活性多肽GRGDS对氧糖剥夺诱导PC12细胞损伤的保护作用及其机制研究

Protective effect and mechanism of active peptide GRGDS on PC12 cells damage by oxygen-glucose deprivation

目的研究活性多肽GRGDS对氧糖剥夺(OGD)损伤模型中大鼠神经细胞(PC12细胞)的保护作用,并探讨其作用机制。方法将PC12细胞分成对照组、ODG组、活性多肽GRGDS治疗组,通过氧糖剥夺模拟体外脑缺血建立损伤模型。用噻唑蓝(MTT)比色法和流式细胞仪检测氧糖剥夺后细胞凋亡情况,酶联免疫吸附试验法(ELISA)检测氧糖剥夺后PC12细胞上清液中炎症因子,肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)的含量变化,蛋白质印迹法(Western blot)检测凋亡通路相关蛋白表达情况。结果MTT法和流式细胞仪检测结果表明,活性多肽GRGDS显著降低氧糖剥夺后PC12细胞凋亡(P<0.05);ELISA检测结果表明,活性多肽GRGDS明显降低氧糖剥夺后PC12细胞上清液中TNF-α和IL-1β的含量(P<0.05),Western blot结果显示,活性多肽GRGDS可显著降低氧糖剥夺损伤介导的PC12细胞p-JNK、Bax、cleaved caspase 3等的蛋白表达水平(P<0.01)。结论活性多肽GRGDS对氧糖剥夺损伤的PC12细胞具有保护作用,其机制可能与抗凋亡、抗炎作用有关。

Objective To study the protective effect of active peptide GRGDS on rat nerve cells(PC12 cells)in oxygen glucose deprivation(OGD)injury model and explore its mechanism of action.Methods PC12 cells were divided into control group,ODG group,and active peptide GRGDS treatment group.The injury model was established by simulating in vitro cerebral ischemia by oxygen and sugar deprivation.MTT and flow cytometry were used to detect apoptosis after oxygen-glucose deprivation.ELISA method was used to detect the changes of inflammatory factors TNF-αand IL-1βin PC12 cell supernatant after oxygen-glucose deprivation.Western blot was used to detect the expression of apoptosis pathway-related proteins.Results The results of MTT and flow cytometry showed that the active peptide GRGDS significantly reduced the apoptosis of PC12 cells after oxygen glucose deprivation(P<0.05).ELISA test results showed that the active peptide GRGDS significantly reduced the content of TNF-αand IL-1βin the supernatant of PC12 cells after oxygen-glucose deprivation.(P<0.05).Western blot results showed that the active peptide GRGDS significantly reduced the expression levels of p-JNK,Bax,and cleaved caspase 3 in PC12 cells mediated by oxygen-glucose deprivation injury(P<0.01).Conclusion The active peptide GRGDS has protective effect on PC12 cells damaged by oxygen and glucose deprivation.The mechanism may be related to anti-apoptotic and anti-inflammatory effects.