Abstract. The aim of the present study was to assess miR-625 and Fas expression in normal and degenerative cervical cartilage endplate (CEP) tissues. Following biof-informatics analysis, the Fas gene was predicted to be one of the targets of miR-625. Quantitative PCR (qPCR) and western blotting were used to detect miR-625 and Fas expression in normal and degenerative CEP. A luciferase reporter assay was used to identify whether miR-625 could directly target the 3' untranslated region (3'-UTR) of Fas. Lentiviral overexpression and/or inhibition vectors of miR-625 (pre-miR-625)/antigomiR-625 were constructed to determine whether overexpression or inhibition of miR-625 could affect Fas and B-cell lymphoma 2 (Bcl-2) expression in cartilaginous endplate cells (CECs) and tissues. qPCR analysis demonstrated that miR-625 expression in degenerative CEP was significantly lower than in normal CEP tissue, while the production of Fas in degenerated CEP was significantly higher. Results from western blotting also showed a significant increase in Fas expression in degenerative CEP. miR-625 can bind directly to the 3'-UTR of the Fas gene. However, this inhibition was attenuated by a target mutation in the miR-625-binding site of the 3'-UTR of Fas mRNA. In addition, following transfection of CECs with pre-miR-625 and antigomiR-625, expression of Fas significantly decreased and increased, respectively, and Bcl-2 expression was upregulated and downregulated, respectively. Upregulation of miR-625 can inhibit Fas expression and further affect Bcl-2 expression in CEP degeneration, suggesting that miR-625-mediated inhibition of the Fas gene is important in cervical degeneration.
Proanthocyanidins were extracted and purified from grape seeds, and the protective effect and mechanism of proanthocyanidins on spinal cord injury were explored. After extraction optimization and purification, the proanthocyanidins with purity of 92.53% were obtained. The rats were divided into control, model and treatment groups. The spinal cord injury model was established in model and treatment groups. Then, the treatment group was treated with 40 mg/kg proanthocyanidins. On the 1st, 3rd and 7th day after modeling, compared with model group, in model group the Basso-Beattie-Bresnahan scores were significantly decreased (p < 0.05). On the 7th day after modeling, compared with model group, in treatment group the spinal cord tissue superoxide dismutaseand glutathione peroxidase levels were significantly increased (p < 0.05), the malondialdehyde level was significantly decreased (p < 0.05), the necrosis factor α and interleukin 1βlevels were significantly decreased (p < 0.05), and the interleukin 10 level was significantly increased (p < 0.05). In conclusion, proanthocyanidins have protective effect on spinal cord injury in rats. The mechanisms may be related to its reducing oxidative stress and inflammation in spinal cord tissue.
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