blot)检测小鼠结肠组织PLC-

，PLC-

蛋白的表达水平；采用实时荧光定量PCR(Real-time PCR)检测PLC-

，PLC-

mRNA表达水平。

结果：

与正常组比较，模型组小鼠肠道推进率及血清

-木糖值均显著降低(

0.01)，HE染色观察结肠黏膜明显充血、水肿，结肠组织PLC-

，PLC-

蛋白与mRNA表达水平均显著减少(

0.01)；与模型组比较，枳术丸组和莫沙必利组小鼠肠道推进率及血清

-木糖含量均显著增加(

0.01)，HE染色观察结肠黏膜充血、水肿及炎症细胞浸润改善，PLC-

，PLC-

蛋白与mRNA表达水平均显著升高(

0.01)。

结论：

枳术丸可有效促进脾虚便秘小鼠的肠道运动，改善便秘症状，其相关的作用机制可能与提高脾虚便秘小鼠结肠组织中PLC-

与PLC-

的表达有关。

Abstract

Objective:

To explore the effect of Zhizhuwan on intestinal conduction and expressions of Phospholipase C-

(PLC-

)/Phospholipase C-

(PLC-

) signaling pathway of slow transit constipation (STC) with spleen deficiency syndrome.

Method:

Special pathogen free (SPF) healthy mice were randomly divided into normal group and model making group. Folium Sennae gavage was used to induce the spleen deficiency status

and then diet and drinking water were controlled to establish the mice model of spleen deficiency constipation. After the modeling

the mice in modelling group were randomly divided into model group

Zhizhuwan group and mosapride group. Zhizhuwan group was given drug at the dose of 9.0 g·kg

-1

·d

-1

mosapride group was given 2.5 mg·kg

-1

·d

-1

model group and normal group were given the equal dose of distilled water for 7 consecutive days. At the end of the treatment

the length of Indian ink in the colon was used to calculate the intestinal propulsion rate of the mice. The

-xylose kit was used to determine the content of

-xylose in serum of mice. The hematoxylin eosin (HE) staining was used to observe the pathological changes of colon tissues in mice. The immunohistochemistry and Western blot were used to detect the expression levels of PLC-

and PLC-

proteins in colon tissues of mice. Real-time PCR was used to detect the mRNA expression levels of PLC-

and PLC-

Result:

Compared with normal group

the intestinal propulsive rate and the serum

-xylose value in model group were significantly decreased (

0.01). Obvious hyperemia and edema of colonic mucosa were observed by HE staining. The protein and mRNA expression levels of PLC-

and PLC-

in colon tissues were significantly decreased (

0.01). Compared with model group

the intestinal propulsion rate and the content of

-xylose in Zhizhuwan group and mosapride group were significantly increased (

0.01). Colonic mucosal hyperemia

edema and inflammatory cell infiltration were improved according to HE staining. The protein and mRNA expression levels of PLC-

and PLC-

were significantly increased (

0.01).

Conclusion:

Zhizhuwan can promote the intestinal movement in slow transit constipation model mouse with spleen deficiency syndrome

and alleviate the symptoms of constipation in mice. The related mechanism may be related to the increase of the expressions of PLC-

and PLC-

in colon tissues of mice with spleen deficiency and constipation.

关键词

Keywords

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枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ1/PLC-γ2信号通路的影响

Effect of Zhizhuwan on Intestinal Conduction and PLC-γ1/PLC-γ2 Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome

DOI：10.13422/j.cnki.syfjx.20191039

摘要

Abstract

关键词

Keywords

references

枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ₁/PLC-γ₂信号通路的影响

Effect of Zhizhuwan on Intestinal Conduction and PLC-γ₁/PLC-γ₂ Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome