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| 过表达COLIA1 cDNA腺病毒感染TT型成骨细胞株后 对其生物学性能影响的研究 |
| The effect of over-expression of COLIAlcDNA in adenovirus-infected TT osteoblasts on their biochemical functions |
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| DOI： |
| 中文关键词: I型胶原 基因多态性 4997位点 成骨细胞 |
| 英文关键词:Type I collagen Polymorphism -L997 site Osteoblast |
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| 中文摘要: |
| 目的本实验前期研究发现I型胶原a1链基因4997G—T纯合突变可降低成骨细胞生物学性能。现利用基因重组技 术提高TT型成骨细胞COLIA1基因mRNA的表达水平，观察能否逆转COLIA1基因4997G—T突变对TT型成骨细胞生物学 性能的影响。方法构建过表达COLIA1基因的腺病毒载体，并感染TT型成骨细胞。比较感染后TT型成骨细胞I型胶原a1 链mRNA的表达水平、I型胶原的含量、细胞基质钙含量以及钙结节数量的差异。结果过表达COLIA1基因的腺病毒载体感 染后，TT型成骨细胞I型胶原a1链mRNA的表达水平、I型胶原的含量、细胞基质钙含量以及钙结节数量较未感染组及空病 毒组有显著提高，差异有统计学意义。结论利用腺病毒载体提高TT型成骨细胞的I型胶原a1链mRNA的表达水平，可增 加I型胶原的含量、细胞基质钙含量以及钙结节数量。TT型成骨细胞I型胶原合成的减少导致细胞外基质减少，导致钙质沉 着部位不足可能是4997G/T患者BMD降低的原因。 |
| 英文摘要: |
| Objective Based on the findings that the homozygotic mutation of G to T in a1 chain-L997 could lower the biochemical functions in osteoblasts in our preliminary study，to enhance COLIA1 mRNA expression level using recombinant DNA technology in TT genotype osteoblasts，and to observe whether it could reverse the effect of COLIA1-L997 G^T mutation on the biology performance in TT genotype osteoblasts. Methods The COLIA1 cDNA over-expression adenovirus vector was constructed and used to infect TT genotype osteoblasts. The expression level of COLIA1 mRNA，the content of type I collagen，the content of the calcium in cell matrix， and the amount of calcium nodules in infected TT genotype osteoblasts were detected and analyzed. Results After the infection of COLIA1 cDNA over-expression adenovirus vector in TT genotype osteoblasts，the expression level of COLIA1 mRNA，the content of type I collagen，the content of the calcium in cell matrix，and the amount of calcium nodules increased significantly compared with those in uninfected group and non-virus group (P < 0. 05). Conclusion The expression level of COLIA1 mRNA increases after the infection of COLIA1 cDNA over-expression adenovirus vector in TT genotype osteoblasts， leading to an improvement of the content of type I collagen， calcium in cell matrix， and the amount of calcium nodules. The lower content of synthesized type I collagen in TT genotype osteoblasts results in the decrease of extracellular matrix， leading to insufficient site for calcium deposition，which maybe the cause of low BMD in TT genotype patients. |
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