受激活调节正常T细胞表达和分泌因子及其受体CCR1、CCR5在大鼠附睾的表达与定位

摘要

Objective:To observe the expression and the cellular localization of regulated upon activation normal T-cell expressed and secreted (RANTES) as well as its receptors CCR1 and CCR5 in the rat epididymis.Methods:Reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of RANTES and its receptors mRNA in rat epididymis.The presence of RANTES and its receptors protein was assayed by Western blotting.Paraffin-embedded sections were prepared from rat epididymis,and immunohistochemical staining was carried out to detect cellular location of RANTES and its receptors.Colocalization of RANTES,CCR1,and CCR5 in rat epididymis was detected by immunofluorescence.Results:RT-PCR analysis showed the specific expression of RANTES and its receptors in the segments of adult rat epididymis.The distinctive single band of RANTES protein and its receptors protein was identified in the segments of rat epididymis with Western blotting.The positive staining of RANTES and its receptors were mainly observed in the basal cells of the epididymal epithelium.The immunofluorescence of RANTES coincided with CCR1 and CCR5 in the basal cells of epididymis.Conclusion:The expressions of RANTES and its receptors CCR1 and CCR5 were found restricted in the basal cells of the rat epididymis.These results revealed that RANTES may play a role in the physiological function of the basal cells of rat epididymis through autocrine or/and paracrine way.%目的:观察受激活调节正常T细胞表达和分泌因子(RANTES)与其受体CCR1、CCR5在大鼠附睾中的表达和定位.方法:采用免疫组织化学法检测成年大鼠附睾上皮RANTES及其受体的细胞定位,免疫荧光双标染色分别显示RANTES与CCR1、CCR5的细胞共定位情况.RT-PCR检测RANTES及其受体mRNA表达水平,免疫印迹检测RANTES及其受体的蛋白量.结果:RANTES与其受体CCR1、CCR5在大鼠附睾各段呈特异性表达.免疫印迹在大鼠附睾各段检测到RANTES及其受体CCR1、CCR5特异性蛋白条带.RANTES与其受体CCR1、CCR5在大鼠附睾上皮主要表达于基细胞.双重免疫荧光显示RANTES与CCR1、CCR5在附睾管基细胞共存.结论:RANTES可能通过自分泌或(和)旁分泌方式在大鼠附睾基细胞功能活动方面起重要作用.