Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1832-1836.doi: 10.3969/j.issn.1673-8225.2011.10.026

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Effects of fasudil and RhoA silencing on neural stem cell proliferation

Wang Bing-qian1, Wang Dong2, Zhang Jian-jun2, Wang Yong-xin1, Zhu Guo-hua1, Du Guo-jia1, Dang Mu-ren1   

  1. 1Department of Neurosurgery, First Affiliated Hospital, Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
    2Department of Neurosurgery, Tianjin Fourth Central Hospital, Tianjin  300140, China
  • Received:2010-11-03 Revised:2010-11-26 Online:2011-03-05 Published:2011-03-05
  • Contact: Dang Mu-ren, Chief physician, Doctoral supervisor, Department of Neurosurgery, First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China damrjab@xj.cninfo.net
  • About author:Wang Bing-qian★, Studying for master’s degree, Department of Neurosurgery, First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China ydwangbingqian@126.com
  • Supported by:

    the General Program of Science and Technology Foundation of Health Bureau of Tianjin City, No. 2010ky04*

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Abstract:

BACKGROUND: Rho and its relevant molecules play important roles in growth, differentiation, extension and synapse formation of nerve axons. To block and inhibit RhoA/ROCK pathway can promote the proliferation and growth of neural stem cells.
OBJECTIVE: To observe the effects of Rho kinase inhibitor fasudil and RNAi-mediated RhoA gene silencing on the proliferation of neural stem cells in rats.
METHODS: Neural stem cells of Wistar fetal rats were cultured in vitro, and were divided into six groups: blank control group, 5, 10, 15, 20 μmol/L fasudil groups, and siRNA silencing RhoA gene group. On day 3 following intervention, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the expression of RhoA mRNA and RhoA protein of neural stem cells in each group. Cellular proliferation was determined by MTT assay. The cell cycle was analyzed by flow cytometry.
RESULTS AND CONCLUSION: RhoA gene and protein expression was significantly lower in the 15, 20 μmol/L fasudil groups and siRNA silencing RhoA gene group compared with 5, 10 μmol/L fasudil groups and blank control group (P < 0.05). Cell growth speed was significantly faster compared with the 5, 10 μmol/L fasudil groups and blank control group (P< 0.05); cell number was reduced in the cell cycle G0/G1 (P < 0.05) and increased in S phase (P < 0.05). At 20 μmol/L fasudil, cell effect was not enhanced with the increased concentration. No significant difference was determined compared with 15 μmol/L fasudil group (P > 0.05). No significant difference was detected between 15, 20 μmol/L fasudil groups and siRNA silencing RhoA gene group (P > 0.05). Results suggest that Rho kinase inhibitor fasudil and RNAi-mediated RhoA gene silencing can promote the proliferation of neural stem cells in vitro. The optimal concentration of fasudil was 15 μmol/L.

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