卵泡刺激素受体在软骨细胞中的表达及卵泡刺激素对软骨细胞的影响。

Expression of FSHR in chondrocytes and the effect of FSH on chondrocytes.

作者信息

Kong Dehuan, Guan Qingbo, Li Guandong, Xin Wei, Qi Xiaoyi, Guo Yanjing, Zhao Jiajun, Xu Jin, Sun Shui, Gao Ling

机构信息

Department of Endocrinology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong province, China; Shandong Provincial Key Laboratory of Endocrinology and Lipid Metabolism, Jinan, Shandong province, China; Institute of Endocrinology and Metabolism, Shandong Academy of Clinical Medicine, Jinan, Shandong province, China; Department of Geriatrics, Taian City Central Hospital, Taian, Shandong province, China.

出版信息

Biochem Biophys Res Commun. 2018 Jan 1;495(1):587-593. doi: 10.1016/j.bbrc.2017.11.053. Epub 2017 Nov 10.

DOI:10.1016/j.bbrc.2017.11.053

PMID:29133260

Abstract

OBJECTIVE

Chondrocytes express many kinds of hormone receptors. The function of Follicle stimulating hormone (FSH) in the ovary is mediated by FSH receptor (FSHR). FSH receptor (FSHR) is found in many non-ovarian tissues, however it has been unclear if chondrocytes express FSHR. The purpose of this study is to determine it.

METHODS

Mouse primary chondrocytes and human articular cartilage tissues were examined. The expression and sequence of FSHR mRNA by reverse transcription polymerase chain reaction (RT-PCR) and sequenced, respectively, and its protein expression was tested using western blotting and location was observed under immunofluorescence microscopy. Ovarian tissue was as a positive control. After FSH stimulated mouse chondrocytes, intracellular cAMP levels were assessed by ELISA, and gene expression relative to Mouse WNT Signaling Pathway was tested by RT Profiler PCR Arrays.

RESULTS

FSHR was detected at the transcriptional level and confirmed to have the same sequence as that of ovary-derived mRNA of FSHR. FSHR proteins presented at the same line as the positive proteins of ovary, in mouse chondrocytes and human cartilage tissue, respectively. FSHR proteins were located at the cell membrane. Intracellular cAMP contents were significantly elevated up to 7-fold in mouse chondrocytes by forskolin (100 mM) (P < 0.001); however, different doses of FSH did not change the cAMP contents in mouse primary chondrocytes. RT Profiler PCR Arrays demonstrated that FSH could cause changes in gene expression among the 84 preordained genes, such as Fosl1, Rhou, and Dkk1, in mouse chondrocytes relative to the control.

CONCLUSION

Mouse chondrocytes and human articular cartilage express functional FSHR. Moreover, FSH can act on chondrocytes and cause genetic changes.

摘要

目的

软骨细胞表达多种激素受体。促卵泡激素（FSH）在卵巢中的功能由促卵泡激素受体（FSHR）介导。FSH受体（FSHR）存在于许多非卵巢组织中，但软骨细胞是否表达FSHR尚不清楚。本研究旨在确定这一点。

方法

对小鼠原代软骨细胞和人关节软骨组织进行检测。分别通过逆转录聚合酶链反应（RT-PCR）检测FSHR mRNA的表达和序列，并进行测序，利用蛋白质免疫印迹法检测其蛋白表达，并在免疫荧光显微镜下观察其定位。卵巢组织作为阳性对照。FSH刺激小鼠软骨细胞后，通过酶联免疫吸附测定（ELISA）评估细胞内cAMP水平，并通过RT Profiler PCR Array检测与小鼠WNT信号通路相关的基因表达。

结果

在转录水平检测到FSHR，并证实其序列与卵巢来源的FSHR mRNA相同。FSHR蛋白在小鼠软骨细胞和人软骨组织中分别与卵巢阳性蛋白位于同一位置。FSHR蛋白位于细胞膜上。福斯高林（100 mM）可使小鼠软骨细胞内cAMP含量显著升高至7倍（P < 0.001）；然而，不同剂量的FSH并未改变小鼠原代软骨细胞中的cAMP含量。RT Profiler PCR Array表明，相对于对照组，FSH可导致小鼠软骨细胞中84个预定基因（如Fosl1、Rhou和Dkk1）的基因表达发生变化。