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17β-雌二醇对犬骨髓间充质干细胞的体外增殖和凋亡有影响。

17 beta-estradiol affects proliferation and apoptosis of canine bone marrow mesenchymal stem cells in vitro.

作者信息

Zhou Z-H, Gu C-W, Li J, Huang X-Y, Deng J-Q, Shen L-H, Cao S-Z, Deng J-L, Zuo Z-C, Wang Y, Ma X-P, Ren Z-H, Yu S-M

机构信息

College of Veterinary Medicine, Sichuan Agricultural University, 211 Huimin Road, Chengdu 611130, China.

Laboratory Animal Centre, Southwest Medical University, 1 Xianglin Road, Luzhou 646000, China.

出版信息

Pol J Vet Sci. 2020 Jun;23(2):235-245. doi: 10.24425/pjvs.2020.133638.

Abstract

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apop-tosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin--dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluores-cence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly,cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

摘要

在人类、猪和小鼠中的新兴研究表明,雌激素在骨髓间充质干细胞(BMSCs)的自我更新和分化中起关键作用。本研究旨在评估17β-雌二醇(E2)对犬源骨髓间充质干细胞(cBMSCs)体外增殖和凋亡的影响。结果表明,通过CCK-8检测和对增殖相关基因的RT-qPCR分析,10-11 M浓度的E2补充促进了cBMSCs的增殖,增殖细胞核抗原(PCNA)、细胞周期蛋白D1(CCND1)上调,细胞周期蛋白依赖性激酶抑制剂1B(CDKN1B)下调。相反,荧光激活细胞分选(FACS)分析和RT-qPCR表明,高于10-11 M的E2补充对cBMSCs的增殖有抑制作用并诱导凋亡。有趣的是,添加10-11 M E2时,cBMSCs仍具有分化为成骨细胞和脂肪细胞的能力。综上所述,本研究确定了cBMSCs体外的最佳培养条件,对进一步了解E2对cBMSCs自我更新的调节作用具有重要意义,这有助于BMSCs的临床应用。

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