【摘要】 目的明确PPP2R2C蛋白在不同发育阶段的小鼠睾丸中的表达与定位,探讨染料木黄酮(GEN)对新生小鼠睾丸组织PPP2R2C、细胞周期蛋白依赖性激酶2(CDK2)和波形蛋白(Vimentin)表达的影响。方法选用出生后1、6、10、15、20、30、60 d的雄性小鼠,采用免疫荧光技术观察PPP2R2C蛋白在各发育阶段小鼠睾丸中的细胞定位和表达;用2~3日龄雄性小鼠,取睾丸组织随机分为5组,即DMSO溶剂对照组和4个GEN剂量组(终浓度6.25、12.5、25、50μmol/L)。旋转通气法体外培养72 h,HE染色观察睾丸组织病理改变,Real-time PCR测定蛋白PPP2R2C、CDK2和Vimentin m RNA表达。结果免疫荧光结果发现PPP2R2C蛋白在小鼠睾丸间质细胞和其他细胞胞浆内表达;HE染色显示,各GEN剂量组均出现睾丸组织病理改变;Real-time PCR检测显示,6.25μmol/L GEN组PPP2R2C m RNA表达高于对照组(P=0.021),各剂量组CDK2 m RNA表达呈下降趋势(P>0.05),GEN各剂量组Vimentin m RNA表达略有降低,但是差异无统计学意义(P>0.05)。结论本研究首次发现PPP2R2C蛋白在小鼠睾丸组织中表达,其可能参与了小鼠睾丸发育、细胞分裂和精子发生过程。GEN可能通过调节体外培养小鼠睾丸PPP2R2C、CDK2和Vimentin基因的表达,影响睾丸发育和精子发生过程。更多还原

【Abstract】 Objective To detect the expression and location of PPP2 R2 C protein in mouse testes at different development stages, and to explore the effect of genistein on PPP2 R2 C, cyclin-dependent protein kinase 2(CDK2) and Vimentin expression in neonatal mouse testes. Methods The cellular location and expression of PPP2 R2 C protein at various developmental stages in post-neonatal 1 day(1 d), 6 d, 10 d, 15 d, 20 d, 30 d, 60 d male Kunming mice testes were detected by immunofluorescence technique.2 to 3-days-old neonatal Kunming mice testes were divided into 5 groups which were treated with DMSO as solvent control group and others treated with genistein in different concentrations(6.25, 12.5, 25, 50 μmol/L,respectively). All testes were cultured in vitro for 72 h by rotary aerobic culture method. The histomorphological changes in testicular tissues were observed by HE staining. the m RNA expression of PPP2 R2 C, CDK2 and Vimentin in cultured testes were detected by real-time PCR. Results Immunofluorescence staining showed that PPP2 R2 C protein was expressed in cytoplasm of mouse testis Leydig cells and other testicular cells. HE staining showed that various doses of genistein groups lead to histomorphological changes in the testes. Real-time PCR reveals that m RNA expression of PPP2 R2 C in 6.25 μmol/L genistein was higher than that of control group(P=0.021). All doses of genistein groups might decrease the m RNA levels of CDK2 and Vimentin, but the difference had no statistical significance(P>0.05). Conclusion This study finds for the first time that PPP2 R2 C protein are expressed in mouse testis. PPP2 R2 C expression might be correlated with mouse testis development,cell division and spermatogenesis. Genistien might affect testis development and spermato genesis via regulating PPP2 R2 C,CDK2 and Vimentin gene expression.更多还原