Engineering Laboratory of Animal Pharmaceuticals, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian Province, 350002, PR China; Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China; Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, 12100, China.
Engineering Laboratory of Animal Pharmaceuticals, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian Province, 350002, PR China.
Theriogenology. 2021 Feb;161:140-150. doi: 10.1016/j.theriogenology.2020.11.010. Epub 2020 Dec 3.
Luman has been reported to be involved in the formation of COP II-mediated transport vesicles that affect protein transportation and secretion. Western blotting, immunohistochemistry, immunofluorescence, and RT-qPCR indicated that Luman is widely expressed in the male mouse reproductive system. In sperm, Luman was mainly located in the sperm tail, and the expression level increased with sperm maturity. In the testis, Luman was located in Leydig cells. In MLTC-1, a high-concentration hCG treatment significantly increased GRP78, ATF6, p-IRE1, and p-EIF2S1 expression but had no effect on Luman expression. To investigate the role of Luman in hCG-induced ER stress (ERS), experiments were conducted to examine the consequences of short hairpin RNA (shRNA)-mediated Luman knockdown in MLTC-1 cells. Luman knockdown decreased the percentage of S phase cells and up-regulated Cyclin A1, Cyclin B1, and Cyclin D2 expression. ELISA and WB results showed that with Luman knockdown, Cyp11a1, p-IRE1, and p-EIF2S1 expression and testosterone secretion were significantly increased, while GRP78 and CHOP expression were decreased. Flow cytometry results showed that Luman knockdown reduced MLTC-1 cell apoptosis. RT-qPCR and WB results showed that Luman knockdown significantly up-regulated BCL-2 expression and decreased Caspase-3 and BAX expression. These data suggest that Luman is widely expressed in the male mouse reproductive system. In MLTC-1 cells, Luman knockdown up-regulated p-IRE1, p-EIF2S1, and BCL-2 expression and decreased GRP78, CHOP, BAX, and Caspase-3 expression. We propose that Luman knockdown reduces cell apoptosis through the ERS pathway, thereby promoting cell survival and testosterone secretion. These findings provide new insights into the role of Luman in hCG-induced ERS.
鲁曼（Luman）被报道参与了 COP II 介导的运输小泡的形成，该小泡影响蛋白质的运输和分泌。Western blot、免疫组织化学、免疫荧光和 RT-qPCR 表明，Luman 在雄性小鼠生殖系统中广泛表达。在精子中，Luman 主要位于精子尾部，其表达水平随精子成熟而增加。在睾丸中，Luman 位于间质细胞中。在 MLTC-1 细胞中，高浓度 hCG 处理显著增加了 GRP78、ATF6、p-IRE1 和 p-EIF2S1 的表达，但对 Luman 的表达没有影响。为了研究 Luman 在 hCG 诱导的内质网应激（ERS）中的作用，进行了实验以研究短发夹 RNA（shRNA）介导的 Luman 敲低对 MLTC-1 细胞的影响。Luman 敲低降低了 S 期细胞的百分比，并上调了 Cyclin A1、Cyclin B1 和 Cyclin D2 的表达。ELISA 和 WB 结果表明，随着 Luman 的敲低，Cyp11a1、p-IRE1 和 p-EIF2S1 的表达和睾酮分泌显著增加，而 GRP78 和 CHOP 的表达降低。流式细胞术结果表明，Luman 敲低减少了 MLTC-1 细胞的凋亡。RT-qPCR 和 WB 结果表明，Luman 敲低显著上调了 BCL-2 的表达，降低了 Caspase-3 和 BAX 的表达。这些数据表明，Luman 在雄性小鼠生殖系统中广泛表达。在 MLTC-1 细胞中，Luman 敲低上调了 p-IRE1、p-EIF2S1 和 BCL-2 的表达，降低了 GRP78、CHOP、BAX 和 Caspase-3 的表达。我们提出，Luman 敲低通过 ERS 途径减少细胞凋亡，从而促进细胞存活和睾酮分泌。这些发现为 Luman 在 hCG 诱导的 ERS 中的作用提供了新的见解。
