Department of Reproductive Medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, 201204, China.
Laboratory for Reproductive Immunology, Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical College, Shanghai, 200011, China.
Reprod Biol Endocrinol. 2020 Oct 12;18(1):100. doi: 10.1186/s12958-020-00658-0.
Trophoblast cells are required for the establishment of pregnancy and fetal development. Apoptosis is an essential feature for trophoblast invasion. Uncontrolled trophoblast apoptosis is related to some complicate pregnancies. Oxidative stress (OS) is an important inducer of trophoblast apoptosis. Cyclosporin A (CsA) has been shown to promote the activity of trophoblast cells and reduce OS-induced oxidative injury. We investigated the role and mechanism of CsA in oxidative stress-induced trophoblast cell apoptosis.
JEG-3 cells were cocultured with HO and CsA. Cell viability and morphology were measured by MTT assay and DAPI staining. Cell apoptosis was tested with annexin V/PI staining. The expression of Bcl-2-associated X protein (Bax), B-cell lymphoma/leukemia-2 (Bcl-2), cleaved poly (ADP-ribose) polymerase (PARP) and pro-caspase-3 was assayed by western blotting. The protein expression and phosphorylation of p53 and mitogen-activated protein kinase (MAPK) kinases (JNK, ERK1/2 and p38) were examined by western blotting.
CsA increased the viability, alleviated morphological injury and reduced cell apoptosis of the HO-treated JEG-3 cells. CsA also attenuated the activation of p53, decreased the expression of Bax and cleavage of PARP, and increased the expression of Bcl-2 and pro-caspase-3 in the JEG-3 treated with HO. Furthermore, CsA reduced the activation of JNK and P38 but had no significant effect on the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the HO-treated JEG-3 cells. Promoting the activation of JNK and p38 impaired the protective effect of CsA on OS-induced trophoblast apoptosis.
These results suggested that CsA protected trophoblast cells from OS-induced apoptosis via the inhibition of the p53 and JNK/p38 signaling pathways.
滋养细胞是妊娠和胎儿发育所必需的。细胞凋亡是滋养细胞侵袭的一个重要特征。滋养细胞凋亡失控与一些复杂的妊娠有关。氧化应激（OS）是诱导滋养细胞凋亡的重要因素。环孢素 A（CsA）已被证明能促进滋养细胞的活性并减少 OS 诱导的氧化损伤。我们研究了 CsA 在氧化应激诱导的滋养细胞凋亡中的作用和机制。
JEG-3 细胞与 HO 和 CsA 共培养。用 MTT 法和 DAPI 染色检测细胞活力和形态。用 annexin V/PI 染色检测细胞凋亡。用 Western blot 检测 Bcl-2 相关 X 蛋白（Bax）、B 细胞淋巴瘤/白血病-2（Bcl-2）、多聚（ADP-核糖）聚合酶（PARP）和前半胱天冬酶-3 的表达。用 Western blot 检测 p53 和丝裂原活化蛋白激酶（MAPK）激酶（JNK、ERK1/2 和 p38）的蛋白表达和磷酸化。
CsA 增加了 HO 处理的 JEG-3 细胞的活力，减轻了形态损伤，减少了细胞凋亡。CsA 还减弱了 p53 的激活，降低了 Bax 的表达和 PARP 的切割，增加了 HO 处理的 JEG-3 细胞中 Bcl-2 和前半胱天冬酶-3 的表达。此外，CsA 降低了 JNK 和 P38 的激活，但对 HO 处理的 JEG-3 细胞中细胞外信号调节激酶 1/2（ERK1/2）的激活没有显著影响。促进 JNK 和 p38 的激活会损害 CsA 对 OS 诱导的滋养细胞凋亡的保护作用。
这些结果表明，CsA 通过抑制 p53 和 JNK/p38 信号通路来保护滋养细胞免受 OS 诱导的凋亡。
