尿道下裂患儿阴茎包皮内板组织成纤维细胞生长特点及表皮生长因子对其AR活化水平的影响

发布时间：2018-03-08 22:13

本文选题：尿道下裂　切入点：成纤维细胞　出处：《福建医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的： 1.体外观察尿道下裂患儿阴茎包皮内板组织成纤维细胞生长特点，探讨成纤维细胞生长发育异常与尿道下裂发生的关系。 2.通过EGF和T干预尿道下裂包皮成纤维细胞，体外试验观察EGF和T对其AR活化水平的影响，探究尿道下裂的发病机制。方法： 1.组织块法原代培养尿道下裂患儿的阴茎包皮内板组织成纤维细胞及细胞增殖试验检测细胞增值情况，流式细胞术分析细胞周期。 2.免疫细胞化学技术检测EGF和T干预后尿道下裂成纤维细胞AR活化水平的变化。结果： 1.细胞原代培养试验发现近端型尿道下裂组成纤维细胞移出组织块的时间明显长于中远端型组和对照组（P值均0.05），同时细胞生长融合成束所需时间也缓于中远端型组和对照组（P值均0.001）； 2.细胞增殖试验和细胞周期分析，显示近端型组尿道下裂受检成纤维细胞增殖水平明显低于中远端型组和对照组（P值均0.001）；中远端型低于对照组（P0.05）。 3.在生理浓度睾酮（3×10~(-8)mol/L）的作用下，近端型尿道下裂组的成纤维细胞与其他两组比较AR的活化明显下降（P0.001）。 4.在睾酮浓度为3×10~(-6)mol/L时，三个组AR活化存在明显差异（P0.001），正常组均数中远端型组均数近端型组均数。 5.在EGF和生理浓度睾酮干预下，近端型尿道下裂组的成纤维细胞AR的活化程度升高最为明显与正常组以及中远端型组有明显差异（P0.001），中远端型组AR的活化升高的水平与对照组也具有差异（P=0.020.05）。结论： 1.尿道下裂患儿生殖器成纤维细胞增殖能力下降，可能是尿道下裂，，尤其是近端型尿道下裂发病的重要因素之一 2.尿道下裂成纤维细胞AR的表达与活化均存在异常，比正常对照组活化水平明显下降，尿道下裂的发生可能与EGF作用缺陷导致T的生物学活性不足以及AR表达异常有关。 3.在睾酮浓度为3×10~(-6)mol/L时，成纤维细胞的增殖能力达到最佳，AR活化达到该条件下最大水平，而联合EGF与睾酮可以进一步增强成纤维细胞AR活化的水平。
[Abstract]:Objective:. 1. To observe the growth characteristics of fibroblasts in the prepuce of penis in children with hypospadias in vitro, and to explore the relationship between the abnormal growth and development of fibroblasts and the occurrence of hypospadias. 2. The effects of EGF and T on AR activation of hypospadias prepuce fibroblasts were observed by EGF and T in vitro to explore the pathogenesis of hypospadias. Methods:. 1. The primary culture of hypospadias in children with hypospadias by tissue mass method was used to detect the proliferation of fibroblasts and cell proliferation in the prepuce of penis. Flow cytometry was used to analyze the cell cycle. 2. The changes of AR activation in hypospadias fibroblasts after EGF and T intervention were detected by immunocytochemistry. Results:. 1. In the primary culture test, it was found that the time of removal of fibroblasts from the proximal hypospadias was significantly longer than that of the middle and distal type and control group (P = 0.05), and the time of cell growth and fusion into bundle was also slower than that of the middle and distal type. P value of both group and control group were 0.001g; 2. Cell proliferation test and cell cycle analysis showed that the proliferative level of hypospadias fibroblasts in proximal type group was significantly lower than that in middle and distal type group and control group (P value 0.001), and that in the middle distal type group was lower than that in control group (P 0.05). 3. The activation of AR in the proximal hypospadias group was significantly lower than that in the other two groups under the physiological concentration of testosterone 3 脳 10 ~ (-8) mol 路L ~ (-1). 4. When the testosterone concentration was 3 脳 10 ~ (-6) mol / L, there was a significant difference in AR activation among the three groups (P 0.001). 5. Under the intervention of EGF and physiological concentration testosterone, The activation of AR in fibroblasts in the proximal hypospadias group was significantly higher than that in the normal group and the middle and distal type group (P 0.001), and the level of AR activation in the middle and distal type group was also significantly higher than that in the control group (P 0.020.05). Conclusion:. 1. Hypospadias in children with hypospadias may be one of the important factors in the pathogenesis of hypospadias, especially proximal hypospadias. 2. The expression and activation of AR in hypospadias fibroblasts were abnormal, which was significantly lower than that in normal controls. The occurrence of hypospadias might be related to the deficiency of T activity and the abnormal expression of AR in hypospadias due to the deficiency of EGF. 3. When the concentration of testosterone was 3 脳 10 ~ (-1) -6 渭 mol / L, the proliferation ability of fibroblasts reached the best level, and the combination of EGF and testosterone could further enhance the level of AR activation of fibroblasts.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.92

【参考文献】