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大鼠孕期胎盘催乳素-II对胰岛素分泌及胰岛β细胞增殖的体外研究

In vitro studies of the stimulation of insulin secretion and B-cell proliferation by rat placental lactogen-II during pregnancy in rats.

作者信息

Kawai M, Kishi K

机构信息

New Drug Research Laboratories, Shionogi & Co., Ltd, Osaka, Japan.

出版信息

J Reprod Fertil. 1997 Jan;109(1):145-52. doi: 10.1530/jrf.0.1090145.

Abstract

The present study investigated the effect of rat placental lactogen-II (rPL-II) on insulin secretion and B-cell proliferation of the maternal islets during the last half of pregnancy in rats using a homologous system. Pancreatic islets were isolated from nonpregnant rats and rats at day 13 of pregnancy and cultured for 8 days in medium containing trophoblast culture medium or purified hormones. The medium was changed daily and insulin concentrations were determined by measuring immunoreactivity. The number of proliferating B cells were determined by double staining for both insulin and 5'-bromo-2'-deoxyuridine (BrdU) incorporated into replicating DNA during the last 24 h of incubation. rPL-II-enriched medium, in which trophoblasts of placentae from rats at day 13 of pregnancy were incubated for 8 days, was added to the islet culture system. Insulin concentration in the medium of non-pregnant rat islets was significantly increased and doubled on incubation in 100% trophoblast culture medium. Addition of purified rPL-II to the culture medium of pregnant rat islets stimulated insulin secretion at the concentrations of 50-500 ng ml-1 in a dose-dependent manner. The stimulatory effect of rPL-II on insulin secretion was found to be more than double that with rat prolactin (rPRL) and rat growth hormone (rGH) at 100 ng ml-1. For determination of B-cell proliferation, non-pregnant rat islets were incubated with rPL-II, rPRL or rGH at 1000 ng ml-1 for 8 days and then 10 mmol BrdU l-1 was added during the last 24 h of incubation. rPL-II (5.30 +/- 0.36%), rPRL (3.79 +/- 0.34%) and rGH (2.87 +/- 0.29%) significantly increased the rate of incorporation of BrdU into the B cells compared with that of the control (1.34 +/- 0.18%). These results indicate that rPL-II directly regulates insulin secretion and B-cell proliferation of maternal islets during the last half of pregnancy in rats.

摘要

本研究使用同源系统,调查了大鼠胎盘催乳素-II(rPL-II)对大鼠妊娠后半期母体胰岛胰岛素分泌和B细胞增殖的影响。从未孕大鼠和妊娠第13天的大鼠分离胰岛,并在含有滋养层培养基或纯化激素的培养基中培养8天。每天更换培养基,并通过测量免疫反应性来测定胰岛素浓度。通过对胰岛素和在孵育最后24小时掺入复制DNA中的5'-溴-2'-脱氧尿苷(BrdU)进行双重染色,来确定增殖B细胞的数量。将富含rPL-II的培养基(其中妊娠第13天大鼠胎盘的滋养层孵育8天)添加到胰岛培养系统中。未孕大鼠胰岛培养基中的胰岛素浓度在100%滋养层培养基中孵育时显著增加并翻倍。向妊娠大鼠胰岛培养基中添加纯化的rPL-II,在50 - 500 ng/ml的浓度下以剂量依赖方式刺激胰岛素分泌。发现rPL-II对胰岛素分泌的刺激作用在100 ng/ml时比大鼠催乳素(rPRL)和大鼠生长激素(rGH)的刺激作用高出一倍多。为了测定B细胞增殖,将未孕大鼠胰岛与1000 ng/ml的rPL-II、rPRL或rGH孵育8天,然后在孵育的最后24小时添加10 mmol BrdU/l。与对照组(1.34±0.18%)相比,rPL-II(5.30±0.36%)、rPRL(3.79±0.34%)和rGH(2.87±0.29%)显著增加了BrdU掺入B细胞的速率。这些结果表明,rPL-II在大鼠妊娠后半期直接调节母体胰岛的胰岛素分泌和B细胞增殖。

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