Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; and Department of Obstetrics and Gynaecological Ultrasound Imaging, Renmin Hospital of Wuhan University, No. 99 Zhang Zhidong Road, No. 238 Jiefang Road, Wuhan 430060, China.
Reprod Fertil Dev. 2020 Dec;32(18):1357-1364. doi: 10.1071/RD20253.
In this study, the ability of cold-induced RNA-binding protein (CIRBP) to regulate the expression of Src-associated during mitosis of 68 kDa (Sam68) and extracellular signal-regulated kinases (ERK) in the mouse testis and mouse primary spermatocytes (GC-2spd cell line) before and after heat stress was examined to explore the molecular mechanism by which CIRBP decreases testicular injury. A mouse testicular hyperthermia model, a mouse primary spermatocyte hyperthermia model and a low CIRBP gene-expression cell model were constructed and their relevant parameters were analysed. The mRNA and protein levels of CIRBP and Sam68 were significantly decreased in the 3-h and 12-h testicular heat-stress groups, extracellular signal-regulated kinase 1/2 (ERK1/2) protein expression was not significantly affected but phospho-ERK1/2 protein levels were significantly decreased. GC-2spd cellular heat-stress results showed that the mRNA and protein concentrations of CIRBP and Sam68 were reduced 48h after heat stress. In the low CIRBP gene-expression cell model, CIRBP protein expression was significantly decreased. Sam68 mRNA expression was significantly decreased only at the maximum transfection concentration of 50nM and Sam68 protein expression was not significantly affected. These findings suggest that CIRBP may regulate the expression of Sam68 at the transcriptional level and the expression of phospho-ERK1/2 protein, both of which protect against heat-stress-induced testicular injury in mice.
在这项研究中，研究人员探讨了冷诱导 RNA 结合蛋白 (CIRBP) 在热应激前后调节小鼠睾丸和小鼠初级精母细胞（GC-2spd 细胞系）中 Src 相关在有丝分裂中 68 kDa (Sam68) 和细胞外信号调节激酶 (ERK) 的表达的能力，以探索 CIRBP 降低睾丸损伤的分子机制。构建了小鼠睾丸过热模型、小鼠初级精母细胞过热模型和 CIRBP 低表达基因细胞模型，并分析了它们的相关参数。在 3 小时和 12 小时睾丸热应激组中，CIRBP 和 Sam68 的 mRNA 和蛋白水平显著降低，细胞外信号调节激酶 1/2 (ERK1/2) 蛋白表达无明显变化，但磷酸化 ERK1/2 蛋白水平显著降低。GC-2spd 细胞热应激结果显示，热应激 48 小时后，CIRBP 和 Sam68 的 mRNA 和蛋白浓度降低。在低 CIRBP 基因表达细胞模型中，CIRBP 蛋白表达显著降低。仅在最大转染浓度 50nM 时，Sam68 mRNA 表达显著降低，而 Sam68 蛋白表达无明显影响。这些发现表明，CIRBP 可能通过调节转录水平的 Sam68 表达和磷酸化 ERK1/2 蛋白表达，来保护小鼠免受热应激引起的睾丸损伤。
