脐带间充质干细胞上清提取物对糖尿病皮肤溃疡大鼠肿瘤坏死因子-α及血管内皮生长因子的影响  

Effect of supernatant of umbilical cord mesenchymal stem cells on tumor necrosis factor-α and vascular endothelial growth factor in diabetic skin ulcer rats

作　　者：徐海环[1,2] 董化江[1] 赵明亮 XU Hai-huan DONG Hua-jiang ZHAO Ming-liang(State Key Laboratory of Precision Measurement Technology and Instruments, Tianjin University, Tianjin 300072, China Department of Dermatology, the Affiliated Hospital of Logistics University of Chinese People＇s Armed Police Forces, Tianjin 300162, China Neural Restoration Key Laboratory of Tianjin, Tianjin 300162, China)

机构地区：[1]天津大学精密测试技术及仪器国家重点实验室,天津300072 [2]武警后勤学院附属医院皮肤科,天津300162 [3]天津市神经修复重点实验室,天津300162

出　　处：《新乡医学院学报》2017年第2期90-93,共4页Journal of Xinxiang Medical University

基　　金：天津市自然科学基金资助项目(编号:16JCYBJC27600);武警后勤学院附属医院种子基金项目(编号:FYM201519)

摘　　要：目的探讨脐带间充质干细胞上清提取物对糖尿病皮肤溃疡大鼠肿瘤坏死因子-α(TNF-α)及血管内皮生长因子(VEGF)的影响。方法将30只Sprague-Dawley大鼠随机分为模型组、单纯培养基冻干粉治疗组和间充质干细胞上清冻干粉治疗组,每组10只。大鼠经尾静脉注射四氧嘧啶并给予高脂饲料喂养制作糖尿病模型。糖尿病大鼠采用戊巴比妥钠麻醉后,用刀片在小腿处划一切口,深达皮下,注入金黄色葡萄球菌悬液,造成皮肤溃疡。皮肤溃疡模型制作成功后模型组大鼠尾静脉注射100μL生理盐水,溃疡处滴加100μL生理盐水;单纯培养基冻干粉治疗组大鼠将培养基冻干粉溶解至200μL生理盐水,尾静脉注射100μL,溃疡处滴加100μL;间充质干细胞上清冻干粉治疗组大鼠将间充质干细胞上清冻干粉溶解至200μL生理盐水,尾静脉注射100μL,溃疡处滴加100μL。5 d后采用放射免疫法检测大鼠血清TNF-α水平,采用反转录-聚合酶链式反应和Western blot检测大鼠溃疡处组织中TNF-α和VEGF的表达。结果间充质干细胞上清冻干粉治疗组大鼠血清TNF-α水平显著低于模型组和单纯培养基冻干粉治疗组(P<0.05);模型组与单纯培养基冻干粉治疗组大鼠血清TNF-α水平比较差异无统计学意义(P>0.05)。间充质干细胞上清冻干粉治疗组大鼠溃疡组织中TNF-αmRNA及蛋白表达低于模型组和单纯培养基冻干粉治疗组(P<0.05);间充质干细胞上清冻干粉治疗组大鼠溃疡组织中VEGF mRNA及蛋白的表达高于模型组和单纯培养基冻干粉治疗组(P<0.05);模型组大鼠溃疡组织中TNF-αmRNA、VEGF mRNA及相应蛋白表达与单纯培养基冻干粉治疗组比较差异无统计学意义(P>0.05)。结论间充质干细胞上清冻干粉治疗可有效降低溃疡组织中TNF-α的表达,且可有效提高溃疡组织中VEGF的表达水平。Objective To study the effect of supernatant of umbilical cord mesenchymal stem cells（ MSCs） on tumor necrosis factor-α（ TNF-α） and vascular endothelial growth factor（ VEGF） in diabetic skin ulcer（ DSU） rats. Methods Thirty Sprague-Dawley rats were randomly divided into model group,culture medium treatment group and MSCs supernatant treatment group,with 10 rats in each group. The diabetic rat models were made by injecting alloxan through caudal vein and giving highfat diet. The diabetic rats were anesthetized with pentobarbital sodium; and then were cut a wound by the blade in the calf,the depth to the subcutaneous tissue. The wounds were infused the suspension of Staphylococcus aureus to form the skin ulcer. After eatblishing the skin ulcer models,the rats in model group were injected physiological saline 100 μL through caudal vein; at the same time,the physiological saline 100 μL was droped to the ulcer tissure. In the culture medium treatment group,the freezedried powder of culture medium was dissolved in 200 μL physiological saline,an then 100 μL was infused to rats through caudal vein,100 μL was droped to the ulcer tissure. In MSCs supernatant treatment group,the freeze-dried powder of supernatant was dissolved in 200 μL physiological saline,and then 100 μL was infused to rats through caudal vein,100 μL was droped to the ulcer tissure. After five days,the serum levels of TNF-α was detected by radioimmunoassay; the expression of VEGF and TNF-α in ulcer tissure of rats was measured by reverse transcription polymerase chain reaction and Western blotting. Results The serum level of TNF-α of rats in MSCs supernatant treatment group was significanlty lower than that in model group and culture medium treatment group（ P 0. 05）; there was no statistic difference in serum level of TNF-α of rats between the model group and culture medium treatment group（ P 0. 05）. The expressions of TNF-α mRNA and protein in ulcer tissure of rats in MSCs supernatant treatment group were significan

关 键 词：脐带 间充质干细胞 糖尿病皮肤溃疡 肿瘤坏死因子-α 血管内皮生长因子 大鼠 

分 类 号：R587.1[医药卫生—内分泌] R753[医药卫生—内科学]