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目的:制备高活性的重组人睫状神经营养因子(hCNTF),并研究其生物学功能。方法:应用大肠杆菌表达hCNTF,用片段插入和缺失法研究其结构与功能关系;切断大鼠坐骨神经,局部及皮下给予CNTF,应用辣根过氧化物酶逆行追踪技术显示再生轴突通过修复部位的胞体。结果:hCNTF分子中α-螺旋结构的维持对其生物活性十分重要,C端松散结构对其生物活性贡献不大,D螺旋中后段可能与生物活性有密切关系;与对照组相比,CNTF组脊髓标记胞体显著增加。结论:本研究为获得高活性、低副作用的CNTF提供了理论依据;所获hCNTF能明显促进周围神经运动轴突的再生。
Objective: To prepare highly active recombinant human ciliary neurotrophic factor (hCNTF) and study its biological function. Methods: The hCNTF was expressed in Escherichia coli. The structure and function of the regenerated axon were studied by fragment insertion and deletion. The sciatic nerve was excised and the CNTF was administered locally and subcutaneously. HRP reverse-tracing technique was used to show that the regenerated axon passes through the repair site Cell body. Results: The maintenance of α-helical structure in hCNTF molecule was very important to its biological activity. The loose structure of C-terminal did not contribute much to the biological activity of hCNTF molecule. The middle and late D helix might be closely related to the biological activity. Compared with the control group, CNTF Group of spinal cord labeled cell body significantly increased. Conclusion: This study provides a theoretical basis for obtaining high activity, low side effects of CNTF; obtained hCNTF can significantly promote the regeneration of motor axons.