Gutschmidt S, Menge H, Riecken E O
Lab Invest. 1982 May;46(5):476-80.
The in situ kinetic constants of neutral alpha- glucosidase were determined in the atrophic jejunal mucosa of self-emptying blind loops and in corresponding jejunal segments of control rats, using a quantitative histochemical technique. The apparent maximum velosity (Vmax) and km values were calculated from absorbance measurements of an azo dye-deposit formed in the brush border membrane at the villus base and at the transition zone between the middle and upper one-third of the villus. In the controls a significant increase of apparent Vmax and substrate affinity (decrease of apparent Km) was obtained, corresponding to enterocyte maturation along the villi. The mucosal atrophy in self-emptying blind loops was accompanied by a significant decrease of apparent Vmax of neutral alpha-glucosidase at both representative villus sites, but the substrate affinity was increased at both sites. The results indicate that alpha-glucosidase activity was reduced when measured at the cellular level of two representative villus sites in the atrophic jejunal mucosa, possibly due to the absence of substrate-effected enzyme induction. The decrease on hydrolytic enzyme activity was not matched by a compensatory increase in substrate affinity.
采用定量组织化学技术,测定了自行排空盲袢萎缩空肠黏膜以及对照大鼠相应空肠段中中性α-葡萄糖苷酶的原位动力学常数。通过对在绒毛基部以及绒毛中上部与上部三分之一交界处的刷状缘膜中形成的偶氮染料沉积物的吸光度测量,计算出表观最大速度(Vmax)和Km值。在对照中,随着沿绒毛的肠上皮细胞成熟,表观Vmax和底物亲和力显著增加(表观Km降低)。自行排空盲袢中的黏膜萎缩伴随着在两个代表性绒毛部位中性α-葡萄糖苷酶的表观Vmax显著降低,但两个部位的底物亲和力均增加。结果表明,在萎缩空肠黏膜的两个代表性绒毛部位的细胞水平上进行测量时,α-葡萄糖苷酶活性降低,这可能是由于缺乏底物效应的酶诱导所致。水解酶活性的降低并未伴随着底物亲和力的代偿性增加。