Testicular tissue expression of SIRT1 gene in Qianbei Ma goat and effects of its overexpression on Leydig cell development and antioxidant capacity

【Objective】The expression levels of silent regulator 1(SIRT1) in testicular tissues of different months old and gonadal axis tissues of 12 months old Qianbei Ma goat were analyzed,and the effects of SIRT1 on the development and antioxidant capacity of testis Leydig cells were analyzed,which provided a theoretical basis for exploring the mechanism of SIRT1 gene regulating the growth and development of testis Leydig cells. 【Method】The expression of SIRT1 in testis and gonadal axis(hypothalamus,pituitary,testis and epididymis) was detected by real-time fluorescence quantitative PCR in healthy Qianbei Ma goat rams at 1, 3, 6, 9 and 12 months old. The overexpression vector pEGFP-N3-SIRT1and the empty vector pEGFP-N3 were transfected into goat testis Leydig cells. CCK-8 method was used to detect cell proliferation. Scratch test was used to detect cell migration efficiency. Real-time fluorescence quantitative PCR was used to detect the expression of cell proliferation,cell development and antioxidant related genes. 【Result】The expression level of SIRT1 gene in testicular tissue of 3, 6 and 9 months old was significantly higher than that of 1 month old(P<0.05, the same below),and that of 12 months old was the highest. On the gonadal axis,the relative expression of SIRT1 gene in testicular tissue was significantly higher than that in hypothalamus,pituitary and epididymis. The results of CCK-8 method showed that overexpression of SIRT1 gene extremely significantly promoted the proliferation of testis Leydig cells after 12 h(P<0.01). Scratch test showed that the cell mobility of overexpression SIRT1 gene group was significantly higher than that of no-load group at 9 and 18 h. Real-time fluorescence quantitative PCR results showed that overexpression of SIRT1gene could significantly promote testicular development related gene insulin-like growth factor 2(IGF2), as well as expression of cell proliferation-related genes proliferating cell nuclear antigen(PCNA) and antioxidant-related genes superoxide dismutase(SOD), glutathione peroxidase 1(GPx1) and heme oxygenase-1(HO-1), the expression of nuclear factor E2 related factor 2(Nrf2) was not significantly effected(P>0.05). 【Conclusion】SIRT1 gene is expressed in testicular tissue in different months of Qianbei Ma goat. The overexpressed SIRT1 gene is successfully expressed in testis Leydig cells, and it may promote cell proliferation and development and improve its antioxidant capacity by increasing the expression of related genes in Qianbei Ma goat testis Leydig cells, thus directly or indirectly affecting the physiological function of testis Leydig cells.