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接骨木通过BMP-2/Smad/p38/JNK/Runx2信号通路促进MC3T3-E1细胞增殖和分化。

Sambucus Williamsii Hance Promotes MC3T3-E1 Cells Proliferation and Differentiation via BMP-2/Smad/p38/JNK/Runx2 Signaling Pathway.

作者信息

Yang Bingyou, Lin Xiaoying, Yang Chunli, Tan Jinyan, Li Wei, Kuang Haixue

机构信息

Key laboratory of Chinese Materia Medica, Ministry of Education, Heilongjiang University of Chinese Medicine, Haerbin, 150040, China.

Department of Obstetrics and Gynecology, National Key Laboratory of Gynecology, First Affiliated Hospital, Heilongjiang University of Chinese Medicine, Harbin, 150040, China.

出版信息

Phytother Res. 2015 Nov;29(11):1692-9. doi: 10.1002/ptr.5482. Epub 2015 Oct 12.

Abstract

The 50% ethanol elution fractions of root-bark of Sambucus Williamsii Hance (rbSWH) evaluated the effect of proliferation and differentiation on preosteoblast MC3T3-E1 cell, and the mechanism of actions. We found that rbSWH(30, 60, and 90 µg/mL) can enhance cell proliferation by MTT assay and promote alkaline phosphatase (ALP) and bone Gla protein (BGP) activities, type I collagen (Col-I) synthesis, and mineralization nodule formation in primary cultured osteoblasts. The results showed that rbSWH can increase mRNA levels of BMP-2 and Runx2 using real-time reverse transcription-quantitative polymerase chain reaction, whereas the BMP-2 antagonist Noggin attenuated the increase of ALP activity induced by rbSWH, indicating that BMP-2 expression was required for the action of rbSWH in osteoblastic. We also found that rbSWH can enhance the expressions of BMP-2, BMPRIB, BMPRII, phosphorylation of Smad, JNK and p38, and Runx2 proteins by western blotting. In addition, pretreatment of cells with p38 inhibitor (SB203580) or JNK inhibitor (SP600125) can antagonize the elevation of BMP-2 expression, ALP activity, and cell viability induced by rbSWH. Taken together, our results provided an evidence that rbSWH can promote MC3T3-E1 cell proliferation and differentiation via BMP-2/Smad/p38/JNK/Runx2 signaling pathway.

摘要

接骨木根皮50%乙醇洗脱组分(rbSWH)评估了其对前成骨细胞MC3T3-E1细胞增殖和分化的影响及其作用机制。我们发现,通过MTT法检测,rbSWH(30、60和90μg/mL)可增强细胞增殖,并促进原代培养成骨细胞中碱性磷酸酶(ALP)和骨钙蛋白(BGP)活性、I型胶原蛋白(Col-I)合成以及矿化结节形成。结果表明,通过实时逆转录定量聚合酶链反应,rbSWH可增加骨形态发生蛋白-2(BMP-2)和Runx2的mRNA水平,而BMP-2拮抗剂Noggin减弱了rbSWH诱导的ALP活性增加,表明BMP-2表达是rbSWH在成骨细胞中发挥作用所必需的。我们还发现,通过蛋白质印迹法,rbSWH可增强BMP-2、BMPRIB、BMPRII、Smad、JNK和p38的磷酸化以及Runx2蛋白的表达。此外,用p38抑制剂(SB203580)或JNK抑制剂(SP600125)预处理细胞可拮抗rbSWH诱导的BMP-2表达升高、ALP活性增加和细胞活力增强。综上所述,我们的结果证明rbSWH可通过BMP-2/Smad/p38/JNK/Runx2信号通路促进MC3T3-E1细胞增殖和分化。

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