补肾安胎冲剂含药血清对大鼠骨髓来源内皮祖细胞增殖、迁移能力的影响  

Effect of serum containing Bushen Antai Granules( 补肾安胎冲剂) on proliferation andmigration ability of rat bone marrow derived endothelial progenitor cells

作　　者：杨春荣 余欣慧[2] 程红[2] 储继军[2] 李元琪 YANG Chunrong;YU Xinhui;CHENG Hong;CHU Jijun;LI Yuanqi(Gynecology Department,Community Health Service Center,Yuhua Economic Development Zone,Nanjing,Jiangsu,210012,China;Department of Gynecology,the First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei,Anhui,230012,China)

机构地区：[1]南京市雨花经济开发区社区卫生服务中心妇科,南京210012 [2]安徽中医药大学第一附属医院妇科,安徽合肥230012

出　　处：《甘肃中医药大学学报》2024年第2期1-7,共7页Journal of Gansu University of Chinese Medicine

基　　金：国家自然科学基金青年项目(81804133);安徽省高等学校科学研究项目(2023AH050762)。

摘　　要：目的观察不同浓度补肾安胎冲剂含药血清对大鼠骨髓来源内皮祖细胞(EPCs)增殖及迁移能力的影响。方法选取SPF级3~4周龄雌性SD大鼠,采用密度梯度离心法分离单个骨髓核细胞,收集培养7 d的细胞,分别通过双荧光染色法、免疫荧光法及流式细胞仪动态检测EPCs表面标志物等方法,鉴定分离培养的细胞为大鼠骨髓来源EPCs。制备补肾安胎冲剂含药血清,采用噻唑蓝(MTT)比色法检测不同浓度(5%,10%,15%,20%)补肾安胎冲剂含药血清作用不同时间(24,48,72 h)的大鼠骨髓来源EPCs的增殖率;通过Transwell法检测大鼠骨髓来源EPCs的迁移能力。结果经双荧光染色法鉴定,所培养细胞乙酰化低密度脂蛋白(Dil-AC-LDL)和异硫氰酸荧光素荆豆凝集素-1(FITC-UEA-1)双荧光染色呈双阳性;免疫荧光染色后,EPCs特异性表面抗原人白细胞分化抗原CD34、CD133及血管内皮生长因子受体2(VEGFR-2)染色均呈阳性,其阳性细胞率分别为94.7%,93.8%,98.8%,提示培养的细胞为EPCs。MTT比色法检测结果显示,经不同浓度的补肾安胎冲剂含药血清干预培养后,5%,10%,15%浓度含药血清可促进大鼠骨髓源性EPCs增殖,其中10%浓度含药血清作用48 h的细胞增殖率最高(50.34%);Transwell法检测结果显示,每高倍镜视野下对照组及含药血清组(10%)EPCs平均迁移细胞数分别为(51.0±7.94),(105.5±15.22)个,与对照组比较,含药血清组(10%)EPCs的迁移能力明显增强(P<0.01)。结论补肾安胎冲剂含药血清可增强大鼠骨髓源性EPCs的增殖及迁移能力,且10%浓度作用48 h EPCs的增殖率最高。Objective To observe the effects of different concentrations of serum containing Bushen Antai Granules(补肾安胎冲剂)on the proliferation and migration ability of rat bone marrow derived endothelial progenitor cells(EPCs).Methods The single bone marrow nuclear cells were isolated from SPF 3 to 4 weeks old female SD rats by density gradient centrifugation method,and the cells cultured for 7 days were collected.The isolated and cultured cells were identified as rat bone marrow derived EPCs by double fluorescence staining,immunofluorescence,a nd dynamic detection of EPCs surface markers by flow cytometry.The drug-containing serum of Bushen Antai Granules was prepared,and under the intervention of different concentrations(5%,10%,15%,20%)of serum containing Bushen Antai Granules at different times(24,48,72 h),the proliferation rate of bone marrow derived EPCs in rats was detected by MTT colorimetric method.The migration ability of bone marrow derived EPCs in rats was detected by Transwell method.Results The cultured cells were positive for Dil labeled acetylated low-density lipoprotein(Dil-AC-LDL)and fluorescein isothiocyanate-ulex europaeus lectin-1(FITC-UEA-1)by double fluorescence staining.After immunofluorescence staining,the EPCs specific surface antigens(human leukocyte differentiation antigen CD34,CD133 and VEGFR-2)were all positive,and the positive cell rates were 94.7%,93.8%and 98.8%,respectively,suggesting that the cultured cells were EPCs.The results of MTT coloration showed that 5%,10%and 15%concentrations of serum containing Bushen Antai Granules could promote the proliferation of bone marrow derived EPCs in rats,and the cell proliferation rate of 10%drug-containing serum at 48 h was the highest(50.34%).The results of Transwell method showed that the average number of EPCs migrating cells per high-power field of view was 51.0±7.94 in control group and 105.5±15.22 in drug-containing serum group(10%),respectively.Compared with the control group,the migration ability of EPCs in drug-containing serum g

关 键 词：补肾安胎冲剂 含药血清 内皮祖细胞 增殖能力 迁移能力 大鼠 实验研究 

分 类 号：R285.5[医药卫生—中药学]