3种硬骨鱼类消化道肥大细胞组织的化学与免疫组化

Study on the mast cells in the digestive tract of three teleostean fishes by histochemical and immunohistochemical methods

类胰蛋白酶己被作为人类和某些哺乳动物组织中肥大细胞的标志。为检测鱼类肥大细胞胞浆中是否含有类胰蛋白酶,采用小鼠抗人类肥大细胞类胰蛋白酶单克隆抗体AA1通过ElivisionTM plus免疫组化染色技术,对尼罗罗非鱼、日本鳗鲡和欧洲鳗鲡消化道组织以及人胃癌组织石蜡切片中的肥大细胞进行染色,同时应用AB/SO和改良甲苯胺蓝法进行组织化学染色。结果表明,应用AB/SO和改良甲苯胺蓝染色都能较好显示尼罗罗非鱼肥大细胞,AB/SO染色法对日本鳗鲡和欧洲鳗鲡效果较差,改良甲苯胺蓝染色在上述两种鱼中未能检出肥大细胞;采用小鼠抗人类肥大细胞类胰蛋白酶单克隆抗体AA1通过ElivisionTM plus免疫组化染色技术,首次证实了尼罗罗非鱼肥大细胞胞浆中类胰蛋白酶的存在,但类胰蛋白酶阳性细胞数量很少,主要分布于尼罗罗非鱼消化道各段的黏膜上皮细胞基部和黏膜固有层,而日本鳗鲡与欧洲鳗鲡消化道均未检出类胰蛋白酶阳性细胞。说明不同鱼类肥大细胞胞浆颗粒中生物化学组分存在差别。人胃癌间质中可见多量类胰蛋白酶阳性细胞。

Tryptase has been considered as a cellular marker of mast cells in the tissues of human and some mammals. In order to detect whether or not fish mast cells contain tryptase in their cytoplasm, a murine monoclonal antibody （AA1）, raised against human mast cell tryptase , was used to stain the paraffin sections of the digestive tract tissue collected from Tilapia nilotica ,Anguilla japonica and AnguiUa anguilla and gastric cancer tissue（ as a positive control） from an adult man by ElivisionTMplus immunohistochemical techniques, Alcian blue Safranin O staining and the modified toluidine blue staining were used as histochemical staining at the same time. The results showed： Nile tilapia mast cells were displayed better by Alcian blue Safranin O staining and the modified toluidine blue staining, while the effects of A. japonica and A. anguiUa were not very good by Alcian blue Safranin O staining, and the mast cells could not be examined in A. japonica and A. anguilla by the modified toluidine blue staining. It was firstly demonstrated that Nile tilapia mast cells contain tryptase in their cytoplasm by Elivision^TMplus immunohistochemical techniques with a mufine monoclonal antibody AA1 raised against human mast cell tryptase. Small amount of the positive tryptase cells were found in Nile tilapia tissues and mostly lie in the base of mucosal epithelial cells and lamina propria in the sections of digestive tract. While the tryptase-positive mast cells had not been examined in A. japonica and A. anguilla. It showed that there was difference of biochemical element in cytoplasm granules among different fish mast cells. There were lots of tryptase-positive mast cells in the gastric carcinoma interstitium of human.