神经干细胞移植后在视神经损伤大鼠视网膜内表达BDNF的研究

Expression of BDNF after neural stem cell transplantation in the retina of the Sprague-Dawley rats with optic nerve injury

目的探讨神经干细胞(NSC)移植入视神经损伤(ONI)的SD大鼠玻璃体下腔后在视网膜内表达脑源性神经营养因子(BDNF)的情况,为进一步探索NSC移植治疗视神经损伤提供实验依据。方法体外培养NSC,其上清液采用酶联免疫吸附实验(ELISA)定量分析BDNF含量。取34只SD大鼠,其中4只作为正常对照,另外30只制作成右眼ONI模型并随机等分为N组和P组。ONI术后随即向N组大鼠右眼玻璃体下腔注入定量NSC,P组注入等量PBS,并采用半定量RT-PCR方法检测正常大鼠视网膜及N组、P组大鼠术后第3天、1周、2周、3周、4周时视网膜BDNFmRNA的表达水平,行统计学分析。结果正常视网膜内可见BDNF表达;N组与P组大鼠视网膜内表达BDNF的量除第1周差异无统计学意义外,余时间段N组均高于P组。结论NSC能促使视神经损伤大鼠视网膜高表达BDNF,NSC移植治疗视神经损伤值得进一步深入研究。

Objective To explore the expression of brain-derived neurotrophic factor （BDNF） in the retina after transplantation of neural stem calls （NSCs） into the inferior vitreous cavity of the Sprague-Dawley （SD） rats with optic nerve injury （ONI）, and provide experimental evidence for NSCs transplantation therapy for ONI. Methods At first, NSCs was cultured in vitro, and the supematant liquid was detected of the BDNF by ELISA quantitative analysis. Secondly, thirty-four SD rats were divided into normal control group with 4 rats, and groups N and P with 15 rats each. Immediately after ONI, cultured NSCs were injected into the right vitreous body in Group N rats and the same dose of PBS was injected into the right vitreous in group P rats. Semi-quantitative RT-PCR was used to detect mRNA expression of BDNF in the normal retinas and retinas 3 days, 1 week, 2, 3 and 4 weeks after the operation in groups N and P, with 3 rats each time in every group. The data were analyzed statistically. Results The expression of BDNF mRNA was observed in normal retinas. The expression level of BDNF mRNA in group N was almost all higher than that in group P in every time period, except the first week with no statistic difference. Conclusion NSCs transplanted in retinas of ONI rats can secrete large quantities of BDNF. It is worth deep exploring NSCs transplantation therapy for ONI.