Sini San ameliorates duodenal mucosal barrier injury and low-grade inflammation via the CRF pathway in a rat model of functional dyspepsia
Sini San ameliorates duodenal mucosal barrier injury and low-grade inflammation via the CRF pathway in a rat model of functional dyspepsia
复制标题
四逆散通过 CRF 通路改善功能性消化不良大鼠模型的十二指肠粘膜屏障损伤和低度炎症
DOI：
10.3892/ijmm.2019.4394
复制
发表时间：
2020-01-01
影响因子：
5.4
通讯作者：
Zhang, Shengsheng
中科院分区：
文献类型：
--
作者：
Zhu, Chunyang;Zhao, Luqing;Zhang, Shengsheng
肠-脑相互作用与十二指肠粘膜完整性受损和轻度炎症有关，已被证明是功能性消化不良（FD）的重要病理机制。四逆散是治疗FD的经典中药，但其作用机制尚不清楚。本研究的目的是评估SNS对FD患者十二指肠粘膜屏障损伤和轻度炎症的影响，并评估其对脑肠轴的潜在分子机制。采用碘乙酰胺法和尾部挤压法建立FD大鼠模型。免疫印迹法和/或免疫组化法检测促肾上腺皮质激素释放因子（CRF）、CRF受体1（CRF-R1）和CRF-R2的表达。此外，肥大细胞（MC）迁移进行了评估，通过免疫组化与抗类胰蛋白酶抗体，组胺浓度进行定量ELISA。逆转录-定量PCR检测类胰蛋白酶和蛋白酶激活受体2（PAR-2）mRNA表达水平，Western blot检测闭合蛋白1（ZO-1）、连接粘附分子1（JAM-1）、β-catenin和E-cadherin蛋白表达水平。结果表明，SNS治疗后，CRF在中枢神经系统和十二指肠的表达水平下调，SNS调节CRF-R1和CRF-R2的表达。此外，SNS抑制MC浸润和类胰蛋白酶/PAR-2途径在十二指肠的活性。SNS治疗可使FD大鼠ZO-1、JAM-1和β-catenin的表达水平恢复正常。这些结果表明，SNS对FD的治疗作用是通过恢复粘膜屏障完整性和抑制十二指肠的低度炎症来实现的，这至少部分是通过CRF信号通路介导的。
The gut-brain interaction is associated with impaired duodenal mucosal integrity and low-grade inflammation, which have been proven to be important pathological mechanisms of functional dyspepsia (FD). Sini San (SNS) is a classical Chinese medicine used to treat FD, but its underlying mechanisms are poorly understood. The aim of the present study was to evaluate the effects of SNS on duodenal mucosal barrier injury and low-grade inflammation with FD, and to assess its potential molecular mechanisms on the brain-gut axis. FD rats were established using the iodoacetamide and tail-squeezed methods. The expression of corticotropin-releasing factor (CRF), CRF receptor 1 (CRF-R1) and CRF-R2, were determined by western blot analysis and/or immunohistochemistry (IHC). In addition, mast cell (MC) migration was assessed by IHC with an anti-tryptase antibody, and histamine concentration was quantified using ELISA. The mRNA expression levels of tryptase and protease-activated receptor 2 (PAR-2) were quantified using reverse transcription-quantitative PCR, and the protein expression levels of zona occludens protein 1 (ZO-1), junctional adhesion molecule 1 (JAM-1), β-catenin and E-cadherin were determined via western blot analysis. It was demonstrated that the expression level of CRF was downregulated in the central nervous system and duodenum following SNS treatment, and that SNS modulated the expression of both CRF-R1 and CRF-R2. In addition, SNS suppressed MC infiltration and the activity of the tryptase/PAR-2 pathway in the duodenum. Furthermore, treatment with SNS restored the normal expression levels of ZO-1, JAM-1 and β-catenin in FD rats. These findings suggested that the therapeutic effects of SNS on FD were achieved by restoring mucosal barrier integrity and suppressing low-grade inflammation in the duodenum, which was at least partially mediated via the CRF signaling pathway.