神经钙调蛋白在肿瘤坏死因子α诱导的星形胶质细胞损伤中的作用

Effect of calcineurin on astrocyte injury induced by tumor necrosis factor-α

目的探讨神经钙调蛋白在肿瘤坏死因子α（TNF—α）诱导的星形胶质细胞损伤中的作用。方法以1日龄的新生Wistar大鼠为实验动物，取大脑皮层脑组织培养星形胶质细胞，将培养的细胞完全随机分为对照组、TNF-α作用组、环孢素（神经钙调蛋白抑制剂）单纯作用组、环孢素和TNF-α联合作用组。对照组不加入TNF-α，TNF-α作用组加入100．00μg／L的TNF-α，环孢素单纯作用组加入1g／L的环孢素，环孢素和TNF-α联合作用组加入100．00μg／L的TNF-α和1g／L的环孢素，加入作用物后120min采用噻唑蓝比色法比较4组吸光度值、乳酸脱氢酶（LDH）释放率及细胞凋亡和坏死情况。结果TNF-α作用组星形胶质细胞吸光度值明显低于对照组（0．65±0．21比0．93±0．13），差异有统计学意义（P〈0．05），但环孢素和TNF-α联合作用组吸光度值与TNF-α作用组比较差异无统计学意义（P〉0．05）；TNF—α作用组LDH释放率明显高于对照组[（38．5±0．8）％比（20．3±1．1）％]，差异有统计学意义（P〈0．01），但环孢素和TNF．仅联合作用组LDH释放率与TNF-α作用组比较，差异无统计学意义（P〉0．05）。TNF-α作用组细胞凋亡率和坏死率均明显高于对照组[（17．4±1．8）％比（7．8±1．2）％，（35．7±3．9）％比（5．2±0．7）％]，但环孢素和TNF-α联合作用组细胞凋亡率和坏死率与TNF-α作用组比较，差异无统计学意义（P〉0．05）。结论TNF-α引起的星形胶质细胞损伤是通过非神经钙调蛋白途径实现的。

Objective To explore the effect of calcineurin on astrocyte injury induced by tumor necrosis factor-α（TNF-α）. Methods Neonatal Wistar rats born within 24 h were as experimental subjects; cerebral cortex tissues were sampled for astrocyte cuhure. Primary cultured astrocytes were randomly divided into control group, TNF-α group（ adding TNF-α 100. 00 μg/L）, ciclosporin A（ adding CsA） group（ CsA 1 g/L） and TNF-α ＋ CsA group（ adding TNF-α 100. 00 μg/L and CsA 1 g/L）. One hundred and twenty minutes after adding reagents, absorbance value, lactate dehydrogenase （LDH） release rate, apoptosis rate, necrosis rate were tested by methylth- iazolyldiphenyl-tetrazolium bromide assay. Results In TNF-α group, absorbance value was significantly lower than that in control group（0. 65 ± 0. 21 vs 0. 93 ± 0. 13 ） （ P 〈 0. 05 ） ; LDH release rate was significantly higher than that in control group [ （ 38.5 ± 0. 8 ） % vs （ 20. 3 ± 1.1 ） % ] （ P 〈 0. 01 ） ; apoptosis rate and necrosis rate were significantly higher than those in control group [ （ 17.4 ± 1.8 ） % vs （7.8 ± 1.2） %, （ 35.7 ± 3.9 ） % vs （ 5.2 ± 0. 7 ） % ] （P 〈 0. 05）. Differences of absorbance value, LDH release rate, apoptosis rate and necrosis rate between TNF-α group and TNF-α ＋ CsA group were all not significant （ P 〉 0. 05 ）. Conclusion TNF-α induced astrocyte injury is achieved through non-calcineurin pathway.