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| 抗血小板溶栓素对缺糖缺氧再灌注损伤大鼠原代皮质神经细胞保护作用 |
| Effect of anti-platelet thrombolysin on rat primary cortical neurons during oxygen and glucose deprivation/reperfusion injury |
| 投稿时间：2015-10-12 |
| DOI：10.3969/j.issn.1000-0399.2016.03.001 |
| 中文关键词: 抗血小板溶栓素 神经细胞 Toll样受体4 Rho蛋白A Rho相关卷曲螺旋形成蛋白激酶1/2 |
| 英文关键词: Anti-platelet thrombolysin Neuron Toll like receptor 4 RhoA Rho associated coiled coil forming protein kinase 1/2 |
| 基金项目:安徽省自然科学基金资助项目(项目编号:1208085QH140) |
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| 中文摘要: |
| 目的 探讨抗血小板溶栓素(APT)对原培养大鼠皮质神经细胞缺糖缺氧损伤的保护作用及其机制。方法 大鼠原代皮质神经细胞培养6 d后,经APT 80μg/mL、40μg/mL、20μg/mL、Toll样受体4(TLR4)阻断剂HTA12510μg/mL预处理24 h,皮质神经细胞缺糖缺氧3 h,恢复正常培养24 h,建立缺糖缺氧再灌注损伤模型。实验分为正常组,模型组,APT低、中、高组,HTA组。倒置显微镜下观察细胞形态,免疫组化染色鉴定神经元。MTT法检测细胞存活率。比色法检测细胞培养液中丙二醛(MDA)含量和乳酸脱氢酶(LDH)活性。G-LISA法检测细胞中Rho蛋白A(RhoA)活性,Western-bloting检测细胞中TLR4、Rho相关卷曲螺旋形成蛋白激酶(ROCK)蛋白表达水平。结果 细胞培养6 d后,免疫组化鉴定显示神经细胞纯度达90%以上;与模型组比较,APT低、中、高剂量能明显增加神经细胞存活率;中、高剂量能明显降低培养液中MDA含量和LDH活性,降低细胞中RhoA活性,抑制细胞中TLR4,ROCK1/2蛋白表达。结论 APT对大鼠原代皮质神经细胞缺糖缺氧再灌注损伤有较好的保护作用,其机制与抑制TLR4、RhoA、ROCK信号通路有关。 |
| 英文摘要: |
| Objective To study the effect of anti-platelet thrombolysin(APT) on rat primary cortical neurons during oxygen and glucose deprivation/reperfusion injury(OGD/Rep) and the underlying mechanism.Methods Rat primary cortical neurons were cultured with oxygen and glucose deprivation for 180 minutes and returned to normal culture for 24 hours. Cells were divided into six groups including the normal group, model group, HTA125(Toll-like receptor 4 blocker) 10μg/mL group, APT 20μg/mL, 40μg/mL, 80μg/mL group. Cell viability was detected by MTT assay. MDA level and LDH activity were detected by chemical colorimetry. RhoA activity was determined using absorbance based G-LISA RhoA activation assay. The expressions of TLR4, ROCK1/2 were detected by Western-blot.Results Compared with model group, APT significantly reduced MDA level, LDH activity and neuronal cell viability, and reduced the RhoA activity, the expressions of TLR4 and ROCK1/2.Conclusion APT has protective effect on rat primary cortical neurons during OGD/Rep, which may be associated with the inhibition of TLR4/RhoA/ROCK pathway. |
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