本文選題：尿道下裂　切入點(diǎn)：成纖維細(xì)胞　出處：《福建醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 1.體外觀察尿道下裂患兒陰莖包皮內(nèi)板組織成纖維細(xì)胞生長(zhǎng)特點(diǎn)，探討成纖維細(xì)胞生長(zhǎng)發(fā)育異常與尿道下裂發(fā)生的關(guān)系。 2.通過(guò)EGF和T干預(yù)尿道下裂包皮成纖維細(xì)胞，體外試驗(yàn)觀察EGF和T對(duì)其AR活化水平的影響，探究尿道下裂的發(fā)病機(jī)制。 方法： 1.組織塊法原代培養(yǎng)尿道下裂患兒的陰莖包皮內(nèi)板組織成纖維細(xì)胞及細(xì)胞增殖試驗(yàn)檢測(cè)細(xì)胞增值情況，流式細(xì)胞術(shù)分析細(xì)胞周期。 2.免疫細(xì)胞化學(xué)技術(shù)檢測(cè)EGF和T干預(yù)后尿道下裂成纖維細(xì)胞AR活化水平的變化。 結(jié)果： 1.細(xì)胞原代培養(yǎng)試驗(yàn)發(fā)現(xiàn)近端型尿道下裂組成纖維細(xì)胞移出組織塊的時(shí)間明顯長(zhǎng)于中遠(yuǎn)端型組和對(duì)照組（P值均0.05），同時(shí)細(xì)胞生長(zhǎng)融合成束所需時(shí)間也緩于中遠(yuǎn)端型組和對(duì)照組（P值均0.001）； 2.細(xì)胞增殖試驗(yàn)和細(xì)胞周期分析，顯示近端型組尿道下裂受檢成纖維細(xì)胞增殖水平明顯低于中遠(yuǎn)端型組和對(duì)照組（P值均0.001）；中遠(yuǎn)端型低于對(duì)照組（P0.05）。 3.在生理濃度睪酮（3×10~(-8)mol/L）的作用下，近端型尿道下裂組的成纖維細(xì)胞與其他兩組比較AR的活化明顯下降（P0.001）。 4.在睪酮濃度為3×10~(-6)mol/L時(shí)，三個(gè)組AR活化存在明顯差異（P0.001），正常組均數(shù)中遠(yuǎn)端型組均數(shù)近端型組均數(shù)。 5.在EGF和生理濃度睪酮干預(yù)下，近端型尿道下裂組的成纖維細(xì)胞AR的活化程度升高最為明顯與正常組以及中遠(yuǎn)端型組有明顯差異（P0.001），中遠(yuǎn)端型組AR的活化升高的水平與對(duì)照組也具有差異（P=0.020.05）。 結(jié)論： 1.尿道下裂患兒生殖器成纖維細(xì)胞增殖能力下降，可能是尿道下裂，，尤其是近端型尿道下裂發(fā)病的重要因素之一 2.尿道下裂成纖維細(xì)胞AR的表達(dá)與活化均存在異常，比正常對(duì)照組活化水平明顯下降，尿道下裂的發(fā)生可能與EGF作用缺陷導(dǎo)致T的生物學(xué)活性不足以及AR表達(dá)異常有關(guān)。 3.在睪酮濃度為3×10~(-6)mol/L時(shí)，成纖維細(xì)胞的增殖能力達(dá)到最佳，AR活化達(dá)到該條件下最大水平，而聯(lián)合EGF與睪酮可以進(jìn)一步增強(qiáng)成纖維細(xì)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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R726.92

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 周娟;王浩;彭御冰;陳其;達(dá)駿;王忠;馬利民;;ATF3和CTGF基因在尿道下裂患者包皮中的表達(dá)及雌激素調(diào)控研究[J];中華男科學(xué)雜志;2009年12期

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