GSK-3β活性与小胶质细胞TLR4受体在POCD发生中的作用机制  

Role of GSK-3β activity and microglial TLR4 receptor in POCD

作　　者：张波[1] 吴世政[1] 胡全忠[1] 侯倩[1] 才鼎[1] 钱燕宁[2] 

机构地区：[1]青海省人民医院神经内科,青海西宁810007 [2]南京医科大学,江苏南京210029

出　　处：《中国生化药物杂志》2015年第4期39-41,共3页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81070276)

摘　　要：目的体外实验中检测小胶质细胞糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)的活性和Toll样受体4(Toll-like receptor 4,TLR4)蛋白的表达,探究手术后认知障碍(post operative cognitive dysfunction,POCD)的可能作用机制。方法原代培养小胶质细胞,倒置显微镜观察细胞形态,采用神经胶质酸性蛋白(glial fibrillary acidic protein,GFAP)免疫荧光法鉴定小胶质细胞;采用脂多糖(lipopolysaccharides,LPS)诱导小胶质细胞损伤,筛选最佳POCD造模条件;采用MTT法检测小胶质细胞活力;采用Western blot方法检测小胶质细胞中GSK-3β和TLR4蛋白的表达。结果 GFAP免疫荧光鉴定结果显示阳性,大鼠小胶质细胞的原代培养成功;MTT结果表明,最佳PODC造模条件为LPS(100 ng/mL)诱导小胶质细胞损伤7h;Western blot结果表明,与空白对照组相比,LPS可显著性地上调小胶质细胞内GSK-3β和TLR4蛋白的表达(P<0.01)。结论 PODC的发病机制可能与LPS上调小胶质细胞中GSK-3β和TLR4蛋白的表达有关。Objective To explore glycogen synthase kinase -3β（ GSK-3β） activity and Toll-like receptor 4 （ TLR4 ） proteins expression of microglia were tested in vitro experiments, and the possible mechanism of postoperative cognitive dysfunction（POCD）.Methods The cell morphology of primary culture microglia was observed by inverted microscope;microglia were identified by glial fibrillary acidic protein （ GFAP ） immunofluorescence;the best POCD modeling conditions of microglia injury induced by lipopolysaccharides（ LPS） were screened ; microglia vigor was assayed by MTT ; the proteins expressions of GSK-3βand TLR4 of microglia were detected by Western blot.Results GFAP immunofluorescence showed a positive result that primary culture of rat microglia was successful;MTT result showed that the best PODC modeling conditions of microglia injury induced by LPS （100 ng/mL） was 7h; Western blot results showed that the preotein expressions of GSK-3βand TLR4 of microglial cells were up-regulated by LPS compared with the control group,and there were significantly differences （P〈0.01）.Conclusion PODC pathogenesis may be associated with LPS that could up-regulat the protein expression of GSK-3βand TLR4 in microglial cells.

关 键 词：术后认知障碍 糖原合成酶激酶-3Β TOLL样受体4 小胶质细胞 作用机制 

分 类 号：R392[医药卫生—免疫学]