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| 外源性NGF对转基因CGRP干细胞移植治疗骨质疏松大鼠的影响 |
| The effect of exogenous NGF on the treatment of osteoporosis with CGRP gene transgenic MSCs transplantation in rats |
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| DOI：10.3969/j.issn.1006.7108.2015.12.005 |
| 中文关键词: 降钙素基因相关肽；骨髓干细胞移植；神经生长因子；骨质疏松 骨代谢 |
| 英文关键词:Calcitonin gene related peptide Marrow stromal cells Nerve growth factor Osteoporosis Bone metabolism |
| 基金项目:广东省自然科学基金（9151051501000054);广东省科技计划项目 (2011B031800064) |
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| 中文摘要: |
| 目的 研究外源性神经生长因子（nerve growth factor,NGF)对转基因降钙素基因相关肽（calcitonin gene related peptide, CGRP)骨髓干细胞（mesenchymal stem cells，MSCs)移植治疗骨质疏松大鼠的影响。方法 构建CGRP基因表达载体,转染制备转基因CGRP-MSCs靶细胞;60只雌性SD大鼠骨质疏松模型随机均分为空白对照组（A组）、单纯MSCs移植治疗组（B 组）、CGRP-MSCs移植治疗组（C组）,CGRP-MSCs + NGF移植组（D组）。在治疗lw、2w、3w后，实时定量荧光PCR和Western blot检测3个时间段每组大鼠股骨骨组织中CGRP基因和蛋白的相对表达水平，评价各组疗效。结果 与A组比较,3个时间点，C组和D组中CGRP表达水平均显著上调（P<0.01);治疗lw后，C组和D组上调水平差异明显（P<0.01)，D组低于C 组；治疗2w后，C组和D组之间仍保持显著差异（P <0.05)，D组仍低于C组；治疗3w后，C组和D组上调水平差异明显缩 小,二者差异无统计学意义（P>0.05)。CGRP基因和蛋白相对表达水平在各组检测结果差异一致，同上。结论 在转基因 CGRP-MSCs移植治疗骨质疏松大鼠中联合应用外源性NGF,无累计效应，并可能抑制大鼠股骨骨组织中CGRP的表达，表明外源NGF于转基因CGRP-MSCs移植治疗骨质疏松症中可能并无促进作用，至少在CGRP表达层面无促进作用，而单纯的转基因CGRP-MSCs移植治疗骨质疏松大鼠疗效优于联合应用外源性NGF治疗方案。 |
| 英文摘要: |
| Objective To investigate the effect of exogenous nerve growth factor (NGF) on osteoporotic rats under therapy with mesenchymal stem cell (MSC) transfected with calcitonin gene related peptide (CGRP) gene. Methods Vectors express CGRP were constructed and transfected to CGRP-MSCs target cells. Sixty female osteoporotic SD rats were randomly divided into the control group (group A), simple MSCs transplantation treatment group ( group B )，CGRP transgenic MSCs ( CGRP-MSCs) transplantation treatment group (group C)，and CGRP-MSCs transplantation with exogenous NGF (CGRP-MSCs + NGF) treatment group (group D). At 1, 2, and 3 weeks after the treatment，CGRP expression levels in the femurs was measured using real-time PCR and Western blotting. The efficacy was compared between each group. Results Compared with control group, CGRP expression in Group C and D were both significantly up-regulated (P <0. 01) at all time. After 1 - week treatment, the difference between group C and D was significant (P < 0. 01). The expression in group D was significantly lower than that in the group C. After2 - week treatment, the difference between group C and D was still significant (P<0. 05)，and the expression in group D was still significantly lower than that in group C. After 3-week treatment, the up-regulation difference in group D and C became narrower, and the difference was not statistically significant (P > 0. 05). The results of CGRP gene and protein expression were consistent in all groups. Conclusion This study suggests that additional application of exogenous NGF may inhibit CGRP expression when CGRP gene transgenic MSCs transplantation therapy is applied in osteoporosis rat model，indicating that additional application of exogenous NGF has no benefit to the effect of CGRP, at least on CGRP gene expression level. CGRP gene transgenic MSCs transplantation therapy without exogenous NGF is better than additional application of exogenous NGF. |
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