Lin Fei-Xiang, Du Shi-Xin, Liu De-Zhong, Hu Qin-Xiao, Yu Guo-Yong, Wu Chu-Cheng, Zheng Gui-Zhou, Xie Da, Li Xue-Dong, Chang Bo
Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical CollegeShantou 515041, Guangdong, P. R. China; Department of Orthopedics, The Affiliated Luohu Hospital of Shenzhen UniversityShenzhen 518000, Guangdong, P. R. China.
Am J Transl Res. 2016 Nov 15;8(11):5098-5107. eCollection 2016.
Naringin is an active compound extracted from Rhizoma Drynariae, and studies have revealed that naringin can promote proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs). In this study, we explored whether naringin could promote osteogenic differentiation of BMSCs by upregulating Foxc2 expression via the Indian hedgehog (IHH) signaling pathway. BMSCs were cultured in basal medium, basal medium with naringin, osteogenic induction medium, osteogenic induction medium with naringin and osteogenic induction medium with naringin in the presence of the IHH inhibitor cyclopamine (CPE). We examined cell proliferation by using a WST-8 assay, and differentiation by Alizarin Red S staining (for mineralization) and alkaline phosphatase (ALP) activity. In addition, we detected core-binding factor α1 (Cbfα1), osteocalcin (OCN), bone sialoprotein (BSP), peroxisome proliferation-activated receptor gamma 2 (PPARγ2) and Foxc2 expression by using RT-PCR. We also determined Foxc2 and IHH protein levels by western blotting. Naringin increased the mineralization of BMSCs, as shown by Alizarin red S assays, and induced ALP activity. In addition, naringin significantly increased the mRNA levels of Foxc2, Cbfα1, OCN, and BSP, while decreasing PPARγ2 mRNA levels. Furthermore, the IHH inhibitor CPE inhibited the osteogenesis-potentiating effects of naringin. Naringin increased Foxc2 and stimulated the activation of IHH, as evidenced by increased expression of proteins that were inhibited by CPE. Our findings indicate that naringin promotes osteogenic differentiation of BMSCs by up-regulating Foxc2 expression via the IHH signaling pathway.
柚皮苷是从骨碎补中提取的一种活性化合物,研究表明柚皮苷可促进骨髓间充质干细胞(BMSCs)的增殖和成骨分化。在本研究中,我们探讨了柚皮苷是否能通过印度刺猬因子(IHH)信号通路上调Foxc2表达来促进BMSCs的成骨分化。将BMSCs培养于基础培养基、含柚皮苷的基础培养基、成骨诱导培养基、含柚皮苷的成骨诱导培养基以及在存在IHH抑制剂环杷明(CPE)的情况下含柚皮苷的成骨诱导培养基中。我们使用WST - 8法检测细胞增殖,并通过茜素红S染色(用于矿化)和碱性磷酸酶(ALP)活性检测分化情况。此外,我们通过RT - PCR检测核心结合因子α1(Cbfα1)、骨钙素(OCN)、骨唾液蛋白(BSP)、过氧化物酶体增殖物激活受体γ2(PPARγ2)和Foxc2的表达。我们还通过蛋白质印迹法测定Foxc2和IHH蛋白水平。茜素红S检测显示,柚皮苷增加了BMSCs的矿化,并诱导了ALP活性。此外,柚皮苷显著增加了Foxc2、Cbfα1、OCN和BSP的mRNA水平,同时降低了PPARγ2 mRNA水平。此外,IHH抑制剂CPE抑制了柚皮苷的成骨增强作用。柚皮苷增加了Foxc2并刺激了IHH的激活,这可通过CPE抑制的蛋白表达增加来证明。我们的研究结果表明,柚皮苷通过IHH信号通路上调Foxc2表达来促进BMSCs的成骨分化。
Shanghai Kou Qiang Yi Xue. 2014-6
Molecules. 2023-4-25
Front Bioeng Biotechnol. 2022-2-18
Med Sci Monit. 2019-4-28
Arthritis Res Ther. 2014-7-17
Mol Biol Cell. 2012-7-25
Arthritis Res Ther. 2012-7-20
Proc Natl Acad Sci U S A. 2011-8-22