title: Hydrostatic pressure promotes the proliferation and osteogenic/chondrogenic differentiation of mesenchymal stem cells: The roles of RhoA and Rac1.
abstract: Our previous studies have shown that hydrostatic pressure can serve as an active regulator for bone marrow mesenchymal stem cells (BMSCs). The current work further investigates the roles of cytoskeletal regulatory proteins Ras homolog gene family member A (RhoA) and Ras-related C3 botulinum toxin substrate 1 (Rac1) in hydrostatic pressure-related effects on BMSCs. Flow cytometry assays showed that the hydrostatic pressure promoted cell cycle initiation in a RhoA- and Rac1-dependent manner. Furthermore, fluorescence assays confirmed that RhoA played a positive and Rac1 displayed a negative role in the hydrostatic pressure-induced F-actin stress fiber assembly. Western blots suggested that RhoA and Rac1 play central roles in the pressure-inhibited ERK phosphorylation, and Rac1 but not RhoA was involved in the pressure-promoted JNK phosphorylation. Finally, real-time polymerase chain reaction (PCR) experiments showed that pressure promoted the expression of osteogenic marker genes in BMSCs at an early stage of osteogenic differentiation through the up-regulation of RhoA activity. Additionally, the PCR results showed that pressure enhanced the expression of chondrogenic marker genes in BMSCs during chondrogenic differentiation via the up-regulation of Rac1 activity. Collectively, our results suggested that RhoA and Rac1 are critical to the pressure-induced proliferation and differentiation, the stress fiber assembly, and MAPK activation in BMSCs.
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China.
authors: Yin-Hua
authors: YH
authors: Zhao
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China.
authors: Xin
authors: X
authors: Lv
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China.
authors: Yan-Li
authors: YL
authors: Liu
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China.
authors: Ying
authors: Y
authors: Zhao
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China.
authors: Qiang
authors: Q
authors: Li
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China. Electronic address: cyj1229@fmmu.edu.cn.
authors: Yong-Jin
authors: YJ
authors: Chen
authors: State Key Laboratory of Military Stomatology, Department of General Dentistry and Emergency, School of Stomatology, Fourth Military Medical University, No. 145 West Changle Road, Xi'an 710032, China. Electronic address: zhangmin@fmmu.edu.cn.
authors: Min
authors: M
authors: Zhang
mesh_terms: D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D015536:Down-Regulation; D005434:Flow Cytometry; D005819:Genetic Markers; D006874:Hydrostatic Pressure; D040542:Mechanotransduction, Cellular; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D020830:rac1 GTP-Binding Protein / Q000378:metabolism / Q000502:physiology*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism / Q000502:physiology*
chemical_list: D005819:Genetic Markers; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein