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观察锌指蛋白265(ZNF265)基因对睾丸Sertoli细胞溶脲脲原体(Ureaplasma urealyticum,UU)感染后IL-1α、IL-6及TGF-β1表达的影响,并探讨其可能的作用机制。构建沉默大鼠ZNF265基因表达的慢病毒载体,体外转染Sertoli细胞,对转染阳性的细胞进行UU感染,在感染后的第1、3、5天分别检测对照组和实验组中IL-1α、IL-6及TGF-β1表达。结果显示,成功构建干扰Sertoli细胞中ZNF265表达的慢病毒载体,在Sertoli细胞中转染效率高达62.8%,在mRNA水平和蛋白质水平干扰ZNF265表达的效率分别为65.73%和94%;干扰Sertoli细胞中ZNF265表达后,在UU感染的第1天和第3天,IL-1α、IL-6及TGF-β1表达升高;在UU感染第5天,ZNF265表达较对照组升高,TGF-β1表达升高,但IL-1α,IL-6表达下降。以上结果说明,构建的慢病毒载体成功地干扰ZNF265基因在大鼠Sertoli细胞中的表达,干扰其表达后,IL-1α和IL-6表达增加;ZNF265作为调控因子,可能通过作用于TGF-β1,在抑制炎症发展、维持睾丸局部免疫微环境的平衡中发挥重要作用。
To investigate the effect of zinc finger protein 265 (ZNF265) on the expression of IL-1α, IL-6 and TGF-β1 in Sertoli cells after Ureaplasma urealyticum infection and to explore its possible mechanism. To construct the lentiviral vector expressing silenced rat ZNF265 gene, transfect Sertoli cells in vitro and UU infection in transfected cells. The levels of IL-1α in control group and experimental group were detected on the 1st, 3rd, 5th day after infection , IL-6 and TGF-β1 expression. The results showed that the lentiviral vector that interfered with the expression of ZNF265 in Sertoli cells was successfully constructed. The transfection efficiency of Sertoli cells was up to 62.8%, and the efficiency of interfering ZNF265 expression at mRNA and protein level was 65.73% and 94%, respectively. After the expression of ZNF265, the expression of IL-1α, IL-6 and TGF-β1 increased on the first day and the third day of UU infection. On the fifth day after UU infection, the expression of ZNF265 was increased, the expression of TGF-β1 Increased, but IL-1α, IL-6 expression decreased. The above results indicated that the constructed lentiviral vector successfully interfered with the expression of ZNF265 gene in Sertoli cells, and the expression of IL-1α and IL-6 was increased after interfering with the expression of ZNF265 gene. ZNF265 could regulate the expression of TGF-β1 , Play an important role in suppressing the development of inflammation and maintaining the balance of local immune microenvironment in the testis.