Inhibitory effect of all ̄trans retinoic acid on osteogenic differentiation of mouse embryonic palate mesenchymal cells and its possible mechanism
10.3867/j.issn.1000 ̄3002.2015.04.061
- VernacularTitle:全反式维 A 酸对小鼠胚胎腭间充质细胞成骨分化的抑制作用及其机制
- Author:
Mu CHEN
;
Xu YANG
;
Zhengming LI
;
Xue LIU
;
Weicai WANG
;
Hongzhang HUANG
- Publication Type:Journal Article
- Keywords:
all ̄trans retinoic acid;
embryonic palatal mesenchyme cells;
bone morphogenetic protein receptor
- From:
Chinese Journal of Pharmacology and Toxicology
2015;(5):836-841
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the effect and related mechanism of all ̄trans retinoic acid (atRA) exposure on osteogenic differentiation of mouse embryonic palate masenchymal cells MEPM. METHODS MEPM were cultured in osteogenic medium (OM) with atRA 0.1 and 1.0 μmol??L-1 for 1, 3,5, 7 and 9 d. MTT assay was performed to measure the cell viability. The alkaline phosphatase (ALP) activity was measured by chemical colorimetry. The cells were stained using the Von ̄Kossa technique to detect the formation of mineralization nodules after 21 d of culture. RT ̄PCR was performed to determine expression Runx2, osteopontin, bone morphogenetic protein receptor ( Bmpr) 1b, Bmpr2 and Smad5 mRNA. RESULTS The result of MTT on 9 d showed that, compared with normal control group, the cell viability of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups decreased significantly(P<0.01). Compared with normal control group, ALP activity of OM group increased significantly(P<0.05), while the ALP activity of OM+atRA 0.1 and 1.0 μmol??L-1 groups was lower than OM group(P<0.05). On 21 d, the Von ̄Kossa stai ̄ning results showed that the percentage of mineralization nodules formation of OM+atRA 1.0 μmol??L-1 group was (3.65±1.24)%, which was significantly lower than that of OM group(10.33±2.29)%(P<0. 05). On 9 d, the relative Run expression of OM group was the highest one in the four groups, while at ̄RA 1.0 μmol??L-1 treatment negatively regulated 20% in comparsion with OM group(P<0.05). Compared with normal control group, the mRNA expression of osteopontin of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups increased significantly(P<0.05); BDNF mRNA expression of OM group was 2.6 ̄fold to normal control group, while that of OM+atRA 1.0 μmol??L-1 group was 33% to OM group(P<0.05) . The level of Smad5 mRNA of OM+atRA 1.0 μmol??L-1 group was significantly lower than that of OM group(P<0.05). CONCLUSION atRA Might inhibit osteogenic differentiation of MEPM by down ̄regulated the expression of Bmpr1b.