人睫状神经营养因子结构和功能的研究

A structure function analysis of the human ciliary neurotrophic factor

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 recombinant human ciliary neurotrophic factor (hCNTF) of high biological activity and study its biological function. Methods:The human CNTF gene was expressed in E.coli.. Sciatic nerve sectioned in the rat was sutured into silicone chamber, CNTF was injected into the silicone chambers and subcutis, and HRP retrograde tracing was used to observe the soma of the regenerated axons pass. Results: The results showed that the α helixes in CNTF were critical for the biological activity and the flexible C terminus of human CNTF was not essential. Our data also indicated that the middle and tail part of the D helix might play crucial roles in the biological functions of CNTF. Compared with control group, the number of labeled spinal soma was only significantly increased in CNTF group. Conclusion: The study provides theory basis for preparation of CNTF having high activity and low side effect. The CNTF can enhance motor axon.