Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway.
Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway.
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DOI：
10.4103/1008-682x.160260
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发表时间：
2016-05
影响因子：
2.9
通讯作者：
Yao B
中科院分区：
文献类型：
--
作者：
He Z;Sun Q;Liang YJ;Chen L;Ge YF;Yun SF;Yao B
本研究旨在探讨膜联蛋白A5对原代大鼠睾丸间质细胞睾酮分泌的影响及其机制。用膜联蛋白A5处理分离的大鼠Leydig细胞。用化学发光法检测睾酮的产生。采用Western blotting和半定量RT-PCR方法分别检测星星、P450 SCC、3β-羟基类固醇脱氢酶（3β-HSD）、17β-羟基类固醇脱氢酶（17β-HSD）和17α-羟化酶（17α-hydroxylase）的蛋白和mRNA水平。Annexin A5以剂量和时间依赖性方式显著刺激大鼠Leydig细胞分泌睾酮，并增加星星、P450 SCC、3β-HSD和17β-HSD的mRNA和蛋白表达，但不增加17α-羟化酶的表达。Annexin A5 siRNA敲除可显著降低睾酮水平和P450 SCC、3β-HSD和17β-HSD蛋白表达。在膜联蛋白A5处理后5、10和30 min观察到ERK 1/2信号的显著激活。用ERK抑制剂PD 98059（50 μmol l−1）预处理Leydig细胞20 min后，Annexin A5促进睾酮分泌及P450 SCC、3β-HSD和17β-HSD表达的作用完全消失（P < 0.05）。因此，ERK 1/2信号转导参与了膜联蛋白A5在介导睾丸间质细胞中睾酮产生和P450 SCC、3β-HSD和17β-HSD表达中的作用。
This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3β-HSD, and 17β-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3β-HSD, and 17β-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 μmol l−1) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3β-HSD, and 17β-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3β-HSD, and 17β-HSD in Leydig cells.