CAJ | 학술논문

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。
목적：체외실험중검측소효질세포당원합성매격매-3β（glycogen synthase kinase-3β，GSK-3β）적활성화Toll양수체4（Toll-like receptor 4，TLR4）단백적표체，탐구수술후인지장애（post operative cognitive dysfunction ，POCD）적가능작용궤제。방법원대배양소효질세포，도치현미경관찰세포형태，채용신경효질산성단백（ glial fibrillary acidic protein，GFAP）면역형광법감정소효질세포；채용지다당（ lipopolysaccharides，LPS）유도소효질세포손상，사선최가POCD조모조건；채용MTT법검측소효질세포활력；채용Western blot방법검측소효질세포중GSK-3β화TLR4단백적표체。결과 GFAP면역형광감정결과현시양성，대서소효질세포적원대배양성공；MTT결과표명，최가PODC조모조건위LPS（100 ng／mL）유도소효질세포손상7h；Western blot결과표명，여공백대조조상비，LPS가현저성지상조소효질세포내GSK-3β화TLR4단백적표체（P＜0.01）。결론 PODC적발병궤제가능여LPS상조소효질세포중GSK-3β화TLR4단백적표체유관。
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.