欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
目的：检测糖尿病足伤口皮肤细胞凋亡情况，探讨晚期糖基化终产物（AGEs）对人皮肤成纤维细胞凋亡的影响。方法：选择36例足部伤口患者，糖尿病组18例，非糖尿病组18例。对2组临床和生化数据进行统计分析。采用免疫组化和TUNEL法检测伤口皮肤细胞凋亡情况。体外培养人原代皮肤成纤维细胞，分别给予正常浓度糖、持续高糖、波动高糖和AGEs干预72 h，采用蛋白质印迹法和流式细胞术测定细胞凋亡情况。结果：与非糖尿病组相比，糖尿病组血糖水平明显增高，伤口持续时间长（P＜0．05）。 Cleaved caspase-3在非糖尿病组和糖尿病组的免疫反应评分分别为1．04＆#177;0．23和3．04＆#177;0．31（P＜0．05）；TUNEL检测非糖尿病组和糖尿病组的细胞凋亡指数分别为（3．8＆#177;0．8）％和（8．4＆#177;1．5）％（ P＜0．05）。人原代皮肤成纤维细胞在正常浓度糖、持续高糖、高糖波动、AGEs干预下，cleaved caspase-3蛋白表达水平分别为0．80＆#177;0．13、1．22＆#177;0．18、1．46＆#177;0．32和1．83＆#177;0.25，凋亡率分别为（2．43＆#177;0．19）％、（2．89＆#177;0．51）％、（3．99＆#177;0．24）％和（6．83＆#177;0．36）％。 AGEs 组 cleaved caspase-3蛋白表达水平和凋亡率均明显高于正常浓度糖组和持续高糖组（P＜0．05）。结论：糖尿病皮肤创面细胞凋亡增加，可能是糖尿病皮肤伤口难愈的重要原因之一。与持续高糖、高糖波动组相比，AGEs促进人皮肤成纤维细胞凋亡的作用更强。
英文摘要：
AIM：Toinvestigatecellapoptosisindiabeticfootulcersandtheeffectofadvancedglycosylation end products （AGEs） on apoptosis in human fibroblast cells.METHODS： Diabetic foot patients （n=18） and 18 age-matched non-diabetic controls were recruited .The clinical and biochemical features were compared by statistics methods . Skin biopsies were obtained from foot .Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex （ SABC ） staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （ TUNEL） technique was used to detect apoptosis of the skin tissues .Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose （ changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h） or AGEs （150 mg/L, containing 5.6 mmo/L glucose）.After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3.Other cells were trypsinized , washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis .RE-SULTS：Diabetic patients had higher levels of fasting blood glucose （FBG）, 2-hour postprandial blood glucose （2 h PBG） and glycosylated hemoglobin A1c （HbA1c）, and longer wound duration.The protein level of cleaved caspase-3 was signifi-cantly higher in diabetic group , suggesting that apoptosis was increased in diabetic skin tissues .TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group （8.4%＆#177;1.5% vs 3.8%＆#177; 0.8%） , which further confirmed that cell apoptosis was increased in diabetic foot tissues .In human fibroblasts , the levels of cleaved caspase-3 in normal group , sustained high glucose group , fluctuant high glucose group and AGEs group were＆amp;nbsp;0.80 ＆#177;0.13, 1.22 ＆#177;0.18, 1.46 ＆#177;0.32 and 1.83 ＆#177;0.25, respectively.The apoptotic rates detected by flow
同期刊论文项目
同项目期刊论文
期刊信息
位置：