Heat shock protein 90 alpha and 14-3-3η in postmenopausal osteoporotic rats with varying levels of serum FSH

Abstract

Purpose

This study compared the severity of osteoporosis and screened differentially expressed proteins in postmenopausal osteoporotic rats with varying levels of serum follicle stimulating hormone (FSH).

Methods

Thirty-six Sprague Dawley female rats were divided into four groups: sham operation (sham) group, ovariectomy (OVX) group, FSH and ovariectomy (OVX + FSH) group, and Leuprorelin (LE) and ovariectomy group (OVX + LE). Body weight, serum estradiol, FSH, tartrate-resistant acid phosphatase, alkaline phosphatase, and bone mineral density were measured. We randomly selected six rats each from the OVX and OVX + FSH groups to detect differentially expressed proteins by data-independent acquisition, and we verified the results in the remaining six rats by enzyme-linked immunosorbent assay (ELISA).

Results

Nineteen proteins were upregulated and 36 proteins were downregulated in the OVX + FSH group. The expression of heat shock protein 90 alpha (Hsp90α) and 14-3-3η protein was significantly different between the OVX and OVX + FSH groups, and both were linearly correlated with bone trabecular area. Results were verified by ELISA and were found to be consistent with the results of data-independent acquisition.

Discussion

In rats with high serum FSH, expression of Hsp90α protein was increased and expression of 14-3-3η protein was decreased. Both changes in protein expression were strongly correlated with bone trabecular area.

摘要

目的：本研究在血清卵泡刺激素（FSH）不同水平的绝经后骨质疏松大鼠中, 比较了其骨质疏松症严重程度, 并筛选了差异表达的蛋白。

方法：将36只Sprague Dawley雌性大鼠分为四组：假手术（sham）组, 卵巢切除术（OVX）组, FSH和卵巢切除术（OVX + FSH）组以及亮丙瑞林（LE）和卵巢切除术组（OVX + LE） ）。测量体重, 血清雌二醇, FSH, 抗酒石酸酸性磷酸酶, 碱性磷酸酶和骨密度。我们从OVX和OVX + FSH组中随机选择六只大鼠, 通过数据独立采集来检测差异表达的蛋白质, 并通过酶联免疫吸附测定（ELISA）验证其余六只大鼠的结果。

结果：OVX + FSH组中有19种蛋白被上调, 而36种蛋白被下调。热休克蛋白90α（Hsp90α）和14-3-3η蛋白的表达在OVX组和OVX + FSH组之间有显著差异, 并且两者均与骨小梁面积呈线性相关。经ELISA方法进行验证后结果与数据独立采集的结果一致。

讨论：在高血清FSH的大鼠中, Hsp90α蛋白的表达增加, 而14-3-3η蛋白的表达减少。两种蛋白质的表达变化均与骨小梁面积密切相关。

Keywords:

• Postmenopausal osteoporosis
• rats
• follicle stimulating hormone
• heat shock protein 90 alpha
• 14-3-3η protein

Acknowledgements

The authors are grateful to all members of the Department of Gynecology, Nanjing Medical University, Affiliated Hangzhou Hospital (Hangzhou First People’s Hospital, Hangzhou Obstetrics & Gynecology Hospital). They thank Hangzhou Keruidi Technology Co., Ltd. and the animal experiment center of Zhejiang Chinese Medicine for providing technical support. They thank International Science Editing (http://www.internationalscienceediting.com) for editing this manuscript.

Potential conflict of interest

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work is financially supported by Zhejiang Health Committee (Major Projects Jointly Constructed by the Provincial and Ministry) [WKJ-ZJ-2010]; Health Leading Talents of Zhejiang Province [No.(2018)22]; Science and Technology Project of Zhejiang [2017C33203].