Wu Zhenyu, Lu Xiaofang, Zhang Shengsheng, Zhu Chunyang
Digestive Disease Center, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, China.
Evid Based Complement Alternat Med. 2020 Mar 31;2020:3575231. doi: 10.1155/2020/3575231. eCollection 2020.
The present study investigated the effect of Chinese medicine Sini-San (SNS) on visceral hypersensitivity in a rat model of functional dyspepsia (FD), and it explored related underlying mechanisms. The rat model of FD was developed by combining neonatal iodoacetamide (IA) treatment and adult tail-clamping. After SNS treatment, the behavior and electromyographic testing were performed to evaluate the visceromotor responses of rats to gastric distention. Immunofluorescence was used to detect the distribution of iNOS-positive cells in the spinal dorsal horn, while the real-time quantitative PCR and western blot were used for detection of the gene expression of c-fos, iNOS, and GABAb and protein levels of iNOS and GABAb in the spinal dorsal horn, respectively. The protein concentration of cGMP and PKG proteins in the spinal dorsal horn were quantified by enzyme-linked immunosorbent assay. In this study, SNS treatment significantly reduced the behavioral score and electromyographic response to graded intragastric distension pressure. The middle-dose of SNS treatment significantly reduced the distribution of iNOS-positive cells in the spinal dorsal horn of FD model rats. The gene expression of c-fos, iNOS, and GABAb and the protein contents of iNOS, GABAb, cGMP, and PKG in the spinal dorsal horn of FD model rats were restored to a normal level by middle-dose of SNS treatment. Our results suggest that Sini-San may alleviate the visceral hypersensitivity in FD model rats via regulation of the NO/cGMP/PKG pathway in the spinal dorsal horn.
本研究调查了中药四逆散（SNS）对功能性消化不良（FD）大鼠模型内脏高敏感性的影响，并探讨了相关潜在机制。FD大鼠模型通过新生期碘乙酰胺（IA）处理和成年期夹尾相结合的方法构建。SNS处理后，进行行为学和肌电图测试以评估大鼠对胃扩张的内脏运动反应。采用免疫荧光法检测脊髓背角中诱导型一氧化氮合酶（iNOS）阳性细胞的分布，同时分别采用实时定量PCR和蛋白质印迹法检测脊髓背角中c-fos、iNOS和γ-氨基丁酸B型受体（GABAb）的基因表达以及iNOS和GABAb的蛋白水平。采用酶联免疫吸附测定法对脊髓背角中cGMP和蛋白激酶G（PKG）蛋白的浓度进行定量。在本研究中，SNS处理显著降低了对分级胃内扩张压力的行为评分和肌电图反应。SNS中剂量处理显著减少了FD模型大鼠脊髓背角中iNOS阳性细胞的分布。SNS中剂量处理使FD模型大鼠脊髓背角中c-fos、iNOS和GABAb的基因表达以及iNOS、GABAb、cGMP和PKG的蛋白含量恢复至正常水平。我们的结果表明，四逆散可能通过调节脊髓背角中的一氧化氮/环磷酸鸟苷/蛋白激酶G通路来减轻FD模型大鼠的内脏高敏感性。
