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

摘要

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100.00 μg/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100.00 μg/L的TNF-α和1 g/L的环孢素,加入作用物后120 min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-α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 culture.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 methylthiazolyldiphenyl-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.