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首页> 外文期刊>Biochemistry >1α,25-Dihydroxyvitamin D_3 Signaling Pathways on Calcium Uptake in 30-Day-Old Rat Sertoli Cells
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1α,25-Dihydroxyvitamin D_3 Signaling Pathways on Calcium Uptake in 30-Day-Old Rat Sertoli Cells

机译：1α，25-二羟基维生素D_3信号通路对30天大的大鼠支持细胞中钙摄取的影响

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1α,25-Dihydroxyvitamin D3_ (1,25D_3) is the active metabolite of vitamin D_3 and the major calcium regulatory hormone in tissues. The aim of this work was to investigate the mechanism of action of 1,25D_3 on 45Ca~(2+) uptake in Sertoli cells from 30-day-old rats. Results showed that 10~(-9) and 10~(-12) M 1,25D_3 increased the rate of 45Ca~(2+) uptake 5 and 15 min after hormone exposure and that 1α,25(OH)2 lumisterol3 (JN) produced a similar effect suggesting that 1,25D_3 action occurs via a putative membrane receptor. The involvement of voltage-dependent calcium channels (VDCC) in 1,25D_3 action was evidenced by using nifedipine, while the use of Bapta-AM demonstrated that intracellular calcium was not implicated. Moreover, the incubation with ouabain and digoxin increased the rate of ~(45)Ca~(2+) uptake, indicating that the effect of 1,25D_3 may also result from Na~+/K~+-ATPase inhibition. In addition, we demonstrated that the mechanism underlying the hormone action involved extracellular signal-regulated kinase (ERK) and protein kinase C (PKC) activation in a phospholipase C-independent way. Furthermore, a local elevation of the level of cAMP, as demonstrated by incubating cells with dibutyryl cAMP or a phosphodiesterase inhibitor, produced an effect similar to that of 1,25D_3, and the inhibition of protein kinase A (PKA) nullified the hormone action. In conclusion, the stimulatory effect of 1,25D_3 on 45Ca~(2+) uptake in Sertoli cells occurs via VDCC, as well as PKA, PKC, and ERK activation. These protein kinases seem to act by inhibiting Na~+/K~+-ATPase or directly osphorylating calcium channels. The Na~+/K~+-ATPase inhibition may result in Na~+/Ca~(2+) exchanger activation in reverse mode and consequently induce the uptake of calcium into the cells.
机译：1α，25-二羟基维生素D3_（1,25D_3）是维生素D_3的活性代谢产物，是组织中的主要钙调节激素。这项工作的目的是研究1,25D_3对30日龄大鼠Sertoli细胞45Ca〜（2+）摄取的作用机制。结果显示，在激素暴露后5和15分钟，10〜（-9）和10〜（-12）M 1,25D_3增加45Ca〜（2+）的摄取速率，而1α，25（OH）2 lumisterol3（JN ）产生类似的作用，表明1,25D_3的作用是通过推定的膜受体发生的。硝苯地平证明了电压依赖性钙通道（VDCC）参与1,25D_3的作用，而Bapta-AM的使用表明不涉及细胞内钙。此外，与哇巴因和地高辛的孵育增加了〜（45）Ca〜（2+）的吸收速率，表明1,25D_3的作用也可能是由Na〜+ / K〜+ -ATPase抑制引起的。此外，我们证明了激素作用的潜在机制涉及以磷脂酶C独立的方式参与细胞外信号调节激酶（ERK）和蛋白激酶C（PKC）的激活。此外，通过将细胞与二丁酰基cAMP或磷酸二酯酶抑制剂一起孵育来证明，cAMP水平的局部升高产生了类似于1,25D_3的作用，并且蛋白激酶A（PKA）的抑制使激素作用无效。总之，1,2,5D_3对Sertoli细胞45Ca〜（2+）摄取的刺激作用是通过VDCC以及PKA，PKC和ERK激活产生的。这些蛋白激酶似乎是通过抑制Na〜+ / K〜+ -ATPase或直接磷酸化钙通道而起作用。 Na〜+ / K〜+ -ATPase的抑制作用可能导致Na〜+ / Ca〜（2+）交换子以相反的方式活化，从而诱导钙被细胞吸收。

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