摘要
目的探讨二氢睾酮(dihydrotestosterone,DHT)对损伤后大鼠视网膜神经节细胞-5(retinal ganglion cell-5,RGC-5)神经突起再生及RhoA和神经突蛋白(neuritin)表达的影响。方法 RGC-5经十字孢碱诱导分化后,将细胞分为正常对照组、模型组[用连二亚硫酸钠(Na2S2O4)制备大鼠RGC-5神经突起氧糖剥夺/复氧损伤模型]和DHT干预组(终浓度分别为1、10、100 nmol/L),采用MTT比色法测定各组细胞的存活率;显微镜下观察神经突起长度和数量;RT-PCR和Western blot法分别检测RGC-5中RhoA和Neuritin基因mRNA的转录水平及蛋白的表达水平。结果与模型组比较,10和100 nmol/L DHT干预组及正常对照组的细胞存活率和神经突起数量均明显上升(P均<0.05);1、10和100 nmol/L DHT干预组及正常对照组的神经突起长度、Neuritin基因mRNA转录水平和蛋白表达量均明显增加(P均<0.05),RhoA基因mRNA转录水平及蛋白表达量均明显减少(P均<0.05)。结论 RhoA和Neuritin可能是影响RGC-5神经突起再生的重要因子,DHT可能通过调节RhoA和Neuritin的表达,从而发挥神经保护作用。
Objective To investigate the effect of dihydrotestosterone（DHT）on the neurite regeneration and expressions of RhoA and Neuritin in rat retinal ganglion cells-5（RGC-5）. Methods Differentiation of RGC-5 was induced by stauros- porine and used for establishment of oxygen-glucose deprivation / reoxygenation neurite injury model serving sodium hy- drosulfite as an anoxic agent. Differentiated cells were divided into normal control, model control and three test groups. The cells in test groups 1, 2 and 3 were treated with 1, 10 and 100 nmol / L DHT respectively, while those in normal control and model control groups were untreated. The cells were determined for survival rate by MTT assay, observed for length and number of neurite by microscopy, analyzed for transcription levels of RhoA and Neuritin mRNAs by RT-PCR, and for expression levels of RhoA and Neuritin proteins by Western blot. Results As compared with those in model control group, the survival rates and numbers of neuritie of cells in test groups 2 and 3 as well as normal control group increased significantly（each P〈 0. 05）;however, the lengths of neurite as well as mRNA transcription and protein expression lev- els of Neuritin in three test groups and normal control group increased significantly（each P〈 0. 05）, while the mRNA transcription and protein expression levels of RhoA decreased significantly（each P〈 0. 05）. Conclusion Rho A and Neuritin might be important influencing factors of neurite regeneration in RGC-5. DHT might give play to its neuropro- tection by regulating the expressions of RhoA and Neuritin.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第2期207-211,共5页
Chinese Journal of Biologicals
