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脑卒中是人类疾病中最常见的脑血管疾病,已经成为人类死亡的第二大原因[1]。脑卒中引起的一系列并发症和后遗症给患者家庭带来了不可估量的负担。脑卒中分为缺血性脑卒中和出血性脑卒中两种,其中,缺血性脑卒中又称脑中风,是脑卒中主要的发病方式,约占脑卒中患者的83%以上[2]。目前,尚无有效的治疗药物用于脑卒中引起的损伤,尤其是对于神经损伤的治疗[3]。活性多肽GRGDS是由甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸(Gly-Arg-Gly-Asp-Ser,GRGDS)5种氨基酸构成,主要通过形成一个β转角的方式与其他细胞发生黏连[4],因而,能够阻断细胞外基质和细胞表面整合素的结合和黏附,可应用于组织工程或者癌症和肿瘤方面。本试验采用PC12细胞体外模拟脑缺血模型,探讨活性多肽GRGDS对氧糖剥夺损伤后的PC12细胞是否具有保护作用。
Protective effect and mechanism of active peptide GRGDS on PC12 cells damage by oxygen-glucose deprivation
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摘要:
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。 Abstract: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-α and IL-1β in 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-α and IL-1β in 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. -
Key words:
- active peptide /
- GRGDS /
- oxygen glucose deprivation /
- PC12 cells /
- apoptosis
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