尼罗罗非鱼消化道肥大细胞的组化性质

实验采用改良甲苯胺蓝(MTB)、阿利新蓝-沙黄(AB/SO)、甲基绿-派洛宁(MG-P)、天青Ⅱ-伊红-瑞氏混合液和硫堇5种组化染色法,对尼罗罗非鱼(Nile tilapia)消化道组织中的肥大细胞(Mast cell,MC)组化性质进行研究。尼罗罗非鱼的食管、胃及小肠壁内均显示有肥大细胞,在食管和胃的切片标本上肥大细胞主要分布在黏膜固有层和胃腺体之间。在肠道中的肥大细胞主要分布在黏膜固有层和肠上皮下方,少量肥大细胞存在于黏膜下层结缔组织中。细胞呈圆形、椭圆形,也有长梭形的。而且肥大细胞有沿血管分布的特点。5种组化染色结果表明:AB/SO、MTB和MG-P显示的MC效果较好,尤其AB/SO染色效果最好,肥大细胞轮廓清楚,胞质颗粒较清晰;尼罗罗非鱼肥大细胞胞浆颗粒都呈红色,即肥大细胞胞浆主要含肝素,不含组胺。天青Ⅱ-伊红-瑞氏混合液染色效果也很好,但被染的肥大细胞较少;80%乙醇硫堇染色,在尼罗罗非鱼消化道各段组织中均未能鉴定出肥大细胞。尼罗罗非鱼消化道肥大细胞大多分布于浅层的黏膜或血管、腺体周围的结缔组织等易表露于环境抗原的位点。罗非鱼消化道黏膜层结缔组织中的肥大细胞与大多数脊椎动物的肥大细胞一样,具有沿血管分布的特性,说明硬骨鱼的肥大细胞如哺乳动物肥大细胞一样与血管有着密切的关系。  

STUDY ON THE HISTOCHEMICAL PROPERTY OF THE MAST CELLS IN THE DIGESTIVE TRACT OF NILE TILAPIA

Five healthy nile tilapia(average weight 500g) were bought.After executing,the esophagus,stomachus(stomachus fundus,stomachus pylorus and stomachus cardiacus),small intestine(foregut,midgut and hindgut) were respectively fixed in the Carnoy solution(60% dehydrate alcohol,30% chloroform and 10% glacial acetic acid),con-tinuously embedded and sliced with 6μm.Toluidine blue staining(MTB),Alcian blue Safranin O staining(AB/SO),methyl green-Pyronine staining(MG-P),Azure Ⅱ-Eosin-Wright mixture staining and Thionine staining were used to investigate the histochemical property of the mast cells in the digestive tract of tilapia. The mast cells were showed in the esophagus,stomachus and small intestinal wall of nile tilapia.The mast cells in the slides of esophagus and stomachus mostly distributed between lamina propria and stomachus gland.The mast cells in intestinal tract mainly distributed in lamina propria and under intestine epithelium,while less in connective tissue of submucosa.They were circular,ellipse and spindle,and intended to be around blood vessels.The results of five histo-chemical stainings were as follows: The MTB,AB/SO and MG-P stainings could show the mast cells better,especially the AB/SO was the best among them.The mast cells were distinctly outlined,the cytoplasmic particle of which was clear.The similarity of the research on teleostean mast cells from home and abroad was that the mast cells of nile tilapia was similar to mucosal mast cell of mammal.The effect of MG-P staining was good too,in which the cytoplasmic par-ticle of mast cells showed red,which meant the cytoplasmic contained mainly heparin not histamine.It confirmed fur-ther that the histochemical property of the mast cells of nile tilapia was similar to mucosal mast cell of mammal.The effect of Azure Ⅱ-Eosin-Wright mixture staining was better,but the mast cells dyed was few.Whereas,the mast cells in the sections of digestive tract of nile tilapia could not been showed by 80% Alcohol-Thionine staining.The reason might be different stationary liquid or the difference in histochemical property of mast cells of nile tilapia,carp and white fish.The observation showed that the mast cells in the digestive tract of nile tilapia mostly distributed in the points which easily revealed from environment antigen such as shallow mucosa or blood vessels,the connective tissue around gland and so on.The mast cells had similar distribution characteristics in different organs of other animals.From the above,the distribution characteristics of creatural mast cells were closely bound up with the function of cell,which maybe provide handy positional foundation for the exertion of autoimmune function in a sense.The experiment also found that the location of mast cells in connective tissue of mucosa in digestive tract of nile tilapia were similar with most of the vertebrate,of which the location was intended to be around blood vessels.It meant that teleostean mast cells were closely related to blood vessels as mammals were. It was reported that the growth of vascular endothelial cells and constitute of blood circulation were affected by histamine and heparin released from mast cells.The vascular endothelial leukocyte adhesion molecule 1,which could watch blood stream by attracting lymphocyte adhere to the inner wall of blood vessels,could be induced by Tumor Ne-crosis Factor-α(TNF-α) released from mast cells in human skin.It was obviously that the mast cells around blood ves-sels played an important role in reaction of blood vessels.