Effects of secreted factors in culture medium of annulus fibrosus cells on microvascular endothelial cells: elucidating the possible pathomechanisms of matrix degradation and nerve in-growth in disc degeneration.
Effects of secreted factors in culture medium of annulus fibrosus cells on microvascular endothelial cells: elucidating the possible pathomechanisms of matrix degradation and nerve in-growth in disc degeneration.
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DOI：
10.1016/j.joca.2013.12.008
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发表时间：
2014-02
影响因子：
7
通讯作者：
Vo, N. V.
中科院分区：
文献类型：
--
作者：
Moon, H. J.;Yurube, T.;Lozito, T. P.;Pohl, P.;Hartman, R. A.;Sowa, G. A.;Kang, J. D.;Vo, N. V.
关键词：
测试纤维环细胞(AFCs)和内皮细胞(ECs)之间的相互作用是否会破坏基质的动态平衡并刺激神经介质的产生。人微血管内皮细胞在退变的人手术标本的AF细胞培养的条件培养液中培养。采用定量逆转录聚合酶链式反应(qRT-PCR)、Western和免疫荧光法检测内皮细胞基质金属蛋白酶(MMPs)和血小板衍生生长因子(PDGF)的表达。用ELISA法检测细胞培养上清液中血管内皮生长因子(VEGF)、白介素8(IL-8)和神经生长因子(NGF)的含量。采用定量逆转录聚合酶链式反应(qRT-PCR)检测EC条件培养液中培养的AFCs中I型、II型胶原和聚集素的表达水平，以确定ECs对AFCs的作用。与单纯培养的内皮细胞相比，暴露于AFC条件培养液中的内皮细胞表达侵袭性内皮细胞表型的关键生物标志物--基质金属蛋白酶-2(2x)、基质金属蛋白酶-13(4x)、血小板衍生生长因子-B(1.5-2x)和神经生长因子(24.9±15.2pg/mLvs.0)的mRNA和蛋白水平均较高。用EC培养条件培养液处理AF细胞后，II型胶原的表达减少了两倍。在未处理的房颤细胞条件培养液中也检测到大量的促血管生成因子IL-8(396.7±302.0 pg/mL)和血管内皮生长因子(756.2±375.9 pg/mL)。退变的椎间盘中的AFCs分泌因子，刺激EC产生已知的诱导基质降解、血管生成和神经支配的因子。IL-8和VEGF可能是AFC分泌的因子，介导促血管生成刺激，常参与椎间盘退变的发生发展。
To test whether the interaction between annulus fibrosus cells (AFCs)and endothelial cells (ECs) disrupts matrix homeostasis and stimulates production of innervation mediators. Human microvascular ECs were cultured in the conditioned media of AF cell culture derived from degenerated human surgical specimen. Matrix-metalloproteinases (MMPs) and platelet-derived growth factor (PDGF) of ECs of this culture were analyzed by qRT-PCR, Western, and immunofluorescence. Vascular endothelial growth factor (VEGF), Interleukin-8 (IL-8), and nerve growth factor (NGF) in the media of this cell culure were assayed by ELISA. To determine the effects of ECs on AFCs, qRT-PCR was performed to determine mRNA levels of collagen I, II and aggrecan in AFCs cultured in EC conditioned media. Compared to ECs cultured in naïve media, ECs exposed to AFC conditioned media expressed higher mRNA and protein levels of key biomarkers of invasive endothelial cell phenotype, MMP-2 (2x), MMP-13 (4x), and PDGF-B (1.5-2x), and NGF (24.9 ±15.2 pg/mL vs. 0 in naïve media). Treatment of AF cells with EC culture conditioned media decreased collagen type II expression two fold. Considerable quantities of pro-angiogenic factors IL-8 (396.7 ± 302.0 pg/mL) and VEGF (756.2 ± 375.9 pg/mL) were also detected the conditioned media of untreated AF cell culture. AFCs from degenerated discs secreted factors which stimulated EC production of factors known to induce matrix degradation, angiogenisis, and innervation. IL-8 and VEGF maybe the secreted factors from AFCs which mediate a pro-angiogenic stimulus often implicated in the development of disc degeneration.
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影响因子：
2.4
作者：
Michalek, A. J.;Gardner-Morse, M. G.;Iatridis, J. C.
通讯作者：
Iatridis, J. C.
影响因子：
2.8
作者：
Lee, Jae Man;Song, Ji Ye;Shin, Dong Eun
通讯作者：
Shin, Dong Eun
影响因子：
168.9
作者：
Freemont, AJ;Peacock, TE;Jayson, MIV
通讯作者：
Jayson, MIV
影响因子：
7.3
作者：
Le Maitre, CL;Freemont, AJ;Hoyland, JA
通讯作者：
Hoyland, JA
影响因子：
3
作者：
Nasto LA;Seo HY;Robinson AR;Tilstra JS;Clauson CL;Sowa GA;Ngo K;Dong Q;Pola E;Lee JY;Niedernhofer LJ;Kang JD;Robbins PD;Vo NV
通讯作者：
Vo NV