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重组质粒 pEGFP-N1-IGF-1 转染在减轻去卵巢大鼠骨质疏松症中的作用。

Role of recombinant plasmid pEGFP-N1-IGF-1 transfection in alleviating osteoporosis in ovariectomized rats.

机构信息

Department of Orthopaedics, Qilu Hospital of Shandong University, Jinan, 250012, China.

出版信息

J Mol Histol. 2013 Oct;44(5):535-44. doi: 10.1007/s10735-013-9498-3. Epub 2013 Mar 19.

Abstract

Decreased levels of serum insulin-like growth factor-1 (IGF-1) have been proven to cause osteoporosis. Gene transfer of IGF-1 offers an attractive technology to treat skeletal metabolic disorders including osteoporosis, but the viral vectors are limited by their high antigenicity and immune response. Our purpose was to investigate the expression of a non-invasive vector, recombinant plasmid enhanced green fluorescent protein-N1 (pEGFP-N1) that transferred IGF-1 gene into ovariectomized (OVX) rats in vivo and evaluate the effect of this therapy on osteoporosis. OVX or sham operations were performed in 60 female, 7-month-old unmated SD rats. 12 weeks after OVX operation, the vectors were transfected to the 10-month-old rats and experimental data were detected from 48 h to 7 week after transfection. Our results showed that remarkable expression of fluorescence and serum IGF-1 was observed in the rats transfected by recombinant plasmids, indicating that IGF-1 gene was successfully transferred to OVX rats by injecting the vector through hydrodynamic method via the tail vein. The bone metabolism index including serum alkaline phosphatase, the histomorphometric parameters of lumbar vertebra including trabecular area percentage, trabecular thickness, trabecular number and trabecular separation, and the bone mineral density (BMD) and biomechanical parameters of lumbar vertebra including BMD, maximum condensing force, crushing strength in OVX rats transfected by pEGFP-N1-IGF-1 were improved remarkably compared with OVX+pEGFP-N1 rats, indicating that the transfection of recombinant plasmid pEGFP-N1-IGF-1 played a significant role in alleviating osteoporosis in rats induced by OVX. This encouraged a potential approach of IGF-1 gene therapy to the treatment of osteoporosis.

摘要

血清胰岛素样生长因子-1(IGF-1)水平降低已被证明可导致骨质疏松症。IGF-1 的基因转移为治疗包括骨质疏松症在内的骨骼代谢紊乱提供了一种有吸引力的技术,但病毒载体受到其高抗原性和免疫反应的限制。我们的目的是研究一种非侵入性载体,即重组质粒增强型绿色荧光蛋白-N1(pEGFP-N1),该载体将 IGF-1 基因转移到体内去卵巢(OVX)大鼠中,并评估该治疗方法对骨质疏松症的影响。在 60 只 7 月龄未交配的 SD 雌性大鼠中进行 OVX 或假手术。OVX 手术后 12 周,将载体转染至 10 月龄大鼠,在转染后 48 小时至 7 周检测实验数据。我们的结果表明,重组质粒转染的大鼠中观察到荧光和血清 IGF-1 的显著表达,表明通过尾静脉注射载体的水力方法成功地将 IGF-1 基因转染到 OVX 大鼠中。骨代谢指标包括血清碱性磷酸酶,腰椎的组织形态计量学参数包括小梁面积百分比、小梁厚度、小梁数量和小梁分离度,以及 OVX+pEGFP-N1-IGF-1 转染大鼠的腰椎骨密度(BMD)和生物力学参数,包括 BMD、最大压缩力和压缩强度,均显著改善,表明重组质粒 pEGFP-N1-IGF-1 的转染对 OVX 诱导的大鼠骨质疏松症有明显的缓解作用。这为 IGF-1 基因治疗治疗骨质疏松症提供了一种潜在的方法。

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