Aqueous Fraction of Huogu Formula Promotes Osteogenic Differentiation of Bone Marrow Stromal Cells Through the BMP and Wnt Signaling Pathways.
Our recent studies have shown that Huogu (HG) formula was effective both in clinic experience and in experimental osteonecrosis of the femoral head (ONFH). Given that defective of bone marrow stromal cells (MSCs) contribute to the development of osteonecrosis and MSCs show enormous potential in the treatment of ONFH, especially to aging people. How HG impacts the differentiation of MSCs and what is the underlying cellular and molecular mechanism remains largely unknown. Here, we found that an aqueous fraction of HG (HGA) significantly increased the alkaline phosphatase (ALP) activity, mineralized nodules, and migration of MSCs in a dose-dependent manner. Meanwhile, HGA could enhance the mRNA and protein expression of Runt-related transcription factor 2 (Runx2), Alp, Bmp2, osteocalcin (Ocn), and Osterix (Osx). Further investigation of the molecular mechanisms revealed that HGA treatment obviously increased expression, secretion, and activation of bone morphogenetic protein (BMP) 2 and β-catenin, two key regulators of the BMP or Wnt signaling pathway. Furthermore, osteogenic differentiation of MSCs could be blocked by using pharmacological inhibitors for these signaling pathways such as Noggin and Dkk-1. Besides, HGA could inhibit adipogenic differentiation of MSCs. Our study reveals that HGA promotes the osteogenesis of MSCs via the BMP and Wnt signaling pathways. Our findings provide mechanistic insights into the role of HG in treating ONFH.
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Xiangying
X
Kong
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Xiaomin
X
Li
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Cun
C
Zhang
2 Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University , Beijing, China .|3 Beijing Key Laboratory of Emerging Infectious Diseases , Beijing, China .
Liuluan
L
Zhu
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Hongye
H
Wan
4 Wangjing Hospital , China Academy of Chinese Medical Sciences, Beijing, China .
Jia
J
Zhu
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Cuiling
C
Liu
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Hongchang
H
Su
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Qingxia
Q
Qin
4 Wangjing Hospital , China Academy of Chinese Medical Sciences, Beijing, China .
Weiheng
W
Chen
1 Institute of Chinese Materia Medica , China Academy of Chinese Medical Sciences, Beijing, China .
Na
N
Lin
D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000502:physiology*; D002352:Carrier Proteins / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D000099176:Noggin Protein
D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D050659:Core Binding Factor Alpha 1 Subunit; D004365:Drugs, Chinese Herbal; D036341:Intercellular Signaling Peptides and Proteins; D000099176:Noggin Protein; C531384:Dkk1 protein, rat; C493717:Runx2 protein, rat
BMP; Wnt; aqueous fraction of HG