Smooth muscle cells of human veins show an increased response to injury at valve sites.
Smooth muscle cells of human veins show an increased response to injury at valve sites.
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DOI：
10.1016/j.jvs.2017.03.447
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发表时间：
2018-05
影响因子：
4.3
通讯作者：
Kenagy RD
中科院分区：
文献类型：
--
作者：
Kikuchi S;Chen L;Xiong K;Saito Y;Azuma N;Tang G;Sobel M;Wight TN;Kenagy RD
静脉瓣膜至关重要，但容易受伤、血栓形成和纤维化。我们比较了瓣膜窦和非瓣膜部位平滑肌细胞 (SMC) 的行为和基因表达，以阐明与静脉瓣膜相关的生物学差异。制备新鲜人隐静脉的组织外植体，并测量瓣膜窦与非瓣膜窦区域外植体中 SMC 的迁移情况。通过 Ki67 和 TUNEL 染色测定 SMC 的增殖和死亡。通过细胞计数并使用微趋化室来确定传代的瓣膜 SMC 与非瓣膜 SMC 的增殖和迁移。通过 RNA 测序确定瓣膜与非瓣膜内膜/中膜的整体基因表达。与非瓣膜 SMC 相比，瓣膜 SMC 在组织外植体中表现出更大的增殖（4 天时 Ki67 阳性核分别为 19.3±5.4% 与 6.8±2.0%；平均值 ± SEM，5 个静脉；P<.05）。对于迁移也是如此（第 6 天时，迁移 SMC/外植体分别为 18.2±2.7 和 7.5±3.0；24 个静脉，15 个外植体/静脉；P<.0001）。细胞死亡没有差异（4 天、5 条静脉时，TUNEL 阳性细胞分别为 39.6±16.1% 和 41.5±16.0%）。培养的瓣膜 SMC 对 PDGF-BB 的反应也比非瓣膜 SMC 增殖更快（分别是对照的 2.9±0.2 倍与 2.1±0.2 倍；P<.001；N=5 对细胞）。对于迁移也是如此（分别为对照的 6.5±1.2 倍与 4.4±0.8 倍；P<.001；N=7 对细胞）。阻断 FGF2 可抑制瓣膜 SMC 的反应增强，但对非瓣膜 SMC 没有影响。外源性 FGF2 增加了瓣膜的迁移，但不增加非瓣膜 SMC 的迁移。与分离的培养细胞不同，组织外植体中 FGF2 的阻断不会阻止瓣膜或非瓣膜 SMC 从外植体的迁移。与非瓣膜内膜/内侧组织（11 条静脉）相比，瓣膜有 37 个基因差异表达。 SEMA3A（差异表达基因之一）的肽介导抑制增加了瓣膜迁移 SMC 的数量，但不增加非瓣膜外植体的迁移 SMC 数量。与非瓣膜相比，瓣膜中的 SMC 具有更高的迁移和增殖率，这可能部分解释了瓣膜中病理病变形成的倾向。虽然 FGF2 在培养的 SMC 中介导这些作用，但在瓣膜壁组织中这些刺激作用的介体仍不清楚，但可能是本研究中发现的差异表达基因之一。这些基因之一，SEMA3A，介导对瓣膜 SMC 损伤反应的瓣膜特异性抑制作用。
Venous valves are essential, but are prone to injury, thrombosis, and fibrosis. We compared the behavior and gene expression of smooth muscle cells (SMCs) in the valve sinus vs non-valve sites to elucidate biological differences associated with vein valves. Tissue explants of fresh human saphenous veins were prepared, and the migration of SMCs from explants of valve sinus vs non-valve sinus areas was measured. Proliferation and death of SMCs was determined by staining for Ki67 and TUNEL. Proliferation and migration of passaged valve vs non-valve SMCs was determined by cell counts and using microchemotaxis chambers. Global gene expression in valve vs non-valve intima/media was determined by RNA-Seq. Valve SMCs demonstrated greater proliferation in tissue explants compared to non-valve SMCs (19.3±5.4% vs. 6.8±2.0% Ki67 positive nuclei at 4 days, respectively; mean ± SEM, 5 veins; P<.05). This was also true for migration (18.2±2.7 vs. 7.5±3.0 migrated SMCs/explant at 6 days, respectively; 24 veins, 15 explants/vein; P<.0001). Cell death was not different (39.6±16.1% vs. 41.5±16.0% TUNEL positive cells, respectively, at 4 days, 5 veins). Cultured valve SMCs also proliferated faster than non-valve SMCs in response to PDGF-BB (2.9±0.2 vs. 2.1±0.2 fold of control, respectively; P<.001; N=5 pairs of cells). This was also true for migration (6.5±1.2 vs. 4.4±0.8 fold of control, respectively; P<.001; N=7 pairs of cells). Blockade of FGF2 inhibited the increased responses of valve SMCs, but had no effect on non-valve SMCs. Exogenous FGF2 increased migration of valve, but not non-valve SMCs. Unlike the isolated, cultured cells, blockade of FGF2 in the tissue explants did not block migration of valve or non-valve SMCs from the explants. 37 genes were differentially expressed by valve compared to non-valve intimal/medial tissue (11 veins). Peptide-mediated inhibition of SEMA3A, one of the differentially expressed genes, increased the number of migrated SMCs of valve, but not non-valve explants. Valve, compared to non-valve, SMCs have greater rates of migration and proliferation, which may in part explain the propensity for pathological lesion formation in valves. While FGF2 mediates these effects in cultured SMCs, the mediators of these stimulatory effects in the valve wall tissue remain unclear, but may be among the differentially expressed genes discovered in this study. One of these genes, SEMA3A, mediates a valve-specific inhibitory effect on the injury response of valve SMCs.
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影响因子：
4.3
作者：
Kenagy RD;Civelek M;Kikuchi S;Chen L;Grieff A;Sobel M;Lusis AJ;Clowes AW
通讯作者：
Clowes AW
影响因子：
15.9
作者：
Bazigou, Eleni;Lyons, Oliver T. A.;Makinen, Taija
通讯作者：
Makinen, Taija
影响因子：
2.8
作者：
Ellis, Stephen G.;Chen, Michael S.;Lytle, Bruce
通讯作者：
Lytle, Bruce
DOI：
10.1161/atvbaha.110.216317
发表时间：
2011-01
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Henke PK;Mitsuya M;Luke CE;Elfline MA;Baldwin JF;Deatrick KB;Diaz JA;Sood V;Upchurch GR;Wakefield TW;Hogaboam C;Kunkel SL
通讯作者：
Kunkel SL
DOI：
10.1152/ajpheart.00495.2015
发表时间：
2015-09-15
影响因子：
4.8
作者：
Batchu, Sri N.;Xia, Jixiang;Korshunov, Vyacheslav A.
通讯作者：
Korshunov, Vyacheslav A.