Jeffrey J J, Roswit W T, Ehlich L S
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
J Cell Physiol. 1990 May;143(2):396-403. doi: 10.1002/jcp.1041430226.
An enzyme-linked immunosorbent assay (ELISA) has been developed for rat collagenase. The assay is capable of measuring the enzyme from a variety of rat cell sources at concentrations of 10-50 ng/ml, approximately 500-1,000-fold more sensitive than radiolabelled collagen fibril assay systems. The assay is specific to collagenase from the rat: enzymes from human, tadpole, mouse, and bacterial sources failed to cross-react significantly with rat enzyme. The assay is reproducible and accurate, and is capable of detecting enzyme in the presence of serum or tissue inhibitors. Using the ELISA, we have examined the effect of a variety of hormones on the production of collagenase by rat myometrial smooth muscle cells in culture. Of all the reproductive hormones examined, only progesterone and its synthetic derivative medroxyprogesterone acetate were capable of inhibiting the production of the enzyme by these cells. The maximally effective concentration of progesterone was 1 x 10(-6)M, and that of medroxyprogesterone acetate was 1 x 10(-7)M. The effect of the steroid was selective: no effect on cell proliferation or on general protein synthesis was observed. In addition to the progestational steroids, the glucocorticoids were also capable of inhibiting the production of collagenase by the cells at similar nominal concentrations. However, the myometrial cells were found actively to metabolize progesterone but not hydrocortisone in culture. Thus, the effective inhibitory concentration of progesterone was approximately ten-fold lower than that of hydrocortisone. The results of this study support the concept that progesterone plays a major role in preventing the production of collagenase in the rat uterus.
已开发出一种用于大鼠胶原酶的酶联免疫吸附测定法（ELISA）。该测定法能够检测来自多种大鼠细胞来源的酶，检测浓度范围为10 - 50 ng/ml，比放射性标记的胶原纤维测定系统灵敏约500 - 1000倍。该测定法对大鼠胶原酶具有特异性：来自人类、蝌蚪、小鼠和细菌来源的酶与大鼠酶无明显交叉反应。该测定法具有可重复性和准确性，并且能够在存在血清或组织抑制剂的情况下检测酶。使用ELISA，我们研究了多种激素对培养的大鼠子宫肌层平滑肌细胞胶原酶产生的影响。在所检测的所有生殖激素中，只有孕酮及其合成衍生物醋酸甲羟孕酮能够抑制这些细胞产生该酶。孕酮的最大有效浓度为1×10⁻⁶M，醋酸甲羟孕酮的最大有效浓度为1×10⁻⁷M。类固醇的作用具有选择性：未观察到对细胞增殖或一般蛋白质合成的影响。除了孕激素类固醇外，糖皮质激素在相似的标称浓度下也能够抑制细胞产生胶原酶。然而，发现子宫肌层细胞在培养中能积极代谢孕酮但不能代谢氢化可的松。因此，孕酮的有效抑制浓度比氢化可的松低约10倍。本研究结果支持孕酮在阻止大鼠子宫中胶原酶产生方面起主要作用这一概念。
