摘要
目的观察自噬抑制剂3-甲基腺嘌呤(3-MA)调控大鼠骨髓源性内皮祖细胞(EPCs)自噬,探讨其对静脉血栓再通的影响。方法密度梯度法取大鼠骨髓EPCs,体外诱导分化并鉴定。建立大鼠静脉血栓模型。实验分为4组:单纯生理盐水组(对照组,A组)、单纯3-MA组(5mmol/L3-MA,B组)、单纯EPCs组(C组)和3-MA联合EPCs组(5mmol/L3-MA作用于EPCs24h,D组),每组各20只大鼠。在血栓形成后的10天经大鼠尾静脉注入各组试剂,再过14天取标本。苏木精-伊红染色(HE)及vWF免疫组化染色观察血栓机化再通情况,并在镜下计数各组毛细血管数目,取其平均值。结果 (1)D组经HE及免疫组化染色观察发现新生血管最多,与其余各组比较差异均有统计学意义(P<0.05);(2)C组分别与A组和B组比较,差异有统计学意义(P<0.05);(3)A组与B组比较,差异无统计学意义(P>0.05)。结论 5mmol/L3-MA调控自噬后的EPCs经大鼠尾静脉注射可促进大鼠静脉血栓再通,且比单纯注射EPCs效果更好。
Objective To investigate the effect of 3-MA on promoting recanalization of venous thrombosis by regulating autophagy of rat endothelial progenitor cells(EPCs). Methods Bone marrow-derived mononuclear cells were isolated from rat bone marrow by Ficoll. Rat model of venous thrombosis was established (80 rats).There were four groups, including the nature saline group(group A, NS, n =20), the simple 3-MA group (group B, 5 mmol/L 3-MA, n =20), the simple EPCs group(group C, n =20),and the 3-MA together with EPCs group(group D, 5 mmol/L 3-MA and EPCs, n =20).Each group reagents were injected through rat tail 10 days after thrombosis formation and specimens were taken 14 days later. HE and immunohistochemical staining were used to observe recanalization of thrombosis,and the number of capillaries in each group were counted and averaged. Results Neovascularization was most in group D by HE and immunohistochemical staining observation compared with other groups (P < 0.05).Neovascularization of group C compared to group B and group A, respectively, and the differences were statistically significant (P < 0.05). Neovascularization of group A compared to group B, the differences were not statistically significant (P > 0.05). Conclusion 5 mmol/L 3-MA together with EPCs injection through rat tail vein promotes recanalization of venous thrombosis by regulating autophagy, and its effect is better than simple EPCs injection.
出处
《中国血管外科杂志（电子版）》
2013年第1期47-50,共4页
Chinese Journal of Vascular Surgery(Electronic Version)
基金
国家自然科学基金资助项目(30972941)
关键词
内皮祖细胞
3-甲基腺嘌呤
静脉血栓
自噬
Endothelial progenitor cells
3-methlyadenine
Venous thrombosis
Autophagy
