Huntley J F, Newlands G F, Gibson S, Ferguson A, Miller H R
J Clin Pathol. 1985 Apr;38(4):375-84. doi: 10.1136/jcp.38.4.375.
Serine esterases were detected in the granules of mucosal mast cells from rat, mouse, sheep, and man. Successful demonstration of enzyme activity required brief fixation (6 h) of tissues in 4% paraformaldehyde. Staining with naphthol AS-D chloroacetate produced an intense red reaction product in intraepithelial mucosal mast cells (globule leucocytes) and mucosal mast cells within the lamina propria of the gastrointestinal tract. The mast cell identity of cells stained for esterase was confirmed by sequential staining with toluidine blue (pH 0.5). Furthermore, the numbers of cells detected after staining for esterases or with toluidine blue were highly correlated. Esterase activity within mucosal mast cells/globule leucocytes from all species was inhibited with the serine enzyme inhibitor phenylmethylsulphonyl fluoride. Further histochemical studies with the substrate, N-acetyl-DL-phenylalanine B-naphthyl ester, indicated that mucosal mast cells and globule leucocytes contain esterases which are chymotrypsin like in substrate specificity.
在大鼠、小鼠、绵羊和人的黏膜肥大细胞颗粒中检测到了丝氨酸酯酶。成功显示酶活性需要将组织在4%多聚甲醛中短暂固定(6小时)。用萘酚AS-D氯乙酸酯染色后,在上皮内黏膜肥大细胞(球形白细胞)和胃肠道固有层内的黏膜肥大细胞中产生了强烈的红色反应产物。通过用甲苯胺蓝(pH 0.5)进行连续染色,证实了酯酶染色细胞的肥大细胞身份。此外,酯酶染色或甲苯胺蓝染色后检测到的细胞数量高度相关。所有物种的黏膜肥大细胞/球形白细胞内的酯酶活性都被丝氨酸酶抑制剂苯甲基磺酰氟所抑制。用底物N-乙酰-DL-苯丙氨酸β-萘酯进行的进一步组织化学研究表明,黏膜肥大细胞和球形白细胞含有在底物特异性上类似于胰凝乳蛋白酶的酯酶。