Effect of consistence pressure on expression of Wnt/beta-catenin signaling pathway and VEGF of the rabbit intervertebral disc motion segments vertebral endplate in vitro organ culture
ZHAN Jiawen1,3 FENG Minshan2,3 WANG Shangquan1▲ ZHU Liguo2,3 HAN Tao1 YIN Xunlu1 WEI Xu4
1.Department of the General Orthopedics, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China;
2.the Second Department of Spine, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China;
3.the Key Laboratory of Chinese Medicine Technology in Beijing, Beijing 100102, China;
4.Research Offices, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China
Abstract：Objective To study the effect of consistence pressure on the expression of VEGF and beta -catenin in vertebral endplate, and further explore the molecular mechanism between VEGF and intervertebral disc degeneration. Methods A total of 16 New Zealand rabbits were sacrificed after removed under sterile conditions in the spinal motion segment, were randomly divided into control group and pressure group(8 rabbits in each group). Rabbits were placed in vitro loading and culture device for culture, in which the pressure group were treated with consistence pressure 3 kg, the control group did not under pressure, pro-culture and 3, 7, 14 d after culture, 10 samples were collected from each group, the expression of VEGF and beta-catenin were detected by immunohistochemistry (IHC), Western blot and Real time-PCR. Results IHC staining showed that the VEGF staining intensity in the endplate under pressure 7 d after cultured decreased significantly(0.182±0.063), compared with the control group(0.237±0.051), the difference was statistically significant (P < 0.05); the staining intensity was further decreased at 14 d after culture (0.156±0.035), compared with the control group (0.211±0.038), the difference was statistically significant(P < 0.05). On the contrary, the consistence pressure increased the expression of beta-catenin in the endplate, and compared with that of the control group, the difference was statistically significant (P < 0.05). Western blot results showed that compared with fresh specimen and control group, consistence pressure significantly increased the expression of beta-catenin, the differences were statistically significant (P < 0.05). Real-time PCR results showed the relative expression of VEGF gene was reduced by consistence pressure(0.842±0.002), and the difference was significant compared with the control group (0.965±0.005, P < 0.05). Conclusion Consistence pressure leads to a decrease of VEGF expression and an increase of the expression of beta-catenin in the vertebral endplate of spinal motion segment, which suggests that continuous pressure may regulate the expression of VEGF in the vertebral endplate through the Wnt/beta-catenin signaling pathway.
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2018, Vol. 15 
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