应用高通量蛋白芯片技术筛查糖尿病小鼠创面差异蛋白的研究  

Study on Screening Differential Protein of Wound in Diabetic Mice with High Throughput Chip

作　　者：刘鹏[1,2] 徐阳 刘国涛[2] 周玉 王军 LIU Peng;XU Yang;LIU Guo-tao(Graduate School,Tianjin University of Traditional Chinese Medicine,Tianjin(301617),China)

机构地区：[1]天津中医药大学第一附属医院/国家中医针灸临床医学研究中心,天津300193 [2]天津中医药大学研究生院,天津301617

出　　处：《中国中西医结合外科杂志》2021年第6期819-825,共7页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金(81973854);天津市自然科学基金项目(18JCYBJC93600)。

摘　　要：目的:应用高通量蛋白芯片技术筛查糖尿病小鼠创面与正常小鼠创面组织的差异蛋白,探讨糖尿病性创面难愈的可能机制。方法:选取雄性健康BALB/c小鼠20只,随机选取10只予常规饲喂作为正常组,余10只为糖尿病组,高脂高糖联合注射链脲佐菌素(STZ)法制备糖尿病模型,皮肤全层切除法制备创面模型,在创面模型复制成功第3天取材。GSM-CAA-4000芯片定量检测两组小鼠创面组织中200种蛋白因子的含量。结果:高脂高糖饲喂联合腹腔注射STZ法成功制备糖尿病小鼠模型,糖尿病组小鼠成模后体重明显下降(P<0.05),血糖高于正常组(P<0.01)。GSM-CAA-4000芯片定量检测两组小鼠创面200种蛋白水平,共筛选出29种差异蛋白。与正常组对比,糖尿病组创面组织中含量降低的蛋白因子包括:ACE、Gas 1、IL-33、P-Cadherin、E-selectin、6Ckine、IL-1ra、Periostin、VEGF-D、Dtk、MCSF、SDF-1a、VEGF、TARC、Pro-MMP-9、gp130、VCAM-1、TNF RI(P<0.05);含量升高的蛋白因子包括:MBL-2、CD36、SHH-N、Renin 1、Fetuin A、Limitin、IL-10、VEGF-B、Persephin、TRANCE(P<0.05)。结论:通过蛋白芯片技术共筛选出29个差异蛋白,主要涉及生长因子、趋化因子、黏附分子、肿瘤坏死因子及受体家族,提示糖尿病性创面存在细胞增殖、趋化,迁移黏附以及基质降解等方面异常。Objective To investigate the possible mechanism of diabetic wound recovery by screening the differential protein between diabetic mice and normal mice with high-throughput protein microarray.Methods There were 20 healthy BALB/c male mice,10 of them were randomly selected as the normal group and the remaining 10 were randomly selected as the diabetic group.The diabetic model was prepared by STZ injection combined with high fat and high sugar,and the wound model was prepared by full-thickness skin resection.The samples were collected on the third day after the successful replication of the wound model.GSMCAA-4000 microarray was used to quantitatively detect the content of 200 protein factors in the wound tissues of two groups of mice.Results The diabetic mouse model was successfully prepared by high fat and high sugar feeding combined with intraperitoneal injection of STZ.The body weight of the diabetic wound group decreased significantly(P<0.05),and the blood glucose of the diabetic wound group was higher than that of the normal group(P<0.01).GSM-CAA-4000 microarray was used to quantitatively detect 200 proteins in the wound surface of mice of the two groups,and a total of 29 different proteins were screened out.Compared with the normal group,the protein factors that increased in the diabetic group included ACE,GAS 1,IL-33,P-cadherin,E-selectin,6 Ckine,IL-1 ra,Periostin,VEGF-D,Dtk,MCSF,SDF-1 a,VEGF,TARC,Pro-MMP-9,gp 130,VCAM-1,TNF RI(P<0.05).The reduced protein factors included MBL-2,CD 36,SHH-N,Renin 1,Fetuin A,Limitin,IL-10,VEGF-B,Persephin and TRANCE(P<0.05).Conclusion A total of 29 differential proteins were screened by protein chip technology,mainly involving growth factors,chemokines,adhesion molecules,tumor necrosis factor and receptor families,suggesting abnormal cell proliferation,chemotaxis,migration adhesion and matrix degradation in diabetic wounds.

关 键 词：高通量芯片 糖尿病 创面愈合 

分 类 号：R659[医药卫生—外科学]