中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (33): 6129-6132.doi: 10.3969/j.issn.1673-8225.2010.33.012

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

重组pEGFP-N1-IGF-1质粒体内转染骨质疏松大鼠的可行性

陈  剑，陈允震，刘海春，丁  岩， 赵  华   

  1. 山东大学齐鲁医院骨创伤科，山东省济南市  250012
  • 出版日期:2010-08-13 发布日期:2010-08-13
  • 通讯作者: 陈允震，硕士，教授，山东大学齐鲁医院骨创伤科，山东省济南市 250012 cyz112@yahoo.com.cn
  • 作者简介:陈 剑★，男，1983年生，山东省平邑县人，汉族，山东大学在读硕士，医师，主要从事脊柱外科、骨质疏松方面的研究。 chnjan@qq.com
  • 基金资助:

    山东省科技厅攻关课题(2008GG10002057)，课题名称：骨胶原在骨质疏松发生及其治疗中作用机制的分子生物学研究。

Feasibility of recombinant plasmid pEGFP-N1-IGF-1 transfection in an osteoporosis rat

Chen Jian, Chen Yun-zhen, Liu Hai-chun, Ding Yan, Zhao Hua   

  1. Department of Orthopaedics, Qilu Hospital, Shandong University, Jinan   250012, Shandong Province, China
  • Online:2010-08-13 Published:2010-08-13
  • Contact: Chen Yun-zhen, Master, Professor, Department of Orthopaedics, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China cyz112@yahoo.com.cn
  • About author:Chen Jian★, Studying for master’s degree, Physician, Department of Orthopaedics, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China chnjan@qq.com
  • Supported by:

    the Tackle Key Program of Department of Science and Technology of Shandong Province, No. 2008GG10002057*

PDF

363

被引次数

3

摘要：

背景：胰岛素样生长因子1水平的下降与骨质疏松密切相关，目前应用基因技术是骨质疏松等骨代谢疾病治疗的发展方向，但病毒载体可能引起严重免疫反应，质粒重组体pEGFP-N1-IGF-1活体转染骨质疏松大鼠可能成为一种有效的治疗手段。
目的：观察质粒重组体pEGFP-N1-IGF-1转染骨质疏松大鼠中胰岛素样生长因子1的表达。
方法：雌性SD大鼠随机分为假手术组、pEGFP-N1组和pEGFP-N1-IGF-1组。后2组予双侧卵巢切除。术后12周，3组依次给予生理盐水、pEGFP-N1载体和质粒重组体pEGFP-N1-IGF-1分别复合脂质体转染。转染后48 h荧光活体成像及肝脏切片观察，定期测定血清胰岛素样生长因子1质量浓度。
结果与结论：转染后pEGFP-N1组和pEGFP-N1-IGF-1组大鼠均可见全身大部出现明显荧光表达，以肝脏及尾部为著，肝脏切片镜下观察可见明显荧光表达。去卵巢大鼠血清胰岛素样生长因子1质量浓度显著降低(P < 0.05)；pEGFP-N1-IGF-1转染使去卵巢大鼠血清胰岛素样生长因子1质量浓度明显升高(P < 0.05)，而后随时间延长而逐渐降低(P < 0.05)。结果证明质粒重组体pEGFP-N1-IGF-1能够在骨质疏松大鼠体内成功转染并表达胰岛素样生长因子1蛋白，为应用胰岛素样生长因子1基因治疗骨质疏松症提供了理论基础。

关键词: 胰岛素样生长因子1, 质粒, 脂质体, 活体成像, 骨质疏松, 大鼠, 骨组织工程

Abstract:

BACKGROUND: The decline of insulin-like growth factor-1 (IGF-1) levels has been proven to be an important reason leading to osteoporosis. Gene therapy for osteoporosis and other bone metabolic diseases has becoming a direction of the research, but the viral vector may cause serious immune response. Therefore, the recombinant plasmid pEGFP-N1-IGF-1 in vivo transfection of osteoporosis rats may be a more effective treatment way.
OBJECTIVE: To study the IGF-1 expression during recombinant plasmid pEGFP-N1-IGF-1 in the osteoporosis rats in vivo.
METHODS: Female Sprague Dawley rats were randomly assigned to sham-surgery group, pEGFP-N1 group and pEGFP-N1-IGF-1 group. Bilateral ovariotomy was performed in pEGFP-N1 group and pEGFP-N1-IGF-1 group. At 12 weeks following surgery, the three groups were given saline, pEGFP-N1 vector and recombinant plasmid pEGFP-N1-IGF-1 complex liposomes hydrodynamic injection, respectively. Fluorescence in vivo imaging and liver biopsy were observed at 48 hours following transfection. Serum concentrations of IGF-1 were determined at regular intervals.
RESULTS AND CONCLUSION: After transfection, rats of pEGFP-N1 group and pEGFP-N1-IGF-1 group developed noticeable fluorescence expression not only in the whole body but also liver biopsy, especially in the liver and tail. IGF-1 levels were significantly decreased in rats undergoing ovariotomy (P < 0.05). The expression level of serum IGF-1 in rats of pEGFP-N1-IGF-1 group were significantly increased (P < 0.05), and then reduced over time (P < 0.05). Results indicated that recombinant plasmid pEGFP-N1-IGF-1 can be successfully transfected and expressed IGF-1 protein in osteoporosis rats, which can be a substantial foundation for further application of IGF-1 gene therapy to osteoporosis.

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