甲泼尼龙减轻水通道蛋白4抗体对星形胶质细胞和神经元的毒性

Methylprednisolone attenuates aquaporin-4 antibody-induced cytotoxicity in astrocytes and neurons

目的探讨不同滴度水通道蛋白4(aquaporin-4,AQP4)抗体阳性血清对体外培养大鼠皮质神经细胞的毒性作用以及甲泼尼龙预处理的保护作用。方法使用原代培养的新生Wistar大鼠大脑皮质神经细胞,分别加入正常对照血清,AQP4抗体滴度1∶10、1∶320和1∶1000血清以及单独甲泼尼龙、甲波尼龙+AQP4抗体滴度1∶1000血清。培养6h后使用免疫荧光染色法观察各组星形胶质细胞、神经元的形态学改变以及数量变化。结果加入AQP4抗体阳性血清引起星形胶质细胞胞体肿胀和突起减少,神经元突起也明显减少,而甲泼尼龙预处理可减少这些形态学改变;对照组、1∶10、1∶320和1∶1000滴度血清组星形胶质细胞数目分别为(52.22±3.75)%、(45.83±4.82)%、(35.77±5.91)%和(24.68±3.40)%;神经元数目分别为(44.93±2.39)%、(39.00±4.33)%、(32.21±2.67)%和(23.35±2.65)%,各组间星形胶质细胞数量和神经元数量比较均有统计学差异(F值分别为48.09和71.41,均P<0.01),组间两两比较均有统计学差异(均P<0.01)。而甲泼尼龙+1∶1000血清组的星形胶质细胞和神经元数目分别为(44.52±3.71)%和(42.11±5.94)%,较1∶1000血清组明显增多(均P<0.01)。结论 AQP4抗体阳性血清对体外培养的大鼠星形胶质细胞和神经元有滴度依赖性细胞毒性作用,甲泼尼龙预处理能减轻其毒性。

Objective To investigate the effect of aquaporin 4 （AQP4） antibody-positive serum and the protective effect of methylprednisolone pretreatment on cultured cortical cells in rats.Methods The primary cerebral cortical cells of neonatal Wistar rat were cultured with the normal control serum,the serum with AQP4 antibody titers 1.∶ 10,1 ∶ 320,1 ∶ 1000,methylprednisolone alone,and 0.8 μmol methylprednisolone combined with 1 ∶ 1000 AQP4 antibody serum.After 6 h cultivation,morphological and quantity changes of astrocytes and neurons were oobserved by immunofluorescence staining.Results In all the AQP4 antibody positive groups,the majority of astrocytes exhibited cellular swelling,accompanied with reduced processes.The neuronal processes also presented with the similar changes.The methylprednisolone pretreatment could significantly reduce these morphological changes.The percentage of astrocytes were （52.22 ± 3.75） ％,（45.83 ± 4.82） ％,（35.77 ±5.91）％ and （24.68±3.40）％,and the percentage of neurons were （44.93±2.39）％,（39.00±4.33）％,（32.21±2.67）％ and （23.35±2.65）％ in the control group,AQP4 antibody title 1 ∶ 10,1 ∶ 320 and 1 ∶ 1000 group respectively,and there were statistical significant differences among the four groups （F=48.09 and 71.41,P＜0.01,respectively）.In the methylprednisolone±AQP4 antibody titers 1 ∶ 1000 serum group,the percentage of astrocytes and neurons [（44.52 ± 3.71）％ and （42.11 ± 5.94）％ respectively] significantly increased as compared with those of the AQP4 antibody title 1 ∶ 1000 group （P＜0.01）.Conclusions AQP4 antibody induces titer-dependent cytotoxicity on astrocytes and neurons in cultured rat cortical cells and methylprednisolone can protect against aquaporin-4 antibody-induced cytotoxicity.