Effects of puerarin on proliferation of osteoblasts and Runx2-targeting miRNAs
10.3969/j.issn.1001-1978.2016.10.024
- VernacularTitle:葛根素对成骨细胞增殖能力及靶向Runx2的miRNA的影响
- Author:
Yingying ZHANG
;
Jianbin ZHOU
;
Xiangwei ZENG
;
Fengming ZHAO
;
Guangdong LIU
;
Xiuqin ZHAN
- Publication Type:Journal Article
- Keywords:
puerarin;
osteoblast;
proliferation;
alka-line phosphatase;
Runx2;
miR-204;
dual luciferase reporter gene
- From:
Chinese Pharmacological Bulletin
2016;32(10):1457-1462
- CountryChina
- Language:Chinese
-
Abstract:
Aim To study proliferation capacity of cell and the target relationship between microRNA and Runx2 after effect of puerarin on osteoblasts MC3 T3-E1 . Methods The proliferation capacity of cell was detected by MTT after effect of puerarin on osteoblasts MC3 T3-E1 . The vitality of osteoblasts was detected by activity of alkaline phosphatase. The expression level of mRNA and protein of Runx2 were detected by real-time quantitative PCR and Western blot. The result of miRNA expression spectrum was compared with the predicted result to determine the Runx2-targeting miR-NAs. The expression levels of miRNAs possiby targeted to Runx2 were detected by real-time quantitative PCR. The RhoE 3′UTR vector and RhoE mut 3′UTR vector were constructed. miRNA-204 mimics and miRNA-204 NC were synthetised. The target genes were verified by dual luciferase report gene assay. Results After osteo-blasts treated with puerarin, proliferation capacity and activity of cells were enhanced , expression levels of mRNA and protein of Runx2 were both increased , the expression levels of miRNA-204 and miRNA-344 f-5 p were declined, the expression levels of miRNA-2861 was increased,the expression levels of miRNA-23a-5p, miRNA-770-5 p and miRNA-871-5 p showed no obvious change. According to the results of dual luciferase re-porter gene method after cell transfection of 48 h, only set of 3′UTR Runx2+mimics the miRNA-204 of fluo-rescein protein expression level decreased significantly, showing only the miRNA-204 inhibits Runx2 3′UTR report gene expression. Conclusion Puerarin pro-motes the proliferation of osteoblasts and regulates the miRNAs which possibly target to Runx2 .
