摘要
目的研究小胶质细胞上非受体型酪氨酸激酶Src通过第10号染色体缺失的磷酸酶张力蛋白同源物基因(PTEN)编码的PTEN蛋白调节细菌脂多糖(LPS)诱导神经炎症反应的分子机制。方法采用LPS诱导小胶质细胞系BV2,Western blot法检测蛋白激酶B(Akt)磷酸化水平评价PTEN活性,酶联免疫吸附法测定肿瘤坏死因子-α含量评价神经炎症反应;加入PTEN抑制剂以及Src的小干扰RNA后,观察神经炎症反应的变化,研究Src调控神经炎症机制。结果 LPS诱导BV2细胞激活后,Akt磷酸化水平显著升高(P<0.001),肿瘤坏死因子-α释放量显著增多(P<0.001),即引起明显的神经炎症反应。而抑制PTEN蛋白活性后,LPS诱导的Akt磷酸化水平进一步升高(P<0.05),炎症因子释放量进一步增多(P<0.001);干扰Src基因后,Src蛋白表达量明显下降(P<0.001),同时LPS诱导的炎症因子释放量与未干扰Src组相比明显减少,炎症反应减轻(P<0.001);PTEN和Akt磷酸化水平也明显下降(P<0.001)。结论在小胶质细胞上Src激酶可能通过调节PTEN蛋白活性调控神经炎症反应。
Objective To investigate the mechanism of non-receptor tyrosine kinase Src regulating neuroinflammation through phosphatase and tensin homology protein（PTEN） in microglia.Methods BV2 cells were incubated with PTEN inhibitor bpv（HOpic） for 2 hours,and then added with lipopolysaccharide（LPS）to induce neuroinflammation,Western blot was performed to determine the expression of phosphorylated protein kinase B（Akt） to investigate the activity of PTEN.Enzyme-linked immunosorben assay（ELISA） was used to determine the release of tumor necrosis factor α（TNF-α） to assess neuroinflammation.After PTEN inhibitor or Src specific small interfering RNA was added,the change of neuroinflammation was evaluated to study the mechanism of Src regulating neuroinflammation.Results LPS induced significant neuroinflammation in BV2 cells,as indicated by significantly increased expression of p-Akt and release of TNF-α（P〈0.001）.The PTEN inhibitor signficantly increased Akt phosphorylation（P〈0.05） and TNF-α release（P〈0.001） in LPS-induced BV2 cells compared to simply LPS-induced cells.The Src small interfering RNA significantly decreased the release of TNF-α（P〈0.001） and inhibited PTEN（P〈0.001） and Akt（P〈0.001） phosphorylation.Conclusion Srckinase may regulate neuroinflammtion response in BV2 cells by regulating the phosphorylation of PTEN.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2017年第4期534-538,共5页
Acta Academiae Medicinae Sinicae
