体外低能量震波对人脐静脉内皮细胞和血管内皮生长因子及白细胞介素-8变化的影响  

Effects of low-energy extracorporeal shock wave on human umbilical vein endothelial cells,vascular endothelial growth factor and interleukin-8 expression

作　　者：李丽[1] 马一铭 李琳[1] 胡钊[1] 郭涛[1] 肖践明[1] 蔡红雁[1] 

机构地区：[1]昆明医科大学第一附属医院心内科,昆明医学硕士研究生650032

出　　处：《医学研究生学报》2015年第8期789-793,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81260027);云南省卫生厅科技计划项目(2012WS005);云南省科技计划项目(2014FZ023)

摘　　要：目的体外震波治疗能促进缺血组织的血管新生和加速侧支循环建立,但其具体作用机制尚不明确。探讨0.09 m J/mm2的体外震波对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖及血管内皮生长因子(vascular endothelial growth factor,VEGF)、白细胞介素-8(interleukin-8,IL-8)变化的影响。方法实验分为实验组和对照组,对照组试管置于震波仪器中但不进行震波处理,实验组以0、0.03、0.09、0.18、0.24 m J/mm2震波能量冲击,每孔加CCK8溶液,用酶标仪测定在波长450 nm处的A值。对体外培养的HUVECs细胞株施予0.09 m J/mm2能量的震波处理,CCK8比色法检测细胞增殖情况,Real-time PCR法和流式细胞术分别检测低能量超声波处理后细胞因子VEGF、IL-8 mRNA表达和蛋白质的变化。结果 1CCK8比色法检测结果显示实验组中只有0.09 m J/mm2能量A450值与对照组比,差异有统计学意义(0.70±0.04 vs0.54±0.09,P<0.05),而其他能量与对照组比,差异无统计学意义(P>0.05);2RT-PCR结果显示0.09 m J/mm2能量时HUVECs VEGF、IL-8 mRNA的表达(7.93±0.90、7.34±1.67)均较对照组(1.07±0.40、1.00±0.09)明显升高(P<0.01)。3流式细胞术检测结果显示与对照组相比,0.09 m J/mm2能量时HUVECs VEGF、IL-8蛋白表达(39.89±4.79、31.33±5.61)均较对照组(20.98±3.30、22.60±3.76)明显升高(P<0.05)。结论体外低能量震波可促进HUVECs增殖和增加VEGF、IL-8的表达,VEGF、IL-8的表达增加可在体外震波促进血管新生机制中起重要作用。Objective Extracorporeal shock wave therapy （ESWT） can promote angiogenesis and collateral circulation es- tablishment by introducing shock waves to ischemia myocardium, however, the specific mechanisms remain unclearly. The study aimed to explore the effects of 0.09mJ/mm2 shock wave treatment on human umbilical vein endothelial cell （HUVEC） proliferation, vascular endothelial growth factor （VEGF） and Interleukin-8 （IL-8） expressions. Methods There were an experimental group and a control group in the experiment. The tubes of the experiment group were set in shock wave devices without the treatment of shock waves. As for the experiment group, 0, 0.03, 0.09, 0.18, 0.24 mJ/mm2 shock waves were introduced, followed by the addition of CCK8 solution and the measurement of A value at the wavelength of 450nm by microplate reader. The HUVEC cell lines were performed 0.09 mJ/mm2 ultrasonic shock energy, CCK8 colorimetric method were utilized to detect the HUVEC proliferation, real time PCR and flowcytometry were applied to detect the expressions of VEGF and IL-8 mRNA respectively. Results ① CCK8 colorimetric method revealed that 0.09 mJ/mm2 shock energy markedly promoted the HU- VEC proliferation compared with the control group, with A450 value comparison （0.70 ±0.04 vs 0. 54 ±0.09, P 〈 0.05 ）, while no sta- tistical significance was found in 0.03, 0. 18, 0.24 mJ/mm2 shock energy compared with the control group （ P 〉 0. 05 ）. ② RT-PCRshowed that the 0. 09 mJ/mm2 energy significantly enhanced the expressions of VEGF and IL-8 mRNA compared with the control group （9.93 ± 0. 90 vs 1.09± 0.40, 7.34± 1.67 vs 1. 00 ± 0.09, P 〈 0. 001 ）.③Flow Cytometry showed that the expressions of VEGF and IL-8 mRNA significantly increased after 0.09 mJ/mm2ESWT compared with the control group（39.89 ±4.79 vs 20.98±3.30, 31.33 ±5.61 vs 22.60± 3.76, P 〈 0.05）. Conclusion Low-energy ESWs can surely promote HUVEC proliferation and increase the expres- sions of VEGF and IL-8, and the up-regnlati

关 键 词：体外低能量震波 人脐静脉内皮细胞 血管内皮生长因子 白细胞介素-8 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学]