BACKGROUND: Foreign studies have shown that hyaluronan can contribute to the improvement of motor function after spinal cord injury in rats, but its mechanism is unclear.rnOBJECTIVE: To study the effect of hyaluronan on expression of neurotrophic factors in astrocytes.rnMETHODS: Cultured astrocytes were obtained from the brain of SD rats under sterile conditions. Passage 3 cells were identified by immunochemistry method. Passage 4 cells were treated with hyaluronan (Mr: 125 000-175 000) for 24 hours, and control group was set.rnRESULTS AND CONCLUSION: The results of immunofluoresence showed that the positive rate of primary astrocytes was nearly 100%. The results of Western blotting and reverse transcription-PCR showed that expressions of brain-derived neurotrophic factor and vascular endothelial growth factor were significantly increased in 100 mg/L and 1 000 mg/L groups compared with the control, 1 mg/L and 10 mg/L groups (P < 0.05). Double-labeling immunofluoresence also showed that, under the effect of 1 000 mg/L hyaluronan, the expressions of brain-derived neurotrophic factor and vascular endothelial growth factor in astrocytes were strongly increased compared with the control group. It is confirmed that small molecular-weight hyaluronan can induce astrocytes to up-regulate expressions of brain-derived neurotrophic factor and vascular endothelial growth factor.%背景:国外研究表明透明质酸可以促进脊髓损伤后大鼠运动功能的改善,但其机制尚不清楚.目的:观察透明质酸对星形胶质细胞营养因子表达的影响.方法:无菌条件下分离SD大鼠脑组织星形胶质细胞进行原代培养,对培养第3代细胞采用免疫化学方法对其鉴定.对第4代细胞给予透明质酸(相对分子质量125 000~175 000)作用星形胶质细胞24 h,并设置对照组.结果与结论:免疫化学方法鉴定原代培养星形胶质细胞阳性率近100%.Western blot和RT-PCR分析结果显示,胶质细胞在质量浓度100,1 000 mg/L 透明质酸作用下同10,1 mg/L透明质酸组和对照组比较,脑源性神经营养因子和血管内皮生长因子表达明显升高(P < 0.05);免疫荧光双标显示在质量浓度1 000 mg/L 透明质酸作用下,同对照组相比,胶质细胞脑源性神经营养因子和血管内皮生长因子表达明显增加.证实小分子量透明质酸可上调星形胶质细胞中脑源性神经营养因子和血管内皮生长因子的表达.
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