Abstract：Objective To develop an in vitro organ culture model of rabbit intervertebral disc, and cyclic mechanic pressure was applied based on this model, and to investigate the change of ANK. To explore the regulatory effect of exogenous TGF-β1 on the expression of ANK. Methods The New Zealand white rabbits of 6 months old were randomly divided into cyclic mechanical pressure group and control group. Rabbits intervertebral disc were taken out completely with part of the vertebrae from lumbar spines by asepsis. The whole rabbit intervertebral discs were cultured in medium with 20% fetal bovine serum supplemented. And cyclic mechanical pressure at 0.2 MPa was applied for cyclic mechanical pressure group different periods, once every 30 minutes in a day. The change of histomorphology, the cell viability and the protein expression levels of progressive ankylosis gene within intervertebral disc tissue were assessed by HE staining, NBT-DAPI,Western blotting at 0 day,7 days and 14 days. The protein expression levels of progressive ankylosis gene within intervertebral disc were cultured at cyclic mechanical pressure for 7 days after exogenous TGF-β1 stimulation assessed by Western blotting. Results Compared with 0 day group, there were minimal changes in histomorphology, cell viability and the protein expression levels of progressive ankylosis gene at control group for 7 days(P＞0.05). There were minimal changes in histomorphology at cyclic mechanical pressure for 7 days. There were obvious change of damage in histomorphology both control group and cyclic mechanical pressure group for 14 days(P＜0.05). The cell viability was decreased at 7, 14 days of cyclic mechanical pressure group and 14 days of control group(P＜0.05). The expression levels of progressive ankylosis gene were significantly decreased at 7,14 days of cyclic mechanical pressure group and 14 days of control group(P＜0.05). On the other hand, after exogenous TGF-β1 stimulation, the expression level of progressive ankylosis gene was significantly increased at 7 days of cyclic mechanical pressure group(P＜0.05). Conclusions Based on the model of rabbit intervertebral disc, cyclic mechanical pressure could induce significant decrease of the expression level of progressive ankylosis gene. It may be closely related with the occurrence and development of intervertebral disc degeneration. And it confirms that exogenous TGF-β1 has a positive regulatory role for the expression level of progressive ankylosis gene to protect intervertebral disc from degeneration.
徐宏光 章平治 宋俊兴 熊寿良 吕坤 钟民 张梦莹 所起凤. 循环机械压力诱导下兔椎间盘器官模型ANK蛋白的表达[J]. 中华解剖与临床杂志, 2014, 19(1): 51-57.
Xu Hongguang, Zhang Pingzhi, Song Junxing, Xiong Shouliang, Lyu Ku, Zhong Min, Zhang Mengying, Suo Qifeng. ANK expression in rabbit organ model of intervertebral disc induced by cyclic mechanic pressure. Chinese Journal of Anatomy and Clinics, 2014, 19(1): 51-57.
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2014, Vol. 19 
