晚期糖基化终末产物与其受体对糖尿病创面氧化应激反应的影响  

Effects of advanced glycation end products and its receptor on oxidative stress in diabetic wounds

作　　者：牛轶雯[1] 缪明远[1] 董炜[1] 董叫云[1] 曹晓赞[1] 陆树良[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院上海市烧伤研究所,上海市刨面修复研究中心,200025

出　　处：《中华烧伤杂志》2012年第1期32-35,共4页Chinese Journal of Burns

基　　金：国家自然科学基金（30600645、30700871、81071568）

摘　　要：目的 观察糖尿病患者皮肤和创面中晚期糖基化终末产物（AGE）的蓄积和炎症反应,并结合体外干预实验推测两者之间可能存在的关系. 方法 采集10例2型糖尿病患者（糖尿病组）和12例非糖尿病患者（非糖对照组）的皮肤和创面组织标本,部分于光学显微镜下观察胶原排列、细胞分布（HE染色）,AGE及其受体（RAGE）表达（免疫组织化学染色）；部分匀浆后采用ELISA法检测丙二醛含量.体外采集、分离健康人外周血中性粒细胞,并分为正常对照组（添加RPMI-1640培养液培养）、低浓度干预组、中浓度干预组和高浓度干预组,3个干预组分别在RPMI-1640培养液中添加AGE修饰的人血清白蛋白,其中AGE的浓度依次为0.315、0.625、1.250 mg/mL;噻唑蓝法检测细胞存活率,荧光探针二氢二氯荧光黄双乙酸钠测定细胞内活性氧水平.对数据进行t检验. 结果 与非糖对照组比较,糖尿病组皮肤组织中胶原萎缩,排列紊乱；创面组织中炎性细胞弥散分布,胶原排列稀疏紊乱.糖尿病组皮肤组织中AGE和RAGE表达均多于非糖对照组；2组创面组织中RAGE呈阳性表达的细胞数量均多于所在组皮肤组织,以糖尿病组更显著,其创面内可见大量RAGE呈阳性表达的炎性细胞.糖尿病组患者皮肤和创面组织中每毫克蛋白丙二醛含量分别为（6 3±1.0）、（7.1±2.4）nmol,均明显高于非糖对照组相应组织[（2.9±1.0）、（3.6±1.4）nmol,t值分别为8.017、4.349,P ＜0.05或P＜0.01].与正常对照组[（34±5）％]比较,低浓度干预组、中浓度干预组、高浓度干预组中性粒细胞存活率均明显上升[（59±8）％、（77±5）％、（ 67±6）％,t值分别为7.195、14.890、11.130,P＜0.05或P＜0.01].与正常对照组（0.58±0.06）比较,低浓度干预组、中浓度干预组、高浓度干预组中性粒细胞内活性氧水平均升高（1.67±0.14、2.13±0.17、3 48±0.48,t值分别为20.195、24.905、16.8Objective To investigate the accumulation of advanced glycation end products（AGE）and the inflammatory response of skin and wound in diabetic patients,and to analyze their relationship in vitro. Methods Histological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type Ⅱ diabetes mellitus（diabetes group）and 12 non-diabetic patients with skin injury（control group）to observe the arrangement of collagen and the distribution of inflammatory cells,and to determine the expression levels of AGE and its receptor（ RAGE）.Malondialdehyde（MDA）levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay.In vitro,human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315,0.625,1.250 mg/mL,and they were identified as normal control（NC）group,low concentration（L）group,moderate concentration （M）group,and high concentration（H）group.Cell viability in each group was determined by MTT colorimetric assay,and the reactive oxygen species（ROS）in cell was measured with 2′,7′-dichlorfluorescein-diacetate.Data were processed with t test. Results Compared with those of skin in control group,collagens of skin tissues in diabetes group atrophied and disorderly arranged.Inflammatory cells in wounds in diabetes group were dispersed,in which collagens arranged loosely and irregularly,as compared with those of wounds in control group.Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group.In diabetes and control groups,especially in diabetes group,the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue.Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group.MDA level of skin and wound tissue in diabetes group was respectively（6.3 ± 1.0）,（7.1 ± 2.4）nmol per milligram protein

关 键 词：糖尿病 氧化性应激 糖基化终产物 高级 皮肤 创面愈合 

分 类 号：R587.2[医药卫生—内分泌]