摘要:
目的 观察通心络联合外周血干细胞移植治疗对糖尿病足溃疡大鼠促血管新生的影响.方法 2012年10月—2013年8月于河北以岭医药研究院药理实验室进行实验.高脂喂养联合低剂量链脲佐菌素(STZ)建立糖尿病大鼠模型,通过冰块降温和液氮冷冻,诱导大鼠糖尿病足溃疡模型.将造模成功的50只大鼠分为正常对照组、模型组、通心络组、干细胞组、通心络+干细胞组5组,每组10只,正常对照组和模型组大鼠以等体积的生理盐水灌胃;通心络组给予通心络0.8 g/kg灌胃,1次/d;干细胞组给予干细胞移植治疗;通心络+干细胞组给予干细胞移植同时给予通心络0.8 g/kg灌胃,1次/d,于给药7 d后处死动物取材进行指标检测.ELISA法测定血清血管内皮生长因子(VEGF)、晚期糖基化终末产物(AGE)、晚期糖基化终末产物受体(RAGE),RT-PCR法测定组织局部VEGF mRNA、RAGE mRNA表达水平.结果 各组间比较血清VEGF、AGE、RAGE有显著差异(F=28.675、18.018、35.946,P均=0.000),与模型组比较,通心络组、干细胞组及通心络+干细胞组血清VEGF显著增加(P0.05);与干细胞组比较,通心络组显著增加溃疡局部VEGF mRNA表达(P<0.01),降低RAGE mRNA表达(P<0.05),通心络+干细胞组显著增加溃疡局部VEGF mRNA表达(P=0.002),显著降低RAGE mRNA表达(P<0.01);与通心络组比较,通心络+干细胞组增加溃疡局部VEGF mRNA表达(P<0.05),降低RAGE mRNA表达(P<0.05).结论 通心络联合外周血干细胞移植能够提高糖尿病足溃疡大鼠血清和溃疡局部组织中VEGF水平,降低AGE、RAGE水平,促进血管新生.%Objective To discuss promoting angiogenesis by observing the levels of vascular endothelial growth factor (VEGF),advanced glycation end products(AGE),advanced glycation end products receptor(RAGE) in the diabetic foot ul-cers rats treated by Tongxinluo capsule combined with peripheral blood stem cell transplantation. Methods From October 2012 to August 2013,the research was conducted in Hebei Yiling Medicine Pharmacology Laboratory. The diabetic rat model was es-tablished. Diabetic foot ulcers rats were randomly divided into five groups(n=10):normal control,model,TXL,stem cell, TXL + stem cells,the serum level of VEGF,AGE,RAGE were determined by enzyme linked immunosorbent assay,the local organizations levels of VEGF mRNA,RAGE mRNA expression were examined by RT PCR method. Results The comparison between groups of serum VEGF,AGE and RAGE have significant difference (F=27.675,F=18.018,F=35.946,P=0. 000),compared with model group, Tongxinluo capsule group and Tongxinluo capsule + stem cell group serum VEGF in-creased significantly (P=0.003, P=0.000), AGE, RAGE decreased significantly (P=0.006, P=0.000), stem cell group increased(P=0.015),AGE and RAGE decreased(P=0.036,P=0.021);Tongxinluo capsule group showed signif-icantly increased serum VEGF (P=0.006), AGE and RAGE decreased (P=0.017, P=0.030), Tongxinluo capsule +stem cell group's serum VEGF increased significantly (P=0.000),AGE,RAGE significantly decreased (P=0.002, P=0.000);compared with Tongxinluo capsule group,Tongxinluo capsule + stem cell group showed increased of serum VEGF (P=0.041), AGE and RAGE decreased (P =0.035, P =0.013); Among the groups, ulcer tissue VEGF mRNA and RAGE mRNA expression have significant difference (F=8.152,F=7.650, P=0.000), compared with the model group, Tongxinluo capsule group increased the expression of VEGF mRNA and decreased the expression of RAGE mRNA (P=0. 014,P=0.023), Tongxinluo capsule + stem cells group significantly increased the expression of VEGF mRNA and de-creased the expression of RAGE mRNA (P=0.003, P=0.000); in stem cell group, the expression of RAGE mRNA and VEGF mRNA in ulcer tissue showed no significant difference compared with the model group(P=0.235,P=0.470);com-pared with the stem cell group,Tongxinluo capsule group significantly increased the expression of VEGF mRNA in local ulcer (P=0.008) and decreased the expression of RAGE mRNA (P=0.039), Tongxinluo capsule + stem cells group signifi-cantly with increased local VEGF mRNA expression(P=0.002),significantly reduced the expression of RAGE mRNA(P=0.001); compared with Tongxinluo capsule group, Tongxinluo capsule + stem cell group increased ulcer's expression of VEGF mRNA (P=0.032),reduced the expression of RAGE mRNA (P=0.027). Conclusion Tongxinluo capsule com-bined peripheral blood stem cell transplantation can improve the VEGF levels in serum and local organizations of diabetic foot ulcers rats,and reduce AGE,RAGE levels,promoting angiogenesis.