胰岛素抵抗状态下滋养细胞中钙/钙调蛋白依赖性蛋白激酶 IV：功能与代谢组学分析。
Calcium/calmodulin dependent protein kinase IV in trophoblast cells under insulin resistance: functional and metabolomic analyses.
机构信息
Department of Endocrinology, Shengjing Hospital of China Medical University, 36 Sanhao Street, Shenyang, 110004, Liaoning, People's Republic of China.
出版信息
Mol Med. 2023 Jun 29;29(1):82. doi: 10.1186/s10020-023-00669-8.
BACKGROUND
Insulin resistance (IR) is an important determinant of glucose metabolic disturbance and placental dysplasia in gestational diabetes mellitus (GDM). Calcium/calmodulin dependent protein kinase IV (CAMK4) improves insulin IR induced by a high-fat diet (HFD). The current study sought to elucidate the role and potential mechanism of CAMK4 in GDM.
METHODS
A GDM model was established in female C57BL/6J mice via HFD feeding for one week before mating and throughout gestation. The IR was elicited by 10 M insulin treatment for 48 h in HTR-8/SVneo cells and mouse primary trophoblast cells. The function of CAMK4 was investigated by transfection of overexpression plasmid in HTR-8/SVneo cells and infection of lentivirus loaded with CAMK4 encoding sequence in primary trophoblast cells. Real-time PCR, western blot, cell counting kit-8, transwell, wound healing, dual-luciferase reporter assay, and liquid chromatography/mass spectrometry-based untargeted metabolomics were performed to confirm the effects of CAMK4 on trophoblast cells.
RESULTS
Decreased CAMK4 expression was found in the placenta of GDM mice. CAMK4 overexpression ameliorated IR-induced viability impairment, migratory and invasive capacity inhibition, autophagy blocking, insulin signaling inactivation and glucose uptake disorder in trophoblast cells. CAMK4 also transcriptionally activated orphan nuclear receptor NUR77, and the effects of CAMK4 were abrogated by silencing of NUR77. Metabolomics analysis revealed that CAMK4 overexpression caused alterations of amino acid, lipid and carbohydrate metabolism, which were important in GDM.
CONCLUSION
Our results indicated that CAMK4/NUR77 axis may provide novel potential targets in GDM treatment.
背景
胰岛素抵抗（IR）是妊娠期糖尿病（GDM）中葡萄糖代谢紊乱和胎盘发育不良的重要决定因素。钙/钙调蛋白依赖性蛋白激酶 IV（CAMK4）可改善高脂肪饮食（HFD）引起的胰岛素 IR。本研究旨在阐明 CAMK4 在 GDM 中的作用及潜在机制。
方法
通过在交配前一周和整个妊娠期用 HFD 喂养雌性 C57BL/6J 小鼠建立 GDM 模型。在 HTR-8/SVneo 细胞和小鼠原代滋养层细胞中，用 10 μM 胰岛素处理 48 h 诱导 IR。通过转染过表达质粒在 HTR-8/SVneo 细胞中研究 CAMK4 的功能，通过感染携带 CAMK4 编码序列的慢病毒在原代滋养层细胞中研究 CAMK4 的功能。通过实时 PCR、western blot、细胞计数试剂盒-8、transwell、划痕愈合、双荧光素酶报告基因检测和基于液相色谱/质谱的非靶向代谢组学，证实了 CAMK4 对滋养层细胞的作用。
结果
在 GDM 小鼠的胎盘组织中发现 CAMK4 表达降低。CAMK4 过表达改善了 IR 诱导的滋养层细胞活力损伤、迁移和侵袭能力抑制、自噬阻断、胰岛素信号失活和葡萄糖摄取障碍。CAMK4 还转录激活孤儿核受体 NUR77，沉默 NUR77 可阻断 CAMK4 的作用。代谢组学分析表明，CAMK4 过表达导致氨基酸、脂质和碳水化合物代谢的改变，这对 GDM 很重要。
结论
我们的研究结果表明，CAMK4/NUR77 轴可能为 GDM 的治疗提供新的潜在靶点。
Zotero 插件