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雌二醇增强大鼠骨髓基质细胞成骨作用与 JNK 通路有关。

Estradiol‑enhanced osteogenesis of rat bone marrow stromal cells is associated with the JNK pathway.

机构信息

Key Laboratory of Ministry of Education for TCM Viscera‑State Theory and Applications, Ministry of Education of China, Shenyang, Liaoning 110847, P.R. China.

The Graduate School, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China.

出版信息

Mol Med Rep. 2017 Dec;16(6):8589-8594. doi: 10.3892/mmr.2017.7699. Epub 2017 Oct 3.

Abstract

Bone marrow stromal cells (BMSCs) can differentiate into osteoblasts. The present study investigated the osteogenic effects of estradiol, as well as the role of the c‑Jun N‑terminal kinase (JNK) signaling pathway in promoting estradiol‑enhanced osteogenesis of rat (r)BMSCs. rBMSCs were treated for 7 days with or without estradiol and further treated with or without the JNK‑specific inhibitor SP600125. The role of estrogen during rBMSC osteogenesis was evaluated by alkaline phosphatase activity and mineralized nodule formation using the Gomori method and Alizarin red S staining, respectively. Subsequently, the mRNA expression levels of transforming growth factor-β1 (TGF‑β1) and core‑binding factor α1 (Cbfα1) were evaluated by reverse transcription‑quantitative polymerase chain reaction, and TGF‑β1, Cbfα1 and phosphorylated (p)‑JNK protein expression was detected by western blotting. All groups treated with SP600125 expressed low levels of TGF‑β1 and Cbfα1 mRNA and protein, and low p‑JNK protein expression. Compared with the control cells, rBMSCs cultured with estradiol exhibited a significant upregulation in the expression levels of osteogenic genes and proteins. The present study demonstrated that estradiol enhanced osteogenic differentiation of rBMSCs and that the JNK signaling pathway was involved in this process, providing insights into the molecular mechanisms involved in rBMSC osteogenesis upon estradiol stimulation.

摘要

骨髓基质细胞(BMSCs)可分化为成骨细胞。本研究探讨了雌二醇的成骨作用,以及 c-Jun N-末端激酶(JNK)信号通路在促进大鼠(r)BMSCs 中雌二醇增强成骨作用中的作用。用或不用雌二醇处理 rBMSCs7 天,并进一步用或不用 JNK 特异性抑制剂 SP600125 处理。通过碱性磷酸酶活性和用 Gomori 法和茜素红 S 染色分别形成矿化结节来评估雌激素在 rBMSC 成骨过程中的作用。随后,通过逆转录-定量聚合酶链反应评估转化生长因子-β1(TGF-β1)和核心结合因子α1(Cbfα1)的 mRNA 表达水平,并通过 Western blot 检测 TGF-β1、Cbfα1 和磷酸化(p)-JNK 蛋白表达。所有用 SP600125 处理的组均表达低水平的 TGF-β1 和 Cbfα1 mRNA 和蛋白,以及低水平的 p-JNK 蛋白表达。与对照细胞相比,用雌二醇培养的 rBMSCs 中成骨基因和蛋白的表达水平显著上调。本研究表明,雌二醇增强了 rBMSCs 的成骨分化,而 JNK 信号通路参与了这一过程,为雌二醇刺激 rBMSC 成骨过程中的分子机制提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cfe/5779911/1cbf1a24010f/MMR-16-06-8589-g00.jpg

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