胶质细胞源性神经营养因子对体外培养大鼠精原干细胞PLZF和c-Kit表达的影响  

Effects of glial cell line-derived neurotrophic factor on the proliferation and differentiation of PLZF and e-Kit of spermatogonial stem cells of rats in vitro

作　　者：夏伟[1] 曾甫清[2] 叶哲伟[3] 白杨[2] 

机构地区：[1]华中科技大学同济医学院计划生育研究所,武汉430030 [2]华中科技大学同济医学院协和医院泌尿外科,武汉430030 [3]华中科技大学同济医学院协和医院骨科,武汉430030

出　　处：《中华泌尿外科杂志》2010年第4期223-226,共4页Chinese Journal of Urology

基　　金：国家自然科学基金资助项目（30371424）

摘　　要：目的探讨胶质细胞源性神经营养因子（GDNF）对大鼠原代培养的精原干细胞（SSC）早幼粒细胞白血病锌指蛋白（PLZF）、原癌基因c-Kit基因转录及表达的影响。方法采用含不同浓度GDNF（0、10、50、100ng／m1）的DMEM培养液原代培养大鼠SSC。RT-PCR检测SSC中PLZFmRNA、c-Kit mRNA水平，蛋白质印迹法检测PLZF、c-Kit蛋白表达情况。结果对照组和GDNF10ng／ml组细胞随着培养时间延长，生长速度无明显变化，而GDNF50和100ng／ml组细胞生长速度明显加快。对照组PLZF和c-KitmRNA表达量分别为0．28±0．13、0．65±0．21，GDNF10ng／ml组分别为0．27±0．14、0．62±0．19，与对照组比较差异无统计学意义（P〉0．05）；GDNF50ng／ml组分别为0．64±0．28、0．34±0．15，与对照组比较差异均有统计学意义（P〈0．05）；GDNF100ng／ml组分别为0．68±0．27、0．28±0．18，与50ng／ml组比较差异无统计学意义（P〉0．05）。对照组PLZF和c-Kit蛋白表达量分别为0．34±0．13、0．72±0．27；GDNF10ng／ml组分别为0．38士0．18、0．69土0．26，与对照组比较差异无统计学意义（P〉0．05）；GDNF50ng／ml组分别为0．68±0．26、0．35±0．15，与对照组比较差异有统计学意义（P〈0．05）;GDNF100ng／ml组分别为0．70±0．27、0．32±0．11，与50ng／ml组比较差异无统计学意义（P2〉0．05）。结论随着GDNF浓度增加，SSC增殖水平升高，而分化受抑制。GDNF对SSC增殖和分化有一定的调控作用。Objective To investigate the effects of glial cell line-derived neurotrophic factor（GDNF） on the gene transcriptions and expressions of promyelocytic leukaemia zinc finger （PLZF） and c-Kit in spermatogonial stem ceils （SSC）. Methods SSC was cultured in DMEM plus different concentrations of GDNF（0, 10, 50, 100 ng/ml）. Quantitative reverse transcription PCR was used to detect the mRNA of PLZF and c-Kit. The expressions of PLZF and c-Kit protein were detected by Western blot assay. Results With the control group and 10 ng/ml of GDNF group cell growth has no obvious influence, but with 50 ng/ml of GDNF group and 100 ng/ml of GDNF group cell growth was enhanced significantly. In the control group, the PLZF mRNA was 0. 28±0. 13 and c-Kit mRNA was 0. 65±0. 21. In the 10 ng/rnl of GDNF group, PLZF mRNA was 0.27±0.14 and c-Kit mRNA was 0.62±0. 19. Compared with the control group, 10 ng/ml of GDNF group had not influence on PLZF and c-Kit mRNA. In the 50 ng/ml of GDNF group PLZF mRNA was 0.64±0. 28 and c-Kit mRNA was 0. 34±0.15. Compared with the control group, 50 ng/ml of GDNF enhanced the transcription of PLZF mRNA （P〈0. 05）, decreased the transcription of c-Kit mRNA （P〈0. 05）. In the 100 ng/ml of GDNF group, PLZF mRNA was 0. 68±0. 27 and c-Kit mRNA was 0. 28±0. 18. In the 100 ng/ml of GDNF group, there was no difference compared with 50 ng/ml of GDNF. In the control group, PLZF protein was 0. 34±0. 13 and c-Kit protein was 0.72±0. 27. In 10 ng/ml of GD NF group, PLZF protein was 0. 38±0.18 and c-Kit protein was 0. 69±0. 26. Compared with the control group, 10 ng/ml of GDNF had no influence on PLZF and c-Kit protein. In 50 ng/ml of GDNF group, PLZF protein was 0.68±0. 26 and c-Kit protein was 0. 35±0. 15. 50 ng/ml of GDNF enhanced the expression of PLZF protein （P〈0. 05）, decreased the expression of c-Kit protein（P〈0.05）. In 100 ng/ml of GDNF group PLZF protein was 0. 70-1-0.27 and c-Kit protein was 0. 32±0.11, 100 ng/ml of GDNF had no influence on PLZF and c-Kit pro

关 键 词：精原细胞 胶质细胞源性神经营养因子 早幼粒细胞白血病锌指蛋白 原癌基因蛋白质 C-KIT 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]