Effects of Orexin-1 Receptor Inhibitor on Proliferation and Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells / 医药导报

ABSTRACT

Objective To observe the effects of orexin-1 receptor inhibitor on proliferation and estrogenic differentiation of bone marrow mesenchymal stem cells ( BMSCs) . Methods BMSCs were isolated and cultured from Sprague-Dawley rats, and orexin-1 receptor inhibitor was prepared to different concentration of solutions10-2 , 10-3 , 10-4 , 10-5 and 10-6 mol·L-1 . Then, the cells at passage 3 were cultured in orexin-1 receptor inhibitor solutions. The cell growth, alkaline phosphatase activity and osteocalcin secretion were measured by MTT, PNPP and ELISA, respectively. The mRNA expression levels of Runx2 and COL1A1 were detected by real-time quantitative PCR. Results The best concentration of orexin-1 receptor inhibitor for the proliferation of BMSCs was 1 × 10-4 mol·L-1 . Orexin-1 receptor inhibitor promoted the proliferation at concentration of 10-6 , 10-5 , 10-4 mol·L-1 in 3, 5 and 7 days. Orexin-1 receptor inhibitor at concentration of 10-6 , 10-5 and 10-4 mol·L-1 for 5 days significantly stimulated ALP activity. Orexin-1 receptor inhibitor significantly up-regulated Runx2 and COL1A1 mRNA expression at concentration of 10-6, 10-5, 10-4 mol·L-1 for 5 days (P<0.05). Conclusion Orexin-1 receptor inhibitor can promote BMSCs proliferation and stimulated ALP activity and osteocalcin secretion.