皂角刺抑制人脐静脉血管内皮细胞增殖的实验研究

Experimental study on inhibition of proliferation of human umbilical vein endothelial cells by Spina Gleditsiae

目的研究皂角刺对离体的人脐静脉血管内皮细胞(HUVEC)增殖的影响,并从增殖细胞核抗原(PCNA)方面探讨其对HUVEC增殖抑制的原理。方法选择HUVEC为实验对象,取生长良好的HUVEC用于实验,利用100μmol/L二氯化钴(CoCl_2)诱导HUVEC增殖,同时分别将低、中、高的皂角刺作用于增殖状态下的HUVEC 24 h,进行以下的研究:(1)CCK-8法检测不同浓度皂角刺对HUVEC增殖的抑制程度;(2)流式细胞仪检测皂角刺对HUVEC核内PCNA表达的影响。结果 (1)空白组、增殖组、皂角刺低浓度组、皂角刺中浓度组、皂角刺高浓度组的OD值分别是1.542±0.048、1.836±0.053、1.463±0.042、1.160±0.052、0.861±0.047,示皂角刺显著抑制CoCl_2诱导的HUVEC增殖,而且有浓度依赖关系(P<0.01)。(2)空白组、增殖组、皂角刺低浓度组、皂角刺中浓度组、皂角刺高浓度组的PCNA表达率分别是55.29±2.44、79.95±1.62、49.90±4.62、29.15±3.42、18.15±2.53,示皂角刺可以明显降低HUVEC核内PCNA表达,从而抑制细胞增殖,并呈浓度依赖关系(P<0.01)。结论皂角刺有显著抑制HUVEC增殖的作用。

Objective To investigate the proliferative inhibition of human umbilical vein endothelial cells（ HUVEC） by Spina Gleditsiae and to further study its mechanism of the proliferative inhibition of HUVEC from Proliferating cell nuclear antigen（ PCNA）. Methods HUVEC were cultured and were used for experiment in good condition. HUVEC proliferation were induced by 100 μmol/L CoCl2,in the same time 20 mg/ml,40 mg/ml,80 mg/ml Spina Gleditsiae were added to the HUVEC separately. The following studies were made after 24 hours incubation： 1. The effects of proliferative inhibiton by Spina Gleditsiae on the HUVEC were assayed with CCK-8 method. 2. The changes of PCNA expression of HUVEC regulated by Spina Gleditsiae were assayed with flow cytometry. Results 1. In the CCK-8 inspection experiment,the OD values of blank control group,proliferation control group,low concentration group,middle concentration group,high concentration group are 1. 542 ± 0. 048,1. 836 ± 0. 053,1. 463 ± 0. 042,1. 160 ± 0. 052,0. 861 ± 0. 047 respectively; Spina Gleditsiae can significantly inhibit the proliferation of HUVEC induced by CoCl2 and these effects were dose dependent（ P〈0. 01）. 2. In the flow cytometry inspection experiment,the PCNA expression rates of blank control group,proliferation control group,low concentration group,middle concentration group,high concentration group are 55. 29± 2. 44,79. 95 ± 1. 62,49. 90 ± 4. 62,29. 15 ± 3. 42,18. 15 ± 2. 53 respectively; Spina Gleditsiae can downregulate the expression of PCNA in HUVEC and this decrease was dose dependent（ P〈0. 01）. Conclusion Spina Gleditsiae has significantly an inhibitive effect on the HUVEC proliferation.