[1]李 洋,陈前昭,邵 英,等.骨形态蛋白9诱导干细胞骨向分化与环氧酶-2及PI3K/Akt的关系研究[J].中国药理学通报,2017,(07):908-915.[doi:10.3969/j.issn.1001-1978.2017.07.006]
 LI Yang,CHEN Qian-Zhao,SHAO Ying,et al.Study on relationship between BMP9-induced osteogenic differentiation and COX-2/PI3K/Akt in stem cells[J].Chinese Pharmacological Bulletin,2017,(07):908-915.[doi:10.3969/j.issn.1001-1978.2017.07.006]
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骨形态蛋白9诱导干细胞骨向分化与环氧酶-2及PI3K/Akt的关系研究()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2017年07期
页码:
908-915
栏目:
论著
出版日期:
2017-07-10

文章信息/Info

Title:
Study on relationship between BMP9-induced osteogenic differentiation and COX-2/PI3K/Akt in stem cells
文章编号:
1001-1978(2017)07-0908-08
作者:
李 洋12陈前昭12邵 英12曾于桦12任文艳12刘荣兴12何百成12
1. 重庆医科大学药学院,2. 重庆市生物化学与分子药理学重点实验室,重庆 400016
Author(s):
LI Yang12 CHEN Qian-Zhao12 SHAO Ying12ZENG Yu-hua12 REN Wen-yan12LIU Rong-xing12 HE Bai-cheng12
1.School of Pharmacy, Chongqing Medical University, Chongqing 400016,China;
2.Chongqing Key Lab of Biochemistry and Molecular Pharmacology, Chongqing 400016, China
关键词:
骨形态蛋白9 干细胞 成骨分化 成骨标志物 PI3K/Akt 环氧酶-2
Keywords:
bone morphogenetic protein 9 stem cell osteogenic differentiation osteogenic marker PI3K/Akt cycloxygenase 2
分类号:
R341;R329.2;R329.24;R345.4;R977.3
DOI:
10.3969/j.issn.1001-1978.2017.07.006
文献标志码:
A
摘要:
目的 研究骨形态蛋白9(BMP9)诱导干细胞成骨分化过程中对PI3K/Akt信号的激活与环氧酶-2(COX-2)的关系。方法 利用组织化学染色法、化学发光法或定量PCR检测碱性磷酸酶(ALP)的水平,Western blot检测骨桥素(OPN)、骨钙素(OCN)、COX-2、Akt1/2和磷酸化Akt1/2(p-Akt1/2)水平,RT-PCR检测COX-2的表达,用免疫组化或免疫荧光分别检测COX-2的表达水平以及Akt1/2的磷酸化水平,用茜素红染色检测钙盐沉积。结果 BMP9在C2C12细胞中明显增加ALP活性,促进OPN和OCN表达以及钙盐沉积; BMP9对Akt1/2总蛋白无明显影响,能明显增加Akt1/2磷酸化水平, PI3K抑制剂呈浓度依赖性减弱BMP9诱导ALP活性增加。BMP9在C2C12细胞中促进COX-2表达,抑制COX-2明显减弱BMP9在C2C12细胞中诱导ALP活性增加和钙盐沉积的作用。外源性过表达COX-2促进BMP9诱导p-Akt1/2水平增加,而抑制COX-2则明显减弱BMP9诱导Akt1/2磷酸化水平增加。结论 BMP9在诱导干细胞成骨化过程中对PI3K/Akt信号的激活可能与其促进COX-2表达有关。
Abstract:
Aim To investigated the possible effect of COX-2 on the BMP9-induced activation of PI3K/Akt signal in progenitor cells.Methods The activity of alkaline phosphatase(ALP)was measured using histochemical staining or chemiluminescence. The mRNA level of ALP was determined using real-time PCR assay. The protein levels of osteopontin(OPN), osteocalcin(OCN), COX-2, Akt1/2 and phosphorylated Akt1/2 were detected by Western blot. The mRNA level of COX-2 was assayed with RT-PCR, and the mineralization was measured with Alizarin Red staining.Results The ALP activity was apparently increased by BMP9 in C2C12 cells, as well as the protein level of OPN and OCN. The mineralization was also markedly induced by BMP9 in C2C12 cells. BMP9 increased the level of phosphorylated Akt1/2 greatly, although no substantial effect was observed on total protein level of Akt1/2. The BMP9-induced ALP activity was dramatically decreased by the inhibitor of PI3K. The mRNA and protein level of COX-2 were both increased by BMP9 in C2C12cells, and the BMP9-induced ALP activity and mineralization were greatly attenuated by the inhibitor of COX-2. The BMP9-induced phosphorylation level of Akt1/2 was increased by the exogenous expression of COX-2, but decreased by the inhibitor of COX-2.Conclusion Activation of PI3K/Akt signaling may be a critical event in BMP9-induced osteogenic differentiation, and this process may be mediated by the BMP9-upregulated COX-2 in stem cells at least.

参考文献/References:

[1] Luu H H, Song W X, Luo X, et al. Distinct roles of bone morphogenetic proteins in osteogenic differentiation of mesenchymal stem cells[J].J Orthop Res, 2007, 25(5):665-77.
[2] Liu Y, Liu Y, Zhang R, et al. All-trans retinoic acid modulates bone morphogenic protein 9-induced osteogenesis and adipogenesis of preadipocytes through BMP/Smad and Wnt/β-catenin signaling pathways[J]. Int J Biochem Cell Biol, 2014, 47:47-56.
[3] Lamplot J D, Qin J, Nan G, et al. BMP9 signaling in stem cell differentiation and osteogenesis[J]. Am J Stem Cells, 2013, 2(1):1-21.
[4] Li L, Dong Q, Wang Y, et al. Hedgehog signaling is involved in the BMP9-induced osteogenic differentiation of mesenchymal stem cells[J]. Int J Mol Med, 2015, 35(6):1641-50.
[5] Zhang W, Deng Z L, Chen L, et al. Retinoic acids potentiate BMP9-induced osteogenic differentiation of mesenchymal progenitor cells[J].PLoS One, 2010, 5(7):e11917.
[6] Yang S X, Polley E, Lipkowitz S. New insights on PI3K/AKT pathway alterations and clinical outcomes in breast cancer[J]. Cancer Treat Rev, 2016, 45:87-96.
[7] Boban A, Lambert C, Hermans C. Is the cardiovascular toxicity of NSAIDS and COX-2 selective inhibitors underestimated in patients with haemophilia?[J].Crit Rev Oncol Hematol, 2016, 100:25-31.
[8] Wang J H, Liu Y Z, Yin L J, et al. BMP9 and COX-2 form an important regulatory loop in BMP9-induced osteogenic differentiation of mesenchymal stem cells[J]. Bone, 2013, 57(1): 311-21.
[9] 黄 军,刘映孜,袁霜雪,等. 环氧酶-2在骨形态蛋白9诱导间充质干细胞骨向分化中的作用研究[J]. 中国药理学通报, 2014, 30(7):1006-11.
[9] Huang J, Liu Y Z, Yuan S X, et al. Study on the role of COX-2 in BMP9 induced osteogenic differentiation in mesenchymal stem cells[J]. Chin Pharmacol Bull, 2014, 30(7):1006-11.
[10] Yun K, Im S H. Lef1 regulates COX-2 transcription in chondrocytes[J]. Biochem Biophys Res Commun, 2007, 364(2):270-5.
[11] Luo J, Deng Z L, Luo X,et al. A protocol for rapid generation of recombinant adenoviruses using the AdEasy system[J].Nat Protoc, 2007, 2(5):1236-47.
[12] Kim M, Choe S. BMPs and their clinical potentials[J]. BMB Rep, 2011, 44(10):619-34.
[13] Akahashi K, Ogura N, Aonuma H, et al. Bone morphogenetic protein 6 stimulates mineralization in human dental follicle cells without dexamethasone[J]. Arch Oral Biol, 2013, 58(6):690-8.
[14] Huang J, Yuan S X, Wang D X, et al. The role of COX-2 in mediating the effect of PTEN on BMP9 induced osteogenic differentiation in mouse embryonic fibroblasts[J]. Biomaterials, 2014, 35(36):9649-59.
[15] Wang Z, Bhattacharyya T. Trends of non-union and prescriptions for non-steroidal anti-inflammatory drugs in the United States, 1993-2012[J]. Acta Orthop, 2015, 86(5):632-7.
[16] Zhang X, Schwarz E M, Young D A, et al. Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair[J]. J Clin Invest, 2002, 109(11):1405-15.
[17] 周龙洋,杨秋珺,牟钰钦,等.美洛昔康对骨形态发生蛋白9诱导间充质干细胞骨向分化的抑制作用[J].中国药理学与毒理学杂志,2012,26(3):321-6.
[17] Zhou L Y, Yang Q J, Mou Y Q, et al. The inhibitory effect of meloxicam on bone morphogenetic protein 9 induced osteogenic differentiation in mesenchymal stem cells[J]. Chin J Pharm Toxicol, 2012, 26(3):321-6.
[18] Chien P T, Lin C C, Hsiao L D, et al. c-Src/Pyk2/EGFR/PI3K/Akt/CREB-activated pathway contributes to human cardiomyocyte hypertrophy: Role of COX-2 induction[J]. Mol Cell Endocrinol,2015, 409:59-72.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(No 81372120)
作者简介:李 洋(1987-),女,硕士生,研究方向:分子药理学,E-mail:506739201@qq.com;
何百成(1972-),男,博士,教授,研究方向:分子药理学和干细胞生物学,通讯作者,E-mail: hebaicheng99@yahoo.com
更新日期/Last Update: 2017-07-15