Zhu Feiye, Xu Shan, Zhang Yongsheng, Chen Fangming, Ji Jinjun, Xie Guanqun
Center of Analysis and Testing, Zhejiang Chinese Medical University, Hangzhou, China.
College of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou, China.
PLoS One. 2016 Aug 1;11(8):e0160398. doi: 10.1371/journal.pone.0160398. eCollection 2016.
Using an atropine-diphenoxylate-induced slow transit constipation (STC) model, this study explored the effects of the total glucosides of paeony (TGP) in the treatment of STC and the possible mechanisms.
A prospective experimental animal study.
The constipation model was set up in rats with an oral gavage of atropine-diphenoxylate and then treated with the TGP. The volume and moisture content of the faeces were observed and the intestinal kinetic power was evaluated. Meanwhile, the colorimetric method and enzyme linked immunosorbent assay (ELISA) were employed to determine the changes of nitric oxide (NO), nitric oxide synthase (NOS), vasoative intestinal peptide (VIP) and the P substance (SP) in the serum, respectively. The protein expressions of c-kit and stem cell factor (SCF) were assessed by immunohistochemical analysis and western blot, respectively, and the mRNA level of c-kit was measured by a reverse transcription polymerase chain reaction (RT-PCR).
The TGP attenuated STC responses in terms of an increase in the fecal volume and moisture content, an enhancement of intestinal transit rate and the reduction of NO, NOS and VIP in the serum. In addition, the c-kit, a labeling of interstitial cells of Cajal (ICC) increased at both protein and mRNA levels. SCF, which serves as a ligand of c-kit also increased at protein level.
The analysis of our data indicated that the TGP could obviously attenuate STC through improving the function of ICC and blocking the inhibitory neurotransmitters such as NO, NOS and VIP.
本研究采用阿托品 - 地芬诺酯诱导的慢传输型便秘(STC)模型,探讨白芍总苷(TGP)治疗STC的效果及可能机制。
一项前瞻性实验动物研究。
通过灌胃阿托品 - 地芬诺酯建立大鼠便秘模型,然后用TGP进行治疗。观察粪便体积和水分含量,并评估肠道动力。同时,分别采用比色法和酶联免疫吸附测定(ELISA)法测定血清中一氧化氮(NO)、一氧化氮合酶(NOS)、血管活性肠肽(VIP)和P物质(SP)的变化。分别通过免疫组织化学分析和蛋白质印迹法评估c-kit和干细胞因子(SCF)的蛋白表达,并通过逆转录聚合酶链反应(RT-PCR)测定c-kit的mRNA水平。
TGP可增加粪便体积和水分含量,提高肠道传输速率,降低血清中NO、NOS和VIP水平,从而减轻STC反应。此外,作为Cajal间质细胞(ICC)标志物的c-kit在蛋白和mRNA水平均增加。作为c-kit配体的SCF在蛋白水平也增加。
数据分析表明,TGP可通过改善ICC功能并阻断NO、NOS和VIP等抑制性神经递质来明显减轻STC。