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Effect of astragalus polysaccharides on expression of MMP2 and MMP9 in annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats
Authors:YANG Bin  HUANG Jun-qing  ZHANG Ji-wei
Institution:The Second Department of Osteopathy, Henan Province Hospital of TCM, Zhengzhou 450000, China
Abstract:AIM: To investigate the effect of astragalus polysaccharides (AP) on the expression of matrix metalloproteinase 2 (MMP2) and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats. METHODS: The model rats were randomly divided into model group (M group), and low-dose and high-dose AP treatment groups (L-AP and H-AP groups). The rats in sham operation group were used as negative control group (NC group). In addition, all the annulus fibrosus tissues were used for primary cell culture. Histological analysis was performed using HE staining and Safranin O staining. The expression of MMP2, MMP9, tissue inhibitor of metalloproteinase 2 (TIMP2) and collagen Ⅳ at mRNA and protein levels was analyzed by immunohistochemistry, Western blot and RT-qPCR. Cell-collagen adhesion assay was used to detect annulus fibrosus cell-collagen adhesion. RESULTS: The intervertebral discs of M group were degenerated, while astragalus polysaccharide improved the degenerative disc disease in the rats with cervical spondylosis. Compared with NC group, the expression of MMP2 and MMP9 in the annulus fibrosus tissues of M group increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression of MMP2 and MMP9 in L-AP group and H-AP group was significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). The cell-collagen adhesion in M group was significantly lower than that in NC group (P<0.05). Compared with M group, the cell-collagen adhesion in L-AP group and H-AP group was increased significantly (P<0.05). Compared with NC group, the expression of MMP2 and MMP9 in annulus fibrosus cells of M group was increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression levels of MMP2 and MMP9 in L-AP group and H-AP group of fibrocytes were significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). CONCLUSION: Astragalus polysaccharides inhibit the expression of MMP2 and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats and regulate the dynamic balance of MMPs and TIMPs in the extracellular matrix, thus inhibiting the degradation of collagen in the intervertebral disc matrix and having the potential research value for the treatment of intervertebral disc degeneration.
Keywords:Astragalus polysaccharides  Cervical spondylosis  Intervertebral disc  Annulus fibrosus  Matrix metalloproteinases  Extracellular matrix  
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