Multi-target Regulatory Effect of Active Ingredients of Sinisan on 5-HT Signal Pathway in Visceral Hypersensitivity Rats

ZHANG Jing-xian; RUI Jun-qian; CHEN Yi-fan; MIAO Le-tao; YANG Xing-hao

doi:10.13422/j.cnki.syfjx.20181625

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Multi-target Regulatory Effect of Active Ingredients of Sinisan on 5-HT Signal Pathway in Visceral Hypersensitivity Rats

更新时间：2024-01-04

- Multi-target Regulatory Effect of Active Ingredients of Sinisan on 5-HT Signal Pathway in Visceral Hypersensitivity Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 16, Pages: 115-123(2018)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20181625
  CLC： R285.5
- Published：2018
- 稿件说明：
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ZHANG Jing-xian, RUI Jun-qian, CHEN Yi-fan, et al. Multi-target Regulatory Effect of Active Ingredients of Sinisan on 5-HT Signal Pathway in Visceral Hypersensitivity Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 115-123.
DOI：

ZHANG Jing-xian, RUI Jun-qian, CHEN Yi-fan, et al. Multi-target Regulatory Effect of Active Ingredients of Sinisan on 5-HT Signal Pathway in Visceral Hypersensitivity Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 115-123. DOI： 10.13422/j.cnki.syfjx.20181625.

摘要

目的：研究芍药苷、辛弗林、甘草酸对内脏高敏感大鼠5-羟色胺（5-HT）信号通路的多靶点[色氨酸羟化酶1（TPH1）/5-羟色胺转运体（SERT）/5羟色胺3受体（5-HT3R）/5-HT4RI调控，探讨各成分间的协同交互作用，探索四逆散有效成分干预内脏高敏感状态的潜在机制。方法： SD大鼠随机分为正常组，模型组，匹维溴铵组（15 mg·kg-1），芍药苷组（50 mg·kg-1），辛弗林组（50 mg·kg-1），甘草酸组（50 mg·kg-1），芍药苷+甘草酸组（25 mg·kg-1+25 mg·kg-1），芍药苷+辛弗林组（25 mg·kg-1+25 mg·kg-1），辛弗林+甘草酸组（25 mg·kg-1+25 mg·kg-1），芍药苷+辛弗林+甘草酸组（16.7 mg·kg-1+16.7 mg·kg-1+16.7 mg·kg-1），四逆散组（10 g·kg-1）。采用复合应激法建立肠易激综合征（IBS）内脏高敏感大鼠模型；以大鼠一般状态评分和直肠扩张腹壁撤退反射（AWR）的最小容量阈值评价大鼠肠道敏感性程度；采用酶联免疫吸附法（ELISA）检测大鼠结肠组织5-HT，TPH1，SERT，5-HT3R，5-HT4R水平，实时荧光定量聚合酶链式反应（Real-time PCR）检测大鼠结肠组织TPH1，5-HT3R，5-HT4R mRNA表达水平，蛋白免疫印迹法（Western blot）检测大鼠结肠组织TPH1，5-HT3R蛋白水平。结果：与正常组比较，模型组大鼠一般状态评分和最小容量阈值显著下降（P<0.01），结肠组织5-HT，TPH1，5-HT3R水平显著升高，SERT，5-HT4R水平明显降低（P<0.05，P<0.01），TPH1，5-HT3R mRNA及蛋白表达显著升高，5-HT4R mRNA表达明显降低（P<0.05，P<0.01）。与模型组比较，芍药苷+辛弗林+甘草酸组结肠5-HT，TPH1，5-HT3R水平明显降低，SERT，5-HT4R水平明显升高，TPH1，5-HT3R mRNA及蛋白表达明显降低，5-HT4R mRNA表达明显升高（P<0.05，P<0.01）；TPH1，5-HT3R，5-HT4R mRNA多靶点纠偏作用明显，最小容量阈值和一般状态评分明显升高（P<0.05，P<0.01），组内成分之间存在协同交互作用。结论：芍药苷+辛弗林+甘草酸组合对內脏高敏感大鼠结肠5-HT合成限速酶（TPH1）和相关受体（5-HT3R，5-HT4R）具有协同增效的调控作用，有效成分组合对5-HT通路的多靶点纠偏效应可能是四逆散改善大鼠内脏高敏感状态的重要机制。

Abstract

Objective: To explore the multi-target regulatory effect of paeoniflorin

synephrine

glycyrrhizin and their combination on 5-hydroxytryptamine(5-HT) signaling pathway[tryptophan hydroxylase 1(TPH1)/serotonin transporter(SERT)/5-HT3R/5-HT4R]

in order to explore the synergistic interaction between the ingredients and the action mechanism of the traditional Chinese medicine(TCM) prescription Sinisan on the visceral hypersensitivity rats. Method: The SD rats were randomly divided into normal group

model group

pinaverium bromide group

paeoniflorin group

synephrine group

glycyrrhizin group

paeoniflorin+glycyrrhizin group

paeoniflorin+synephrine group

glycyrrhizin + synephrine group

paeoniflorin + glycyrrhizin + synephrine group and Sinisan group. The irritable bowel(IBS) visceral hypersensitivity rat model was established by the comprehansive stress method. The intestinal sensitivity of the rats was evaluated by the minimum capacity threshold of abdominal withdrawal reflex (AWR). The protein and mRNA expressions of 5-HT

TPH1

SERT

5-HT3R

5-HT4R in colon were determined by enzyme linked immunosorbent assay(ELISA)

Western blot and Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). Result: Compared with the normal group

the minimum capacity threshold value of AWR significantly decreased in the model group (P<0.05)

with significant differences in protein and mRNA expressions of TPH1

5-HT3R

5-HT4R and SERT (P<0.05

P<0.01). Compared with the model group

the levels of 5-HT

TPH1

5-HT3R

and 5-HT4R were significantly improved

with an multi-target correction effect on TPH1

5-HT3R

5-HT4R mRNA expressions in the paeoniflorin + simvin + glycyrrhizin group (P<0.05

P<0.01). The value of minimum capacity threshold and the performance status significantly improved (P<0.05

P<0.01)

with an interaction between the components in the group. Conclusion: The combination of paeoniflorin

simvin and glycyrrhizin has a synergistic effect on the regulation of 5-HT synthase (TPH1) and related receptors (5-HT3R

5-HT4R) in the colon of visceral hypersensitivity rats. The multi-target rectifying effect of the active ingredient combination on the 5-HT pathway may be an important mechanism for Sinisan to alliviate the visceral hypersensitivity of rats.

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Related Institution

Institute of Basic Theory of Traditional Chinese Medicine， China Academy of Chinese Medical Sciences

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School of Chinese Materia Medica， Beijing University of Chinese Medicine

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Pharmaceutical Factory of Beijing Tongrentang Technology Development Co. Ltd.

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