氧糖剥夺条件下PC12细胞RAGE的表达及对其损伤的影响

目的探讨大鼠肾上腺嗜铬细胞瘤(PC12)细胞在氧糖剥夺(OGD)的条件下,晚期糖基化终产物受体(RAGE)的表达及其对PC12细胞损伤的影响。方法PC12细胞随机分为3组:OGD培养组(PC12细胞在无血清无糖的DMEM培养液中厌氧培养);封闭RAGE的OGD培养组(PC12细胞在加5μg/mlRAGE抗体的无血清无糖DMEM培养液中厌氧培养);对照组(PC12细胞在无血清无糖DMEM培养液中培养)。免疫组化法检测RAGE表达。收集细胞上清液,检测乳酸脱氢酶(LDH)活性和一氧化氮(NO)含量,并对细胞死亡率进行检测。结果在OGD条件下PC12细胞培养8、11、20h均有RAGE表达,与对照组相比表达均明显增加,OGD培养组与封闭RAGEOGD培养组相比,LDH活性差异显著。随着厌氧时间的延长,PC12细胞死亡率明显增高,封闭RAGEOGD培养组与OGD培养组相比,PC12细胞死亡率明显降低。NO含量差异无显著意义,但与对照组相比差异显著。结论在OGD条件下,PC12细胞RAGE表达增高。RAGE的表达对细胞的损伤起促进作用。

Expression of Receptor for Advanced Glycation Endproduct in Pheoehromocytoma PC12 Cells and Its Effect on Cell Injury under Condition of Oxygen-Glucose Deprivation

Objective To explore the expression of receptor for advanced glycation endproduct(RAGE)in rat pheoehromocytoma PC12 and its effect on the injury of PC12 cells under the condition of oxygen-glucose deprivation(OGD).Methods Divide PC12 cells into 3 groups randomly.The cells in group 1 was cultured under OGD condition in serum-free and carbohydrate-free DMEM,and those in group 2 under OGD condition in serum-free and carbohydrate-free DMEM with addition of antibody against RAGE,while those in group 3 were culture in serum-free and carbohydrate-free DMEM.Determine the expression of RAGE by immunohistochemical staining.Collect the cell supernatant and test for lactate dehydrogenase(LDH)activity,nitrogen monoxide content and cell death rate.Results The expression of RAGE were proved in PC12 cells in group 1 after culture for 8,11 and 20 h,and the expression levels were significantly higher than those in group 3.The LDH activity of PC12 cells in group 1 cultured was significantly higher than that in group 2.The death rate of PC12 cells increased with the increasing hours for culture under OGD condition,and was significantly higher in group 1 than that in group 2.The nitrogen monoxide content showed no significant difference in the PC12 cells in groups 1 and 2,but was significantly higher in the 2 groups than that in group 3.Conclusion The expression of RAGE in PC12 cells increased under OGD condition and promoted the injury of cells.