NFAT2 is implicated in corticosterone-induced rat Leydig cell apoptosis
NFAT2 is implicated in corticosterone-induced rat Leydig cell apoptosis
复制标题
NFAT2 参与皮质酮诱导的大鼠 Leydig 细胞凋亡
DOI：
10.1111/j.1745-7262.2007.00257.x
复制
发表时间：
2007-09-01
影响因子：
2.9
通讯作者：
Gao, Hui-Bao
中科院分区：
文献类型：
--
作者：
Chai, Wei-Ran;Wang, Qian;Gao, Hui-Bao
目的：探讨活化T细胞核因子（NFAT）的活化及其在皮质酮（CORT）诱导大鼠睾丸间质细胞凋亡中的作用。方法：采用免疫印迹法和免疫组化法检测大鼠睾丸间质细胞NFAT的表达。环孢菌素A（CsA）被用来评估潜在的参与NFAT在皮质醇诱导的Leydig细胞凋亡。用Fluo-3/AM监测皮质激素处理的Leydig细胞内Ca ~（2+）。将Leydig细胞与CORT或CORT + CsA孵育12 h后，用半定量Western印迹法测定细胞核和细胞质中NFAT 2的水平。通过观察NFAT 2过表达和CsA抑制NFAT 2活化对FasL表达和凋亡的影响，进一步评价NFAT 2在CORT诱导Leydig细胞凋亡中的作用。结果：我们发现NFAT 2是Leydig细胞中的主要亚型。CsA可阻断CORT诱导的Leydig细胞凋亡。CORT处理后Leydig细胞内Ca ~（2+）水平显著升高。CORT使核内NFAT 2水平升高，胞浆内NFAT 2水平降低。CsA可阻断CORT诱导的Leydig细胞NFAT 2核转位。NFAT 2过表达可增强皮质醇诱导的大鼠睾丸间质细胞凋亡和FasL表达，CsA可拮抗这两种作用。结论：NFAT 2在CORT诱导的Leydig细胞凋亡中被激活。NFAT 2过表达和抑制NFAT 2活化的作用表明，NFAT 2可能通过上调FasL在CORT诱导的Leydig细胞凋亡中发挥促凋亡作用。
Aim: To investigate the activation of the nuclear factor of activated T cells (NFAT) and its function in the corticoster-one (CORT)-induced apoptosis of rat Leydig cells. Methods: NFAT in rat Leydig cells was detected by Western blotting and immunohistochemical staining. Cyclosporin A (CsA) was used to evaluate potential involvement of NFAT in the CORT-induced apoptosis of Leydig cells. Intracellular Ca2+ was monitored in CORT-treated Leydig cells using Fluo-3/AM. After the Leydig cells were incubated with either CORT or CORT plus CsA for 12 h, the levels of NFAT2 in the nuclei and in the cytoplasm were measured by semi-quantitative Western blotting. The role of NFAT2 in CORT-induced Leydig cell apoptosis was further evaluated by observing the effects of NFAT2 overexpression and the inhibition of NFAT2 activation by CsA on FasL expression and apoptosis. Results: We found that NFAT2 was the predominant isoform in Leydig cells. CsA blocked the CORT-induced apoptosis of the Leydig cells. The intracellular Ca2+ level in the Leydig cells was significantly increased after the CORT treatment. The CORT increased the level of NFAT2 in the nuclei and decreased its level in the cytoplasm. CsA blocked the CORT-induced nuclear translocation of NFAT2 in the Leydig cells. Both CORT-induced apoptosis and FasL expression in the rat Leydig cells were enhanced by the overexpression of NFAT2 and antagonized by CsA. Conclusion: NFAT2 was activated in CORT-induced Leydig cell apoptosis. The effects of NFAT2 overexpression and the inhibition of NFAT2 activation suggest that NFAT2 may potentially play a pro-apoptotic role in CORT-induced Leydig cell apoptosis through the up-regulation of FasL.