Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts

doi:10.3969/j.issn.2095-4344.2016.06.003

Chinese Journal of Tissue Engineering Research ›› 2016, Vol. 20 ›› Issue (6): 777-784.doi: 10.3969/j.issn.2095-4344.2016.06.003

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Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts

Yang Zhi-lie^{1, 2}, Wang Cheng-long², Zhao Dong-feng², Chang Jun-li², Yang Cheng¹, Yang Yan-ping², Wang Yong-jun²

¹Institute of Integrated Chinese and Western Medicine, Binzhou Medical University, Yantai 264003, Shandong Province, China; ²Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China

Received:2016-01-04 Online:2016-02-05 Published:2016-02-05
Contact: Yang Yan-ping, Researcher, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China Corresponding author: Wang Yong-jun, Professor, Doctoral supervisor, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
About author:Yang Zhi-lie, Master, Institute of Integrated Chinese and Western Medicine, Binzhou Medical University, Yantai 264003, Shandong Province, China; Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Supported by:
the National Natural Science Foundation of China, No. 81373667, 81202708; the National Basic Research Program of China (973 Program), No. 2010CB530400; a grant for the “085” First-Class Disciplines Construction of Shanghai University of Traditional Chinese Medicine, No. 085ZY1217,

085ZY1204; the Innovation Team Development Program of Ministry of Education in China,

No. IRT1270; the Dragon Medical Scholar and Team Program of the National TCM Clinical Research Base, No. LYTD-23; the Sail Program of Shanghai Science and Technology Department, No. 15YF1412200; the Natural Science Foundation of Longhua Hospital of Shanghai University of Traditional Chinese Medicine, No. 14YR03

Abstract

Abstract:

BACKGROUND: Osteoporosis caused by chemotherapy has become one of the serious side effects that impact the skeletal system. Icariin shows a strong anti-osteoporosis activity, which can have protective effect on osteoporosis induced by chemotherapy.
OBJECTIVE: To study the protective effect and mechanism of icariin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts.
METHODS: MTT assay and alkaline phosphatase (ALP) staining were used to determine the optimal protective concentration of icariin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts. mRNA expressions of osteoblast-specific transcription factors, OC, ALP, Runx2, and Wnt/β-catenin signaling pathway target genes, β-catenin, C-Myc, cyclin D1, were determined using RT-PCR method at different time after intervention with the optimal concentration of icariin. Expressions of Runx2, β-catenin, c-Myc, cyclin D1 regulated by the optimal concentration of icariin were detected using western blot assay at the protein level.
RESULTS AND CONCLUSION: Cell viability and ALP activity decreased significantly in the cyclophosphamide group compared with the control group, but there was no significant difference in cell viability between icariin group and cyclophosphamide group. Icariin at 100 μmol/L showed the best protective effect against cyclophosphamide-induced obstacle of osteogenic differentiation of bone marrow mesenhymal stem cells. Compared with the control group, cyclophosphamide chemotherapy reduced the expressions of ALP, OC, Runx2 at mRNA level and Runx2 at protein level, weakened the expressions of β-catenin, cyclin D1 at mRNA level and active β-catenin, Cyclin D1, c-myc at protein level, and increased the expression of DKK1. Compared with the cyclophosphamide group, 100 μmol/L icariin increased the expression of osteoblast-specific transcription factors and Wnt/β-catenin signaling pathway genes at mRNA and protein levels, and reduced the expression of DKK1 protein. These results show that cyclophosphamide can lead to osteogenic differentiation disorder of mouse bone marrow mesenchymal stem cells, and in contrast, icariin shows a protective effect and its optimal intervention concentration is 100 μmol/L. Additionally, the protective role of icariin is probably related to activation of Wnt/β-catenin signal pathway.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Yang Zhi-lie, Wang Cheng-long, Zhao Dong-feng, Chang Jun-li, Yang Cheng, Yang Yan-ping, Wang Yong-jun . Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(6): 777-784.

Figures/Tables 4

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Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts

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