旨在探究抑制水牛BET(bromodomain and extra terminal,BET)蛋白对支持细胞自噬调控的影响.以水牛支持细胞(sertoli cells,SCs)为研究对象,首先从睾丸中分离纯化得到高纯度的SCs,通过免疫荧光法及RT-PCR法检测出水牛SCs的标记基因均正常表达.结果显示,BET蛋白被抑制后,水牛SCs形态和数量发生显著变化,细胞的体积变大,数量减少,折光性减弱,免疫荧光分析和MDC分析也证实其生物学功能受到影响.超微结构分析发现,水牛SCs中产生大量自噬体、自噬溶酶体以及自噬空泡,且自噬相关标记基因的表达量显著提升.说明抑制BET蛋白可引起水牛SCs活性和自噬活性降低,并导致自噬体增多.结果表明,本试验为BET蛋白调控雄性生殖机制的研究提供了依据.
Effect of inhibiting BET protein on autophagy in buffalo sertoli cells
The aim of this study was to investigate the autophagy in buffalo sertoli cells with BET(bromodomain extra terminal)protein inhibition.Firstly,buffalo sertoli cells(SCs)were isolated and purified from testis.The marker genes of SCs were detected by immunofluorescence and RT-PCR.The BET protein inhibition of buffalo SCs resulted in significant changes in their morphology and number.The cells increased in size,decreased in number and refraction.Immunofluorescence and MDC analysis also confirmed that the biological functions of SCs were affected.Ultrastructural analysis showed that a large number of autophagosomes,autophagic lysosomes and autophagic ves-icles were produced in buffalo SCs.The expression level of marker genes associate with autophagy was significantly increased.These results indicated that inhibition of BET protein could decrease cell viability and autophagy in buffalo SCs,and lead to the increase of autophagosomes.This study provides basis for the research on the mechanism of BET protein regulation in male reproduction.
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