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Suppression of osteogenic differentiation in mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament by a histamine-2-receptor antagonist.
European Journal of Pharmacology 2017 September 6
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 100nM famotidine for 7 or 21 days. Flow cytometry 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. Real-time qPCR indicated the H2R expression was significantly suppressed by 30nM famotidine for 7 days or by 30 or 100nM for 21 days. However, histidine decarboxylase, a key enzyme in histamine production, did not change significantly after famotidine addition. Famotidine treatment at 100nM 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.
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