抗人卵泡刺激素受体多克隆抗体的制备及其对实验大鼠骨质疏松的抑制作用

doi:10.13523/j.cb.20170602

中国生物工程杂志

2017, Vol. 37

Issue (6): 9-16 DOI: 10.13523/j.cb.20170602

研究报告

抗人卵泡刺激素受体多克隆抗体的制备及其对实验大鼠骨质疏松的抑制作用

秦瑞坪, 李玲霞, 马晓玲, 席欧彦, 赵婷, 邱玲玲, 李江伟

新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室乌鲁木齐 830046

Inhibition of Osteoporosis in Ovariectomized Rats Using Follicle-stimulating Hormone Receptor Specific Polyclonal Antibody

QIN Rui-ping, LI Ling-xia, MA Xiao-ling, XI Ou-yan, ZHAO Ting, QIU Ling-ling, LI Jiang-wei

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China

全文: PDF(1388 KB) HTML

摘要： 近年来，卵泡刺激素（follicle-stimulating hormone，FSH）在骨质疏松中的作用得到了证实。卵泡刺激素受体（FSH receptor，FSHR）在成熟的人破骨细胞及破骨细胞前体即单核细胞表面表达，成为阻断FSH 作用的潜在靶点。制备了高滴度的兔抗人FSHR 多克隆抗体，采用双侧去卵巢手术方法建立SD （Sprague-Dawley）大鼠骨质疏松症动物模型，观察抗FSHR抗体对大鼠实验骨质疏松症的治疗效果。结果显示，卵巢摘除（ovariectomized，OVX）大鼠经抗FSHR多抗和亮丙瑞林（leuprorelin，LE）治疗两周后，与PBS对照组相比，血清FSH和黄体酮（luteinizing hormone，LH）水平显著降低（P<0.05），而雌二醇（estrogen，E2）水平有一定升高，但结果不显著（P>0.05）。另外，骨组织化学染色显示多抗和LE治疗组大鼠骨小梁数目增加，断裂现象较少。这些结果初步表明，采用抗FSHR抗体对SD 大鼠骨质疏松症具有一定的治疗作用。

关键词： 骨质疏松症; SD大鼠; 亮丙瑞林; 抗FSHR多克隆抗体; 卵泡刺激素受体

Abstract: There are strong evidences to support the idea that follicle-stimulating hormone (FSH) plays an important role in regulation of bone mass. Owing to its limited expression to gonadal tissues and osteoclasts and its precursor monocytes, FSH receptor may be a good target for FSH blocking therapy. High titers of polyclonal rabbit anti-human FSHR antibody (pAb_FSHR) was raised and prepared. The bilateral ovariectomized (OVX) Sprague-Dawley (SD) rats were used as the osteoporosis model for evaluating the effects of pAb_FSHR and FSH inhibitor leuprorelin(LE) on the bone loss. The results showed after treatment with pAb_FSHR and LE separately, the FSH and luteinizing hormone(LH) levels in OVX groups decreased significantly (P<0.05) compared with PBS treated OVX rats. No significant differences of estrogen (E2) levels were detected between treatment and PBS injected OVX rats (P>0.05). In addition, the number of trabecular bone increased after pAb_FSHR treatment in OVX groups in the shin bone histologic analysis. The results suggested targeting against FSHR may work on the treatment of osteoporosis.

Key words: Osteoporosis anti-FSHR polyclonal antibody Leuprorelin SD rats Follicle-stimulating hormone receptor

收稿日期: 2016-12-27 出版日期: 2017-06-25

ZTFLH:

R392.1

基金资助: 国家自然科学基金资助项目（81260333）

通讯作者: 李江伟 E-mail: jwli67@sina.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	赵婷
	秦瑞坪
	马晓玲
	席欧彦
	邱玲玲
	李玲霞
	李江伟

引用本文:

秦瑞坪, 李玲霞, 马晓玲, 席欧彦, 赵婷, 邱玲玲, 李江伟. 抗人卵泡刺激素受体多克隆抗体的制备及其对实验大鼠骨质疏松的抑制作用[J]. 中国生物工程杂志, 2017, 37(6): 9-16.

QIN Rui-ping, LI Ling-xia, MA Xiao-ling, XI Ou-yan, ZHAO Ting, QIU Ling-ling, LI Jiang-wei. Inhibition of Osteoporosis in Ovariectomized Rats Using Follicle-stimulating Hormone Receptor Specific Polyclonal Antibody. China Biotechnology, 2017, 37(6): 9-16.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170602 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I6/9

[1] Kim J H, Kim E Y, Lee B, et al. The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells. International Journal of Molecular Medicine, 2016,37(3):649-658.
[2] 潘德, 郝秋芳, 王玮,等. 绝经老年妇女骨质疏松调查及影响因素分析. 中国老年学杂志, 1994,14(6):352-354. Pan D, Hao Q F, Wang W,et al. Older women menopause osteoporosis research and influence factors analysis. Chinese Journal of Gerontology,1994,14(6):352-354.
[3] Zhu L L, Blair H, Cao J,et al. Blocking antibody to the β-subunit of FSH prevents bone loss by inhibiting bone resorption and stimulating bone synthesis. Proceedings of the National Academy of Sciences, 2012, 109(36):14574-14579.
[4] Cannon J G, Cortez-Cooper M, Meaders E, et al. Follicle-stimulating hormone, interleukin-1, and bone density in adult women. American Journal of Physiology-Regulatory,Integrative and Comparative Physiology, 2010, 298(3):R790-R798.
[5] Wang J, Zhang W, Yu C, et al. Follicle-stimulating hormone increases the risk of postmenopausal osteoporosis by stimulating osteoclast differentiation. PloS One, 2015, 10(8):e0134986.
[6] Liu S, Cheng Y, Xu W, et al. Protective effects of follicle-stimulating hormone inhibitor on alveolar bone loss resulting from experimental periapical lesions in ovariectomized rats. Journal of endodontics, 2010, 36(4):658-663.
[7] Sun L, Peng Y, Sharrow A C, et al. FSH directly regulates bone mass. Cell, 2006, 125(2):247-260.
[8] Sun L, Zhang Z, Zhu L L, et al. Further evidence for direct pro-resorptive actions of FSH.Biochemical and Biophysical Research Communications, 2010, 394(1):6-11.
[9] 夏雪琴, 木亚沙尔·买买提拉洪, 翟田甜,等. 抗卵泡刺激素受体纳米抗体的制备及鉴定. 细胞与分子免疫学杂志, 2013, 29(8):829-833. Xia X Q, Mu Y S, Zhai T T,et al. Preparation and identification of anti-follicle-stimulating hormone receptor nanobodies. Chin J Cell Mol Immunol,2013,29(8):829-833.
[10] 卢勇, 孟庆才, 方锐,等. SD大鼠骨性关节炎合并骨质疏松症模型的建立. 中国组织工程研究与临床康复, 2009, 13(46):9092-9096. Lu Y, Meng Q C, Fang R,et al. Establishment of SD rat models of osteoarthritis and osteoporosis. Journal of Clinical Rehabilitative Tissue Engineering Research,2009,13(46):9092-9096.
[11] 田茂友, 李洪洋.去卵巢大鼠骨质疏松模型研究. 现代预防医学, 2007, 34(12):2239-2241. Tian M Y, Li H Y. Experimental study on the model of osteoporosis in ovariectomized rats[J].Modern Preventive Medicine,2007,34(12):2239-2241.
[12] 李旭鸿, 侯曼, 仰红慧. 骨密度的测定方法及影响因素.天津体育学院学报, 2005, 20(3):62-65. Li H X, Hou M, Yang H H. The measurement methods and influence factors of bone density. Journal of TjIPE,2005,20(3):62-65.
[13] Xiaodong L, Ominsky M S, Warmington K S, et al. Sclerostin antibody treatment increases bone formation, bone mass, and bone strength in a rat model of postmenopausal osteoporosis.Journal of Bone & Mineral Research the Official Journal of the American Society for Bone &Mineral Research, 2009, 24(4):578-588.
[14] Durlej M, Knapczyk-Stwora K, Duda M, et al. The expression of FSH receptor (FSHR) in the neonatal porcine ovary and its regulation by flutamide. Reproduction in Domestic Animals,2011, 46(3):377-384.
[15] Haberman S, Race G J. Application of immunofluorescence to study of human tumors by rabbit antibody production to nucleoprotein of human malignant tissues. Texas Medicine, 1968, 64(5):45.
[16] Heukers R, Henegouwen P M, Oliveira S. Nanobody-photosensitizer conjugates for targeted photodynamic therapy. Nanomedicine:Nanotechnology, Biology and Medicine, 2014,10(7):1441-1451.
[17] Kalu D N. The ovariectomized rat model of postmenopausal bone loss. Bone & Mineral, 1992,15(3):175-191.
[18] Liu S P, Liao E Y, Hanwen W U, et al. Comprehensive assessment of the ovariectomized rat model of postmenopausal osteoporosis. Bulletin of Hunan Medical University, 2001, 26(2):111-114.
[19] Pujol J P, Chadjichristos C, Legendre F, et al. Interleukin-1 and transforming growth factor-β1 as crucial factors in osteoarthritic cartilage metabolism. Connective Tissue Research, 2008,49(3-4):293-297.

[1]	丁艳, 杨隽, 司书毅, 阮力. Statin类药物促成骨细胞BMP-2表达机理[J]. 中国生物工程杂志, 2005, 25(4): 18-21.
[2]	钟辉, 彭英芳, 李平, 袁志刚, 杨晓莉, 马清钧. 鲑鱼降钙素与骨生长肽融合基因的克隆及其在毕赤酵母中的表达[J]. 中国生物工程杂志, 2002, 22(5): 79-82.
[3]	钟辉, 彭英芳, 李平, 袁志刚, 杨晓丽, 马清钧. 毕赤酵母中表达的鲑鱼降钙素与骨生长肽融合蛋白的活性检测[J]. 中国生物工程杂志, 2002, 22(4): 84-88.
[4]	王欣荣, 应汉杰, 欧阳平凯. 骨质疏松症的发病机理及其治疗[J]. 中国生物工程杂志, 2001, 21(3): 54-56.

Viewed

Full text

Abstract

Cited

Shared

Discussed