APP5肽类似物P165和罗格列酮对SH-SY5Y细胞IRS-1和p-CREB表达的影响

目的观察APP5肽类似物P165和罗格列酮(rosiglitazone)对链脲佐菌素(streptozotocin,STZ)损伤人神经母细胞瘤株SH-SY5Y细胞后,胰岛素信号通路相关因子IRS-1和p-CREB表达水平的影响。方法将人神经母细胞瘤株SH-SY5Y细胞分为对照组、STZ损伤模型组(STZ 0.8 mmol/L)、罗格列酮保护组(STZ 0.8 mmol/L+罗格列酮20μmol/L),APP5肽类似物P165保护组(STZ 0.8 mmol/L+P165 30μmol/L),进行细胞形态观察,IRS-1和p-CREB免疫细胞荧光染色观察,采用image-pro plus图像分析软件测定平均吸光度值。结果与对照组相比,STZ损伤组细胞生长分裂减慢、突起缩短,细胞胞体皱缩,IRS-1和p-CREB免疫荧光染色强度减弱;与STZ损伤组相比,罗格列酮组和P165组突起伸展,细胞形态和细胞胞体皱缩现象明显改善,细胞出现强荧光染色。平均吸光度值测定结果显示:与对照组相比,STZ损伤组IRS-1和p-CREB平均吸光度值降低(P<0.008),与STZ损伤组相比,P165组和罗格列酮组平均吸光度值有所上升,差异均有统计学意义(P<0.008)。结论 APP5肽类似物P165和罗格列酮可能通过增加IRS-1、p-CREB的表达来改善STZ对于SH-SY5Y细胞的损伤,从而达到保护神经元的作用。

Effects of P165 and rosiglitazone on the expression of IRS-1 and p-CREB in SH-SY5Y cell

Objective To investigate the effects of P165(the analog of APP5 peptide) and rosiglitazone on the expression of insulin signaling pathway related factors IRS-1 and p-CREB in SH-SY5Y cells after streptozotocin(STZ)-induced injury. Methods Human neuroblastoma cell line SH-SY5Y cells were divided into four groups: control group, STZ injury group(STZ 0.8 mmol/L), rosiglitazone protection group(STZ 0.8 mmol/L+rosiglitazone 20 μmol/L) and P165 protection group(STZ 0.8 mmol/L+P165 30 μmol/L), and were observed for morphology of cells and the immunofluorescence staining of IRS-1 and p-CREB, and we determined mean optical density using Image-pro plus software. Results Compared with the control group, cells of STZ injury group grew and divided slower, cell processes were shorter and the cell body shrunk, the intensity of immunofluorescence staining of IRS-1 and p-CREB were reduced. Compared with STZ injury group, the cells in two protection groups had neurite extension and ameliorated in cell body shrinkage and morphology, and showed stronger immunofluorescence staining. The mean optical density of IRS-1 and p-CREB in STZ injury group decreased than that of the control group(P<0.008), and the density in two protection groups increased compared with STZ injury group (P<0.008). Conclusion P165 and rosiglitazone could ameliorate the STZ-induced injury to SH-SY5Y by improving the expression of IRS-1 and p-CREB, and thereby protect the neurons.