title: Naringin promotes osteogenic differentiation of bone marrow stromal cells by up-regulating Foxc2 expression via the IHH signaling pathway.
abstract: 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.
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
authors.forename: Fei-Xiang
authors.initials: FX
authors.lastname: Lin
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
authors.forename: Shi-Xin
authors.initials: SX
authors.lastname: Du
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
authors.forename: De-Zhong
authors.initials: DZ
authors.lastname: Liu
authors.affiliation: 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.
authors.forename: Qin-Xiao
authors.initials: QX
authors.lastname: Hu
authors.affiliation: 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.
authors.forename: Guo-Yong
authors.initials: GY
authors.lastname: Yu
authors.affiliation: 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.
authors.forename: Chu-Cheng
authors.initials: CC
authors.lastname: Wu
authors.affiliation: 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.
authors.forename: Gui-Zhou
authors.initials: GZ
authors.lastname: Zheng
authors.affiliation: 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.
authors.forename: Da
authors.initials: D
authors.lastname: Xie
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
authors.forename: Xue-Dong
authors.initials: XD
authors.lastname: Li
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Shantou University Medical College Shantou 515041, Guangdong, P. R. China.
authors.forename: Bo
authors.initials: B
authors.lastname: Chang
keywords: Naringin; bone marrow stromal cells (BMSCs); forkhead box C2 (Foxc2); indian hedgehog (IHH); osteogenic differentiation; osteoporosis