Suppression of osteogenic differentiation in mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament by a histamine-2-receptor antagonist
Suppression of osteogenic differentiation in mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament by a histamine-2-receptor antagonist
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DOI：
10.1016/j.ejphar.2017.07.013
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发表时间：
2017-09-05
影响因子：
5
通讯作者：
Ishibashi, Yasuyuki
中科院分区：
文献类型：
--
作者：
Liu, Xizhe;Kumagai, Gentaro;Ishibashi, Yasuyuki
间充质干细胞（MSCs）在后纵韧带骨化（OPLL）患者中具有高度的成骨倾向。组胺受体H2（H2R）拮抗剂（H2阻断剂）如法莫替丁可通过一种尚不清楚的机制降低患者的骨化。为了证实MSC表达H2R并阐明H2阻断剂如何抑制成骨分化，我们使用了来自OPLL或脊髓型颈椎病（CSM）患者（对照）的脊柱韧带MSC。用10、30或100 nM法莫替丁处理MSC 7或21天。流式细胞术显示，两组细胞均表达MSC表面标志物CD44、CD90和CD105（> 97.5%），但不表达CD34或CD45（< 2.5%）。免疫印迹显示两组间充质干细胞均表达H2R，但OPLL患者的表达水平更高。实时定量PCR表明，30 nM法莫替丁7天或30或100 nM法莫替丁21天显著抑制H2R表达。然而，组氨酸脱羧酶，在组胺产生的关键酶，并没有改变法莫替丁后，显着。法莫替丁以100 nM处理21天，仅在OPLL衍生的MSC中显著抑制成骨标志物骨钙素（OCN）、骨形态发生蛋白2（BMP 2）和侏儒相关转录因子2（RUNX2）的mRNA表达。免疫印迹显示法莫替丁抑制了OPLL组的BMP 2和OCN以及两组的H2 R和RUNX 2。这些结果表明法莫替丁通过作为H2R拮抗剂抑制OPLL来源的MSC的成骨分化，但也通过降低H2R表达，并支持法莫替丁治疗OPLL的临床用途。
Mesenchymal stem cells (MSCs) in ossification of the posterior longitudinal ligament (OPLL) patients have a high propensity toward osteogenesis. Histamine receptor H2 (H2R) antagonists (H2 blockers) like famotidine decrease ossification in patients, by an unclear mechanism. To confirm that MSCs express H2R and to clarify how H2 blockers suppress osteogenic differentiation, we used spinal-ligament MSCs from patients with OPLL or with cervical spondylotic myelopathy (CSM) (control). The MSCs were treated with 10, 30, or 100 nM famotidine for 7 or 21 days. Flow cytornetry revealed that cells from both groups expressed MSC surface markers CD44, CD90, and CD105 (> 97.5%) but not CD34 or CD45 (< 2.5%). Immunoblotting showed that the MSCs from both groups expressed H2R, but those from OPLL patients expressed it at higher levels. Realtime qPCR indicated the H2R expression was significantly suppressed by 30 nM famotidine for 7 days or by 30 or 100 nM for 21 days. However, histidine decarboxylase, a key enzyme in histamine production, did not change significantly after famotidine addition. Famotidine treatment at 100 nM for 21 days significantly suppressed mRNA expression of the osteogenic markers osteocalcin (OCN), bone morphogenetic protein 2 (BMP2), and runt-related transcription factor 2 (RUNX2) only in OPLL-derived MSCs. Immunoblots showed that famotidine suppressed BMP2 and OCN in the OPLL group and H2R and RUNX2 in both groups. These results suggest famotidine inhibits osteogenic differentiation in OPLL-derived MSCs by acting as an H2R antagonist, but also by decreasing H2R expression, and support the clinical use of famotidine to treat OPLL.