Effect of Irisin on osteogenic differentiation and Sost of rat bone marrow mesenchymal stem cells

doi:10.3877/cma.j.issn.1674-1366.2019.03.003

Abstract

Abstract:

Objective

To investigate the effect of Irisin on osteogenic differentiation and Sost of rat bone marrow mesenchymal stem cells (BMSCs) .

Methods

Irisin was set to have different concentrations (0, 80, 100, 120 ng/mL) , and the optimal concentration of Irisin was determined by CCK-8 method. The BMSCs cultured with Irisin was the experimental group, while the control group was cultured without Irisin. The alizarin red and von Kossa were stained on the 3rd, 7th and 14th day to observe the positive region of osteogenic differentiation and the expression intensity. The mRNA and protein expression levels of the bone-related genes were detected by RT-PCR and Western bolt at 3rd and 10th days. The One-Way ANOVA and t-test were applied to perform the statistical analysis of the data.

Results

(1) Irisin at a concentration of 100 ng/mL promoted the proliferation of rat BMSCs significantly after cultured three days (OD=0.951; F=102.52, P<0.05) . (2) The depth of staining and the size as well as the number of calcium nodules in the experimental group were significantly higher than those in the control group. (3) Compared with the control group, the mRNA and protein expression levels of the bone-related genes: ALP, Lrp5, BMP2 and Smad1 were significantly increased, but the expression levels of Sost mRNA and protein were significantly decreased (P<0.001) .

Conclusions

(1) Irisin can inhibit the expression of the Sost gene (2) Irisin can promote the proliferation and osteogenic differentiation of rat BMSCs in vitro. (3) Wnt and BMP signaling pathway may play a role in Irisin promoting osteogenic differentiation of rat BMSCs.

Key words: Bone marrow, mesenchymal stem cells, Signal transduction, Irisin, Sclerostin

Mengting Tian, Jing Liu, Xuemin Zeng, Wenjing Zhang, Xiangzhen Han, Huiyu He. Effect of Irisin on osteogenic differentiation and Sost of rat bone marrow mesenchymal stem cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2019, 13(03): 144-150.

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Figures/Tables 9

References 24

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基因名称	引物序列
GAPDH	F:5′-GGCACAGTCAAGGCTGAGAATG-3′
GAPDH	R:5′-ATGGTGGTGAAGACGCCAGTA-3′
ALP	F:5′-AACGTGGCCAAGAACATCATCA-3′
ALP	R:5′-TGTCCATCTCCAGCCGTGTC-3′
BMP2	F:5′-CAACACCGTGCTCAGCTTCC-3′
BMP2	R:5′-TTCCCACTCATTTCTGAAAGTTCC-3′
Smad1	F:5′-GGTACCAATGACATTTATTTTCC-3′
Smad1	R:5′-CCTCGAGTTTATACATGAAATAGG-3′
Lrp5	F:5′-CTTCATCCACCGTGCCAACCTG-3′
Lrp5	R:5′-TCTGCCAGTCTGTCCAGTAGAGTG-3′
Sost	F:5′-GCCTTGCCTGCCTGCCTGTAC-3′
Sost	R:5′-CCGGTTCATGGTCTGGTTGTTCTC-3′

基因名称	引物序列
GAPDH	F:5′-GGCACAGTCAAGGCTGAGAATG-3′
GAPDH	R:5′-ATGGTGGTGAAGACGCCAGTA-3′
ALP	F:5′-AACGTGGCCAAGAACATCATCA-3′
ALP	R:5′-TGTCCATCTCCAGCCGTGTC-3′
BMP2	F:5′-CAACACCGTGCTCAGCTTCC-3′
BMP2	R:5′-TTCCCACTCATTTCTGAAAGTTCC-3′
Smad1	F:5′-GGTACCAATGACATTTATTTTCC-3′
Smad1	R:5′-CCTCGAGTTTATACATGAAATAGG-3′
Lrp5	F:5′-CTTCATCCACCGTGCCAACCTG-3′
Lrp5	R:5′-TCTGCCAGTCTGTCCAGTAGAGTG-3′
Sost	F:5′-GCCTTGCCTGCCTGCCTGTAC-3′
Sost	R:5′-CCGGTTCATGGTCTGGTTGTTCTC-3′

组别	ALP	Lrp5	Sost	BMP2	Smad1
对照组	1.00 ± 0.11	0.98 ± 0.12	1.02 ± 0.19	1.00 ± 0.09	1.86 ± 0.26
实验组	1.75 ± 0.51	1.78 ± 0.23	0.64 ± 0.13	1.41 ± 0.26	1.04 ± 0.34
t值	4.276	9.057	4.886	4.571	6.176
P值	0.001	0.001	0.001	0.001	0.001

组别	ALP	Lrp5	Sost	BMP2	Smad1
对照组	1.00 ± 0.11	0.98 ± 0.12	1.02 ± 0.19	1.00 ± 0.09	1.86 ± 0.26
实验组	1.75 ± 0.51	1.78 ± 0.23	0.64 ± 0.13	1.41 ± 0.26	1.04 ± 0.34
t值	4.276	9.057	4.886	4.571	6.176
P值	0.001	0.001	0.001	0.001	0.001

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