Triantafyllou Asterios, Fletcher David, Scott John
Department of Oral Pathology, The University of Liverpool School of Dentistry, Pembroke Place, Liverpool L3 5PS, UK.
Histochem J. 2002;34(11-12):529-33. doi: 10.1023/a:1026037801958.
Catalytic activities of lingual lipase were investigated by enzyme histochemistry in post-mortem tongues from male rats. Sections of fresh-frozen or formalin-calcium fixed tissue were incubated with naphthol-AS-nonanoate and alpha-naphthyl acetate substrate mixtures. The effects of pH level, sodium taurocholate activator and E600 inhibitor were also examined. The use of cryostat sections of tissues fixed in formalin-calcium and of nonanoate substrate within the range of pH 4.4-6.4, were optimal for localizing maximum reaction product, captured by Fast Blue BB, in acini and demilunes of the posterior deep and superficial lingual glands respectively. The reaction product corresponded with the distribution of secretory granules and failed to develop when taurocholate was omitted from the incubation medium. Similarly localized E600-resistant reaction product occurred with the acetate substrate and hexazotized New Fuchsin at pH 7.4, in the absence of taurocholate. Lipase and conventional esterase activities appear to be superimposed in posterior lingual glands of rat. The ability of their acini and demilunes to hydrolyse nonanoate substrate at an acidic pH optimum, when activated by sodium taurocholate, seems attributable to lipase destined for secretion into saliva--hence convenient for routine histochemical identification of the enzyme.
通过酶组织化学方法，对雄性大鼠死后的舌头进行研究，以探讨舌脂肪酶的催化活性。将新鲜冷冻或经福尔马林 - 钙固定的组织切片与萘酚 - AS - 壬酸酯和α - 萘乙酸底物混合物一起孵育。还研究了pH值、牛磺胆酸钠激活剂和E600抑制剂的影响。使用经福尔马林 - 钙固定的组织的低温恒温器切片以及在pH 4.4 - 6.4范围内的壬酸酯底物，最适合将通过固蓝BB捕获的最大反应产物分别定位在舌后深部和浅表腺泡和半月形腺泡中。反应产物与分泌颗粒的分布相对应，并且当孵育培养基中省略牛磺胆酸钠时反应不发生。同样，在不存在牛磺胆酸钠的情况下，在pH 7.4时，乙酸底物和六偶氮新福林会出现类似定位的E600抗性反应产物。脂肪酶和传统酯酶活性似乎在大鼠舌后腺中重叠。当被牛磺胆酸钠激活时，它们的腺泡和半月形腺泡在酸性pH最佳条件下水解壬酸酯底物的能力似乎归因于注定要分泌到唾液中的脂肪酶，因此便于对该酶进行常规组织化学鉴定。
