Reed G B
Exp Mol Pathol. 1983 Apr;38(2):183-92. doi: 10.1016/0014-4800(83)90084-9.
Organ cultures prepared from 18- to 19-day fetal and 3- to 6-day-old newborn rat liver were maintained for 2 days in Trowell's T8 medium without insulin and supplemented with 0.1% albumin and 300 mg% glucose. The atmosphere for culture was 95% O2 and 5% CO2. Medium alone was used for control cultures, whereas insulin, hydrocortisone, or insulin plus hydrocortisone were used in experimental groups. Explant glycogen stores were maintained better in cultures grown in hormone-supplemented media than in control cultures. Fetal explants were found to have higher levels of glycogen than controls in the insulin or insulin plus hydrocortisone groups. Postnatal explants did not have higher levels of glycogen in the hormone-treated groups. Gestational age appeared to determine whether the liver explants reacted to hydrocortisone or insulin to maintain glycogen stores. Enzymatic assays, in vitro of glycogen synthetase and phosphorylase, indicated that the fetal liver response to insulin plus hydrocortisone by increasing the total and independent form of glycogen synthetase; but similar enzyme studies on postnatal rat liver did not show convincing differences as to an effect on synthetase. No definite in vitro temporal relationships could be identified. Late in gestation, the effect of hydrocortisone on glycogen synthesis is apparently dependent on the presence of insulin. Insulin appears to be required for glycogen storage in vitro in the cultures of postnatal rat liver.
从18至19天胎龄和3至6日龄新生大鼠肝脏制备的器官培养物,在不含胰岛素、补充有0.1%白蛋白和300mg%葡萄糖的特罗韦尔T8培养基中维持2天。培养环境为95%氧气和5%二氧化碳。单独的培养基用于对照培养,而实验组使用胰岛素、氢化可的松或胰岛素加氢化可的松。与对照培养相比,在补充激素的培养基中生长的培养物中外植体糖原储备维持得更好。在胰岛素或胰岛素加氢化可的松组中,发现胎儿外植体的糖原水平高于对照组。在激素处理组中,出生后外植体的糖原水平没有升高。胎龄似乎决定了肝脏外植体是否对氢化可的松或胰岛素产生反应以维持糖原储备。糖原合成酶和磷酸化酶的体外酶活性测定表明,胎儿肝脏通过增加糖原合成酶的总量和独立形式对胰岛素加氢化可的松产生反应;但对出生后大鼠肝脏进行的类似酶学研究未显示出对合成酶有令人信服的影响差异。无法确定明确的体外时间关系。在妊娠后期,氢化可的松对糖原合成的作用显然依赖于胰岛素的存在。胰岛素似乎是出生后大鼠肝脏培养物中体外糖原储存所必需的。