扇贝多肽对H_2O_2诱导HaCaT细胞氧化损伤的作用  

EFFECTS OF PCF ON H_2O_2-INDUCED HACAT CELL OXIDATIVE DAMAGE

作　　者：赵慧慧[1] 王道艳[1] 郝冉冉[1] 党转宁 韩彦弢[1] 

机构地区：[1]青岛大学医学院药学院,山东青岛266071

出　　处：《青岛大学医学院学报》2014年第4期289-291,295,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省自然科学基金资助项目(NO.ZR2011HM-046)

摘　　要：目的了解扇贝多肽对H2O2诱导的HaCaT细胞氧化损伤的保护作用及其机制。方法取对数生长期HaCaT细胞,随机分为正常对照组（细胞正常培养）、模型组（200μmol/L的H2O2作用12h）、扇贝多肽不同浓度组（低、中、高浓度,分别用1.42、2.84、5.68mmol/L的扇贝多肽预处理2h后,再给予200μmol/L的H2O2作用12h）,应用Hochest33258染色方法检测各组细胞凋亡率,CCK8检测细胞存活率,以二氢荧光黄双乙酸盐（DCFH-DA）为荧光探针、流式细胞术检测细胞内的活性氧（ROS）含量,蛋白印迹法检测生长阻滞和DNA损伤诱导修复基因45a（Gadd45a蛋白）的表达。结果与正常对照组比较,模型组的细胞凋亡率明显升高（F=439.80,q=38.24,P〈0.01）,细胞存活率下降（F=191.40,q=214.10,P〈0.01）,细胞内ROS含量增加（F=121.10,q=31.02,P〈0.01）,Gadd45a蛋白表达升高（F=66.59,q=14.57,P〈0.01）;与模型组比较,扇贝多肽各浓度组细胞凋亡率明显降低（q=9.60～25.82,P〈0.01）,细胞存活率升高（q=13.47～54.86,P〈0.01）,细胞内ROS减少（q=4.20～12.14,P〈0.01）,Gadd45a蛋白表达降低（q=4.04～15.55,P〈0.01）;扇贝多肽各浓度组间比较,随扇贝多肽浓度升高,细胞凋亡率下降,细胞存活率升高,细胞内ROS含量降低,Gadd45a的表达降低,差异有显著性（q=3.96～23.36,P〈0.05）。结论 1.42～5.68mmol/L扇贝多肽能够清除细胞内的ROS,抑制Gadd45a蛋白的表达,从而保护H2O2诱导的HaCaT细胞氧化应激损伤。Objective To investigate the protection of PCF on H2O2-induced oxidative stress damage of HaCaT cells and its mechanism. Methods Log-phase-growth cells were collected and randomized to normal control group（the cells were normally cultured）,model group（pretreated with 200μmol/L H2O212h）,low-,moderate-and high-concentration groups（pretreated with 1.42mmol/L PCF,2.84mmol/L PCF and 5.68mmol/L PCF for 2h,respectively,and then treated with 200μmol/L of H2O2for 12h）.The Hoechst33258staining,CCK8method,flow cytometry and western blot were used to investigate the apoptosis condition,cell viability,generation of reactive oxygen intermediates and expression of Gadd45a,respectively. Results Compared with the control group,the rate of apoptotic in the model group significantly increased（F=439.8,q=38.24,P〈0.01）,the cell survival decreased（F=191.40,q=214.10,P〈0.01）,intra-cellular ROS content increased（F=121.10,q=31.02,P〈0.01）and Gadd45aprotein expression elevated（F=66.59,q=14.57,P〈0.01）.Compared with the model group,the apoptosis rate in each PCF group was much lower（q=9.60-25.82,P〈0.01）,cell survival increased（q=13.47-54.86,P〈0.01）,intra-cellular ROS decreased（q=4.20-12.14,P〈0.01）and Gadd45aprotein expression elevated（q=4.04-15.55,P〈0.01）.A comparison between each-different-concentration PCF groups showed that along with the increase of PCF concentration,the apoptosis declined,cell survival increased,intracellular ROS lowered and Gadd45aprotein expression reduced,the differences were significant（q=3.96-23.36,P〈0.05）. Conclusion PCF（1.42-5.68mmol/L）can clear intra-cellular ROS and inhibit Gadd45aexpression and thus protect HaCaT cells from the oxidative stress injury induced by H2O2.

关 键 词：扇贝多肽 细胞凋亡 活性氧 生长阻滞和DNA损伤诱导修复基因45a HACAT细胞 

分 类 号：R285[医药卫生—中药学]