"脾虚失运"的胃肠起搏细胞Cajal间质细胞机制研究

项目名称： "脾虚失运"的胃肠起搏细胞Cajal间质细胞机制研究

项目编号： No.30873328

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 电工技术

项目作者： 黄穗平

作者单位： 广州中医药大学

项目金额： 30万元

中文摘要： 在前期在体实验研究的基础上，初步提出"脾虚失运"的发生与胃肠起搏Cajal间质细胞（ICC）的结构和功能异常有关。本项目拟首先制作"脾虚失运"（即脾虚证胃肠动力障碍）大鼠模型，应用"以药测证"理论，观察健脾益气中药代表方四君子汤对其的影响，然后进一步从在体和体外实验两个方面探讨"脾虚失运"的ICC细胞机制；应用核素固体胃肠排空法观察脾虚大鼠胃肠排空；利用细胞微电极法观测胃肠平滑肌慢波活动；运用电子透镜、免疫组化法观测ICC的超微结构、数量、形态、ICC网络以及与胃肠神经系统、平滑肌细胞关系的变化；应用荧光定量PCR检测ICC标志基因c-kit的表达变化；应用细胞膜片箝、激光共聚焦显微镜观测ICC细胞电生理、细胞内游离钙离子浓度（［Ca2+］i）的动态变化。本项目将胃肠运动起搏理论和中医脾胃学说结合起来，以全新的角度阐明"脾虚失运"的发生机制，为中医脾胃学说现代研究开辟新的思路和方法。

中文关键词： 脾虚证；人参皂甙Rb1；ICC间质细胞；胃肠平滑肌细胞；香砂六君子汤

英文摘要： [Object] The research made gastric movement’ pacemaking cell and Interstitial Cell of Cajal (ICC) as the incision, probed into the pathological mechanism of gastric movement obstruction caused by spleen deficiency syndrome from the form and function of ICC, electricgastro- movement, the gastric emptying movement. It also can provide clinical and experimental proof for XiangSha Six Mild-Drug Decoction to treat spleen deficiency syndrome.To study the the effect of Ginsenoside Rb1 on the inward currents of ICCs and the mechanism of Rb1 effecting GI smooth muscle.To study the the effect of Ginsenoside Rb1 on the inward currents of ICCs and the mechanism of Rb1 effecting GI smooth muscle. [Methods]SD model rats suffered from spleen deficiency syndrome were used as objects, adopted gastric emptying detection, electron microscopic, immunity organization chemistry and RT-RCR techniques, etc. Before and after giving XiangSha Six Mild-Drug Decoction to the rats, observed the change of gastric motility. Form and distributing characteristics of ICC gastric pace-maker unit, gene-expressing variety of ICC cell marked substance c-kit. The small intestine of Balb/C mice (7–3 days old) of either sex was removed from 1cm below the pyloricring to the caecum, and luminal contents were washed out with Krebs–inger bicarbonate. Intestinal smooth muscle strips were isolated from male ICR mice (5 weeks old) and the spontaneous contraction was recorded by electrophysiolograph.Ion channel currents were recorded on freshly isolated single cells by using whole cell patch clamp technique. [Results] Dstails can be referred to report. [Conclusion] There was gastric emptying obstruction in patients suffered from the spleen deficiency syndrome of chronic gastritis.In patients suffered from the spleen deficiency syndrome of chronic gastritis, electrogastro- movement decreasing was the mechanism of electrical physiology of gastric emptying obstruction.In patients suffered from the spleen deficiency syndrome of chronic gastritis, the XiangSha Six Mild-Drug Decoction could improve dyspepsia symptoms and gastric emptying obstruction, its mechanism had relationship with its improving electrogastro- movement.In model rats suffered from spleen deficiency syndrome, there was abnormality in the form and function of ICC in gastric muscular layer, The XiangSha Six Mild-Drug Decoction could improve situation above. The experimental research results indicate that Ginsenoside Rb1 had no effect on the pacemaker current of cultured interstitial cells of Cajal fom murine small intestine. The experimental research results indicate that ginsenoside Rb1 exerted an inhibitory effect on the spontaneous contraction of intestinal smooth muscle in mouse, in the manner of decreasing the amplitude of the spontaneous contraction dose-dependently. The inhibitory effect of Rb1 is mediated by potentiating IKv and IKCa channel currents.

英文关键词： splenasthenic syndrome ；Ginsenoside Rb1; intestinal smooth muscle; intestinal smooth muscle cell；the XiangSha Six Mild-Drug Decoction