Data Availability StatementThe datasets used and/or analyzed through the current research are available through the corresponding author on reasonable request. by miR-19b-3p mimic transfection and inhibited by miR-19b-3p inhibitor transfection. LncRNA H19 was obviously down-regulated in postmenopausal osteoporosis patients. H19 overexpression significantly decreased cell proliferation and differentiation by down-regulating miR-19b-3p. Moreover, the expression of miR-19b-3p was inhibited, while H19 elvated Rabbit Polyclonal to ARC in 17-estradiol (E2) treated BMSCs in a dose-dependent manner. Conclusion These data were the first to reveal the critical role of H19/miR-19b-3p in postmenopausal osteoporosis, and provided a new therapeutic target for OP. test. Differences between larger groups were analyzed by one-way analysis of variance, followed by Dunnetts test. values less than 0.05 were considered significant. Results MiR-19b-3p is up-regulated in postmenopausal osteoporosis patients and BMP-2-induced BMSCs The expression of miR-19b-3p was first evaluated in the serum of postmenopausal osteoporosis patients and heathy Fraxetin controls by qRT-PCR. As shown in Fig.?1a, the expression of miR-19b-3p was obviously elevated in osteoporosis group as compared with healthy control group (P?0.05). To explore the potential role of miR-19b-3p Fraxetin during osteoblast differentiation, the expression of miR-19b-3p was measured in BMSC stimulated with BMP-2, which has been proved to induce osteoblast differentiation . The results indicated miR-19b-3p was significantly increased in BMP-2 induced MSCs as compared with control cells. Open in a separate window Fig. 1 MiR-19b-3p is up-regulated in postmenopausal osteoporosis patients and BMP-2-induced BMSCs. (a) The expression of miR-19b-3p in the serum of postmenopausal osteoporosis patients and heathy controls were measured by qRT-PCR. Each specimen was repeated three times. (b) Control group, normal BMSC cell; BMP-2 group, BMSC cell treated with 100?ng/mL BMP-2. *P?0.05 versus healthy control group MiR-19b-3p promotes proliferation of BMSCs To determine the effect of miR-19b-3p on cell proliferation, miR-19b-3p mimic or inhibitor was transfected into BMP-2 induced BMSCs. The qRT-PCR results showed a significant increase of miR-19b-3p expression in miR-19b-3p mimic transfection group, and an obvious decrease of miR-19b-3p expression in miR-19b-3p inhibitor transfection group as compared with control group (Fig.?2a). BrdU outcomes indicated that cell proliferation level was raised in miR-19b-3p imitate group considerably, while dramatically dropped in miR-19b-3p inhibitor group in comparison with control group (Fig. ?(Fig.22b). Open up in another windowpane Fig. 2 MiR-19b-3p promotes proliferation of BMSCs. Fraxetin Control group, BMSC cells treated with BMP-2; miR-19b-3p imitate group, BMP-2 treated cells transfected with miR-19b-3p imitate; imitate control group, BMP-2 treated cells transfected with imitate control; miR-19b-3p inhibitor group, BMP-2 treated cells transfected with miR-19b-3p inhibitor; inhibitor control group, BMP-2 treated cells transfected with inhibitor control. (a) The manifestation of miR-19b-3p was measure by qRT-PCR. (b) Cell proliferation price was examined by BrdU assay. *P?0.05 versus healthy control group MiR-19b-3p boost differentiation of BMSCs To judge the result of miR-19b-3p on BMSC differentiation, we measured ALP activity as well as the expression degree of RUNX2, COL1A1 in BMP-2 induced BMSCs. As demonstrated in Fig.?3a, ALP activity was elevated in miR-19b-3p mimic group significantly, while decreased in miR-19b-3p inhibitor group in comparison with control group. Furthermore, proteins manifestation of COL1A1 and RUNX2 had been improved in miR-19b-3p imitate group, whereas impeded in miR-19b-3p inhibitor group in comparison Fraxetin to control group (Fig. ?(Fig.3b,3b, c and d). Open up in another windowpane Fig. 3 MiR-19b-3p increase differentiation of BMSCs. (a) ALP activity was recognized in the supernatant of cells. (b) Proteins manifestation of RUNX2 and COL1A1 had been measured by traditional western blot technique. (c and d) Comparative proteins level was normalized to GAPDH. *P?0.05 versus healthy control group H19 up-regulation elevates cell proliferation and differentiation of BMSCs through mediating miR-19b-3p H19 expression was determined in postmenopausal osteoporosis patients and healthy controls. The outcomes demonstrated a significant loss of H19 manifestation in postmenopausal osteoporosis individuals in comparison to healthful settings (Fig.?4a). We evaluated the expression of H19 in BMP-2 stimulated BMSCs then. The outcomes indicated H19 was considerably reduced in BMP-2 induced BMSCs in comparison to control cells (Fig. ?(Fig.4b).4b). To research the relationship between H19 and miR-19b-3p in BMSCs, we transfected pcDNA3.1-H19 alone or with miR-19b-3p imitate into BMP-2 induced BMSCs. As indicated in Fig. ?Fig.4c,4c, H19 expression was improved in H19 group in comparison with control group significantly. Meanwhile, there is absolutely no factor between H19 group and H19?+?miR-19b-3p imitate group. Additionally, miR-19b-3p manifestation was significantly down-regulated in H19 group compared to control group, and increased in H19?+?miR-19b-3p mimic group compared to H19 group (Fig. ?(Fig.4d).4d). Cell proliferation was decreased in.