摘要
目的探讨头孢曲松钠在水通道蛋白4(AQP4)抗体诱导的星形胶质细胞损伤中的作用以及机制。方法常规体外培养新生SD大鼠大脑皮质细胞,将培养的细胞分为4组,分别加入健康人血清(对照组)、AQP4抗体阳性患者血清、头孢曲松钠+AQP4抗体阳性血清以及单纯头孢曲松钠。细胞培养24h后采用免疫组织化荧光染色观察不同组星形胶质细胞数目的变化,采用比色法测定上清液谷氨酸浓度以及免疫印迹分析谷氨酸转运体-1(GLT-1)蛋白表达水平。结果和对照组比较,AQP4抗体阳性血清组星形胶质细胞数目和谷氨酸转运体-1(GLT-1)蛋白表达明显减少,上清液谷氨酸浓度明显增高(均P<0.01),而单纯头孢曲松钠组仅显著增加GLT-1蛋白表达(P<0.01),并不影响星形胶质细胞数目和谷氨酸水平(P>0.05);和AQP4抗体阳性组比较,头孢曲松钠+AQP4抗体阳性血清组星形胶质细胞数目和GLT-1蛋白表达增加,上清液谷氨酸浓度明显下降(P<0.01)。结论头孢曲松钠可能通过上调星形胶质细胞GLT-1表达、减少细胞外液谷氨酸水平发挥减轻AQP4抗体对体外培养的大鼠脑皮质星形胶质细胞的毒性作用。
Objective To observe the effects of ceftriaxone on toxicity of water channel protein-4 （aquaporin 4, AQP4） antibody to astrocytes and the possible mechanisms. Methods The routine methods were used to extract cortical cells from 1-day old Sprague-Dawley rats, the cultured cerebral cortical cells were treated with healthy sera, AQP4 antibody positive sera, ceftriaxone -- AQP4 antibody positive sera, ceftriaxone respectively. After 24 h cultivation, the number of astrocytes was observed by immunofluoreseence staining. The glutamate concentration in the supernatants was measured by colorimetry, and the expression of glial glutamate transporter-1 （GLT-1） was evaluated by western blot. Results The number of astroeytes and expression of GLT 1 in astroeytes were significantly decreased and the glutamate concentration was significantly increased in the AQP4 antibody positive group compared with the control group （P〈0.01）. Ceftriaxone increased the expression of GLT 1 （P〈0.01）, and did not change the number of astrocytes and the glutamate concentration comparing the ceftriaxone only group with the control group （P〉0.05 ）. The number of astrocytes and expression of GLT-1 in astrocytes were significantly increased and the glutamate concentration was significantly decreased in the ceftriaxone＋ AQP4 antibody positive group compared with the AQP4 antibody positive group （P 〈0.01）. Conclusions Ceftriaxone could attenuates the eytotoxic effect of anti-aquaporin-4 antibody on cultured rat cortical astrocytes by upregulating the expression of GLT-1 in astrocytes and decreasing the glutamate concentration in the extraeellular fluid.
出处
《中国神经免疫学和神经病学杂志》
CAS
2015年第3期162-165,共4页
Chinese Journal of Neuroimmunology and Neurology
