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目的探讨内皮祖细胞(EPC)与雌激素联合应用防止PCI术后再狭窄的效果。方法雌性新西兰大白兔60只随机分为5组,每组12只。Ⅰ:假手术组;Ⅱ:对照组;Ⅲ:雌激素组;Ⅳ:EPC组;Ⅴ:雌激素+EPC组。卵巢切除术后,Ⅲ组和Ⅴ组给予雌激素替代治疗,其余动物皮下注射生理盐水,卵巢切除前及卵巢切除后3d、2周分别采血,应用放免法检测血清雌激素水平。实验动物高脂饮食6周后应用氧化酶法检测血浆总胆固醇、甘油三酯水平;用直接法检测低密度脂蛋白胆固醇及高密度脂蛋白胆固醇。从兔外周血分离培养单个核细胞,制备EPC悬液以备细胞移植。建立兔腹主动脉损伤模型,Ⅰ组动物仅分离暴露出股动脉而不做球囊损伤,Ⅳ组和Ⅴ组动物损伤血管局部经导管给予EPC悬液2mL输注,其余动物(除Ⅰ组外)给予生理盐水2mL,4周后,伊文斯蓝染色检测损伤血管段内皮修复程度,免疫组化法检测细胞增殖程度,HE染色观察管腔丢失情况。结果与Ⅰ、Ⅱ、Ⅳ组相比,Ⅲ、Ⅴ组雌激素水平明显升高(P<0.01),总胆固醇、低密度脂蛋白胆固醇明显降低(P<0.01),高密度脂蛋白胆固醇明显升高(P<0.15),甘油三脂差异无统计学意义(P>0.05)。Ⅴ组与Ⅲ、Ⅳ组相比,内皮修复程度、细胞增殖程度及管腔丢失程度差异有统计学意义,分别为[(90.2±5.1)%vs(85.1±4.1)%、(83.9±6.5)%,P<0.05],[(15.67±2.98)%vs(26.02±4.18)%、(24.87±3.54)%,P<0.01],[(17.61±2.69)%vs(38.43±4.35)%、(41.15±4.45)%,P<0.01]。结论EPC与雌激素联合应用能加速内皮修复,减少血管腔丢失。
Objective To investigate the effect of endothelial progenitor cells (EPCs) in combination with estrogen to prevent restenosis after PCI. Methods Sixty female New Zealand white rabbits were randomly divided into five groups (n = 12). Ⅰ: sham operation group Ⅱ: control group Ⅲ: estrogen group Ⅳ: EPC group V: estrogen + EPC group After ovariectomy, patients in groups Ⅲ and Ⅴ were treated with estrogen replacement therapy. The other animals were injected subcutaneously with normal saline, before ovariectomy and at 3d and 2 weeks after ovariectomy. Serum estrogen levels were determined by radioimmunoassay. Six weeks after the experiment of high-fat diet, the levels of total cholesterol and triglyceride in plasma were detected by oxidase method. The levels of LDL-C and HDL-C were measured by direct method. Mononuclear cells were isolated and cultured from peripheral blood of rabbits to prepare EPC suspension for cell transplantation. Rabbits in abdominal aorta injury model were established. Group Ⅰ animals only exposed femoral artery without balloon injury. Animals in group Ⅳ and groupⅤwere given 2 mL infusion of EPC suspension via local catheterization. The animals except groupⅠ ) Were given 2mL of saline. After 4 weeks, Evans blue staining was used to detect the degree of endothelium repair in injured vascular segments. The degree of cell proliferation was detected by immunohistochemistry and the lumen loss was observed by HE staining. Results Compared with Ⅰ, Ⅱ and Ⅳ groups, the levels of estrogen in groups Ⅲ and Ⅴ increased significantly (P <0.01), total cholesterol and low density lipoprotein cholesterol decreased significantly (P <0.01), and high density lipoprotein cholesterol increased significantly High (P <0.15), triglyceride difference was not statistically significant (P> 0.05). Compared with group Ⅲ and group Ⅳ, the degree of endothelium repair, the degree of cell proliferation and the degree of lumen loss in group V were statistically significant, which were (90.2 ± 5.1)% vs (85.1 ± 4.1)%, (83.9 ± 6.5) %, P <0.05], [(15.67 ± 2.98)% vs (26.02 ± 4.18)%, (24.87 ± 3.54)%, P <0.01], [(17.61 ± 2.69)% vs (38.43 ± 4.35)%, 41.15 ± 4.45)%, P <0.01]. Conclusions The combination of EPC and estrogen can accelerate endothelial repair and reduce the loss of blood vessel.