晚期糖基化终产物对脂肪干细胞功能、血管新生能力的影响及丹红注射液保护作用的实验研究  

Effect of Advanced Glycation End Products on the Function and Angiogenesis of Adipose Tissue-Derived Stem Cells and the Protective Effect of Danhong Injection:an Experimental Study

作　　者：伍锋[1] 贺治青[1] 纪睿圳 王新[1] 梁春[1] 吴宗贵[1] 

机构地区：[1]中国人民解放军第二军医大学附属长征医院心血管内科,上海200003

出　　处：《中国中西医结合杂志》2014年第7期839-845,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81130065);国家自然科学基金面上项目(No.81270405)

摘　　要：目的观察糖尿病机体主要的晚期糖基化终产物亚型-Nε羧甲基赖氨酸修饰白蛋白[Nε-(carboxymethyl)lysine albumin,CMLs]对脂肪干细胞(adipose tissue-derived stem cells,ADSCs)功能及血管新生作用的影响及丹红注射液(Danhong Injection,DH)的保护作用。方法采用酶消法从人皮下脂肪组织中分离培养人ADSCs,并通过光学显微镜进行形态学观察、分化能力评估。进一步采用WST-1增殖试剂盒、Transwell小室、流式AnnexinⅤ-FITC/PI凋亡检测试剂盒以及人VEGF的ELISA试剂盒分别观察对照组、BSA组(60μg/mL)、CML-BSA组(60μg/mL)、DH组(100μL/mL)、联合用药组(60μg/mL CML-BSA加100μL/mL DH),共5组干预液对ADSCs增殖、迁移、凋亡和分泌功能的影响。并通过体外血管新生实验观察其对ADSCs血管新生能力变化情况的影响。结果与BSA组比较,CML-BSA组能显著抑制ADSCs增殖和迁移的能力,增加凋亡,减少VEGF的分泌,减弱其血管新生能力(P<0.05);与对照组比比较,100μl/mL DH能显著促进ADSCs增殖和迁移的能力,抑制其凋亡,增加VEGF的分泌,改善其血管新生能力(P<0.05);与CML-BSA组比较,联合治疗组能显著逆转CML-BSA对ADSCs增殖和迁移能力的抑制作用,改善CML-BSA对ADSCs凋亡的促进作用,增加VEGF的分泌,提高其血管新生能力(P<0.05)。结论 CMLs能抑制ADSCs的增殖和迁移能力,促进其凋亡,抑制其血管新生能力。DH能改善CMLs的作用。Objective To investigate the effect of Nε-（carboxymethyl） lysine albumin （CMLs）, a primary advanced glycation end products （AGEPs） isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells （ADSCs） and the protective effect of Danhong Injection （DH） Methods Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 μg/mL BSA, 60 μg/mL CML-BSA, 100μL/mL DH, and 60 μg/mL CML-BSA ＋100 μL/rnL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test. Results Compared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened （P 〈0.05）. Compared with the blank control group, 100 μL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs （P 〈0.05）. Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated （P 〈0.05）. Conclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

关 键 词：丹红注射液 脂肪干细胞 晚期糖基化终产物 血管新生 

分 类 号：R587.1[医药卫生—内分泌]