扇贝裙边糖胺聚糖对SH-SY5Y细胞氧化及凋亡影响  

EFFECT OF SCALLOP SKIRT-GLYCOSAMINOGLYCAN ON OXIDATION AND APOPTOSIS IN SH-SY5Y CELLS

作　　者：赵小丹[1] 鞠传霞[2] 侯琳[1] 刘存[1] 宋金莲[3] 

机构地区：[1]青岛大学医学院生物化学与分子生物学教研室,山东青岛266021 [2]青岛大学医学院药理学教研室 [3]青岛市妇女儿童医疗保健中心检验科

出　　处：《齐鲁医学杂志》2014年第5期427-429,432,共4页Medical Journal of Qilu

基　　金：山东省自然科学基金资助项目(ZR2013HQ010);山东省医药卫生科技发展计划项目(2013WS0256);青岛市民生计划项目(13-1-3-48-nsh)

摘　　要：目的探讨扇贝裙边糖胺聚糖（SS-GAG）对6-羟基多巴胺（6-OHDA）诱导的SH-SY5Y细胞丙二醛（MDA）、乳酸脱氢酶（LDH）含量及凋亡相关基因Bax、Bcl-2、Caspase-9表达的影响。方法体外培养SH-SY5Y细胞,随机分为对照组、6-OHDA损伤组和SS-GAG（200、400和800mg/L）处理组,测定各组细胞中MDA含量、LDH活性,实时荧光（real-time）PCR检测细胞Bcl-2、Bax mRNA表达的变化,流式细胞技术（FCM）检测活化Caspase-9蛋白表达。结果与对照组相比较,6-OHDA损伤组的MDA含量、LDH活性增加,Bcl-2mRNA表达降低,Bax mRNA表达升高,表达活化Caspase-9蛋白的细胞数增加,差异均有统计学意义（F=27.699-280.567,P〈0.01）。与6-OHDA损伤组比较,各浓度SS-GAG处理组SH-SY5Y细胞中MDA含量、LDH活性降低,Bcl-2mRNA表达升高,Bax mRNA表达降低,活化的Caspase-9蛋白表达量减少,并且呈剂量依赖性,差异均有显著意义（F=27.699-280.567,P〈0.01）。结论 SS-GAG能降低6-OHDA诱导的氧化应激水平,保护生物膜的完整性。SS-GAG通过上调Bcl-2基因的表达和下调Bax基因的表达,抑制Caspase-9蛋白的活化,从而抑制6-OHDA诱导的SH-SY5Y细胞凋亡。Objective To investigate the effect of scallop skirt-glycosaminoglycan on the content of MDA and LDH and the expressions of several apoptosis-related genes in SH-SY5 Ycells induced by 6-OHDA. Methods Human neuroblastoma SHSY5 Ycells were cultured in vitro and divided into control group,6-OHDA-injured group and three groups treated with SS-GAG of200,400 and 800mg/L,respectively.The content of MDA and LDH were measured,the mRNA expressions of Bcl-2and Bax genes were evaluated by real-time PCR and the activation of Caspase-9was measured by flow cytometry. Results Compared with the control group,the items in the 6-OHDA destroy group were noted as follows：the content of MDA and LDH increased,the mRNA expression of Bcl-2reduced,the mRNA expression of Bax elevated and the number of cells expressing activated Caspase-9increased（F=27.699-280.567,P〈0.01）.Compared with the 6-OHDA-injured group,the content of MDA and LDH in SH-SY5 Ycells of each SS-GAG treated group decreased,the expression of Bcl-2mRNA of increased,the Bax mRNA expression declined,and the expression of activated Caspase-9reduced（F=27.699-280.567,P〈0.01）. Conclusion SS-GAG can reduce the damage of oxidative stress induced by 6-OHDA and protect the integrity of biofilms.SS-GAG inhibit the activation of Caspase-9by up-regulating Bcl-2and down-regulating Bax expression and thus inhibit the apoptosis of activation of SH-SY5 Ycells induced by 6-OHDA.

关 键 词：扇贝裙边糖胺聚糖 SH-SY5Y 氧化性应激 基因 BCL-2 基因 Bax CASPASE-9 

分 类 号：R741.02[医药卫生—神经病学与精神病学]