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小鼠胎盘细胞分泌可溶性瘦素受体(sOB-R):环磷酸腺苷(cAMP)抑制sOB-R的产生。

Mouse placental cells secrete soluble leptin receptor (sOB-R): cAMP inhibits sOB-R production.

作者信息

Yamaguchi M, Murakami T, Yasui Y, Otani S, Kawai M, Kishi K, Kurachi H, Shima K, Aono T, Murata Y

机构信息

Department of Obstetrics and Gynecology, Osaka University Medical School, Suita, 565-0871, Japan.

出版信息

Biochem Biophys Res Commun. 1998 Nov 18;252(2):363-7. doi: 10.1006/bbrc.1998.9636.

Abstract

The aims of this study were to identify whether mouse placenta secretes soluble OB-R (sOB-R) and to find the regulating factor of OB-R expression. Total RNAs were extracted from placenta and decidua, and OB-R expression was assessed by Northern blot analysis. Decidua did not express OB-R mRNA. However, OB-R mRNA expression was detectable in the placenta on day 13 of pregnancy, and then it increased and reached a peak on day 17 of pregnancy. Mouse placental cells from day 12 of pregnancy were cultured and OB-R gene expression was assessed by Northern blot analysis. OB-R mRNA expression was detectable from the second day of culture and reached a peak on the third day of culture. To determine whether placental cells release sOB-R, supernatant of cultured placental cells was subjected to Western blot analysis. sOB-R was detected in the medium by the second day of culture and sOB-R release increased up to the fourth day of culture. Addition of leptin to the medium did not affect expression of OB-R mRNA. However, 8-bromo cAMP inhibited both steady-state levels of OB-R mRNA and the amount of sOB-R protein in the medium in a dose- and time-dependent manner. These results suggest that trophoblast cells differentiate, express, and release sOB-R both in vivo and in vitro and that cAMP is one of several potent regulators of sOB-R secretion by the mouse placenta.

摘要

本研究的目的是确定小鼠胎盘是否分泌可溶性OB-R(sOB-R),并寻找OB-R表达的调节因子。从胎盘和蜕膜中提取总RNA,通过Northern印迹分析评估OB-R的表达。蜕膜不表达OB-R mRNA。然而,在妊娠第13天的胎盘中可检测到OB-R mRNA表达,然后其表达增加并在妊娠第17天达到峰值。培养妊娠第12天的小鼠胎盘细胞,通过Northern印迹分析评估OB-R基因表达。培养第二天即可检测到OB-R mRNA表达,并在培养第三天达到峰值。为了确定胎盘细胞是否释放sOB-R,对培养的胎盘细胞上清液进行Western印迹分析。培养第二天在培养基中检测到sOB-R,并且sOB-R释放量在培养第四天之前持续增加。向培养基中添加瘦素不影响OB-R mRNA的表达。然而,8-溴环磷酸腺苷(8-bromo cAMP)以剂量和时间依赖性方式抑制OB-R mRNA的稳态水平以及培养基中sOB-R蛋白的量。这些结果表明,滋养层细胞在体内和体外均可分化、表达并释放sOB-R,并且cAMP是小鼠胎盘sOB-R分泌的几种有效调节因子之一。

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