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Measurement of rat gastric mucus secretion using ex-vivo gastric chamber system

使用离体胃室系统测量大鼠胃粘液分泌

基本信息

批准号：
03670377
负责人：
ISHIHARA Kazuhiko
金额：
$ 0.7万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

For the assessment of gastric mucus secreted from rat gastric mucosa into gastric lumen, we examined an ex-vivo gastric chamber system which had been developed by Takeuchi et al. To apply this system for the measurement of mucin secreted into the effluent solution, we had to develop a highly sensitive method to determine gastric mucin content. Finally we adopted an HPLC method to determine the content of hexosamine which constitutes the major carbohydarate component of mucin molecule. By using this method coupled with the removal treatment of low molecular weight hexosamine-containing materials including non-mucinous glycoproteins, it could be achieved to measure the time course changes of mucin secreted into the effluent solution. This method was applied to estimate the effect of carbachol administration on the mucus secretion in the rat gastric mucosa, and obtained a result that this drug to be an accelerator of mucus secretion into the gastric lumen.In the course of this study, we tried to obtain some monoclonal antibodies(MAbs) which are specific for the gastric mucin, and utilize them to the immunohistochemical method to determine the mucin content. We established several MAbs which are specific for the carbohydrate moieties of mucins present in the specific region of rat gastric mucosa. At present, we are trying to establish a method to assess the content of mucin originated from a specific site of gastric mucosa such as surface mucous cells, corpus gland mucous cells, antral gland mucous cells.

为了评估胃粘液从大鼠胃粘膜分泌到胃腔的情况，我们研究了Takeuchi等人开发的体外胃腔系统。为了应用这个系统来测量分泌到流出液中的粘蛋白，我们必须开发一种高灵敏度的方法来测定胃粘蛋白含量。最后，采用高效液相色谱法测定了粘蛋白分子的主要碳水酸盐成分氨基己糖的含量。将该方法与非粘性糖蛋白等低分子氨基己糖的去除处理相结合，可以实现对流出液中分泌的粘蛋白的时程变化的测量。应用该方法评价了卡巴胆碱对大鼠胃粘膜粘液分泌的影响，结果表明该药物是粘液分泌进入胃腔的促进剂。在本研究过程中，我们试图获得一些针对胃粘蛋白的单抗，并将其用于免疫组织化学方法测定粘蛋白含量。我们建立了几个针对大鼠胃粘膜特定区域粘蛋白碳水化合物部分的单抗。目前，我们正在试图建立一种方法来评估来自胃粘膜特定部位的粘蛋白的含量，如表面粘液细胞、体腺粘液细胞、胃窦腺粘液细胞。