体外小胶质细胞活化及MRP8表达关系的研究

Relationship between microglia activation and MRP8 expression in vitro

目的建立体外小胶质细胞糖氧剥离活化模型,并探讨小胶质细胞活化与MRP8表达的关系。方法体外培养大鼠原代小胶质细胞,糖氧剥离(OGD)法建立小胶质细胞活化模型,通过形态学观察、MTT法检测不同OGD时间点小胶质细胞活力改变,实时荧光定量PCR法检测不同时间点小胶质细胞MRP8相对表达量。结果小胶质细胞OGD培养5min后细胞开始活化,活力较正常对照组明显增加(P<0.05),且在10min组细胞活力最大,OGD培养15min、30min、60min后细胞活力明显下降(P<0.05)。MRP8在小胶质细胞中表达随着OGD时间延长先增加后降低,在10min组达高峰(P<0.05)。结论首次在体外证实活化的小胶质细胞表达MRP8增加,这可能是其参与多种中枢神经系统疾病发病的机制之一。

Objective To establish microglial activation model in vitro by glucose-oxygen deprivation and explore the relationship between microglia activation and expression of MRP8. Methods First prepared primary cultured rat micro- glial cells. Microglial activation model was established by oxygen-glucose deprivation （OGD） mehtod. Using morphological observation, microglia viability at different OGD time points was tested by MTT assay. Real time PCR method was used to detect the relative expression of MRP8 at different time points. Results Microglia cells began to activate and the viability of microglia cells increased obviously（ P 〈 0.05） after 5rain of OGD, was the highest at 10rain. Then decreased significantly after longer times （15,30, 60rain） of OGD（P 〈 0.05）. The expression of MRP8 in microglia cells increased at first to reach the peak at 10rain （ P 〈 0.05 ）, then decreased as the OGD time extended. Conclusion We firstly confirmed that ac- tivated microglia expressed higher MRP8 in vitro. This may be an important pathogenesis for microglia cells participating in many central nervous system disease development.