CAJ | 학술논문

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 mJ／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（vas-cular 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、0．030、．09、0．18、0．24 mJ／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 mJ／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 mJ／mm2能量A450值与对照组比，差异有统计学意义（0．70±0．04 vs 0．54±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0．09mJ ／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 mJ／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。
목적：체외진파치료능촉진결혈조직적혈관신생화가속측지순배건립，단기구체작용궤제상불명학。탐토0．09 mJ／mm2적체외진파대인제정맥내피세포（human umbilical vein endothelial cells ， HUVECs）증식급혈관내피생장인자（vas-cular endothelial growth factor ， VEGF）、백세포개소-8（interleukin-8， IL-8）변화적영향。방법실험분위실험조화대조조，대조조시관치우진파의기중단불진행진파처리，실험조이0、0．030、．09、0．18、0．24 mJ／mm2진파능량충격，매공가CCK8용액，용매표의측정재파장450 nm처적A치。대체외배양적HUVECs세포주시여0．09 mJ／mm2능량적진파처리，CCK8비색법검측세포증식정황，Real-time PCR법화류식세포술분별검측저능량초성파처리후세포인자VEGF、IL-8 mRNA표체화단백질적변화。결과①CCK8비색법검측결과현시실험조중지유0．09 mJ／mm2능량A450치여대조조비，차이유통계학의의（0．70±0．04 vs 0．54±0．09， P＜0．05），이기타능량여대조조비，차이무통계학의의（P＞0．05）；②RT-PCR결과현시0．09mJ ／mm2능량시HUVECs VEGF、IL-8 mRNA적표체（7．93±0．90、7．34±1．67）균교대조조（1．07±0．40、1．00±0．09）명현승고（P＜0．01）。③류식세포술검측결과현시여대조조상비，0．09 mJ／mm2능량시HUVECs VEGF、IL-8단백표체（39．89±4．79、31．33±5．61）균교대조조（20．98±3．30、22．60±3．76）명현승고（P ＜0．05）。결론체외저능량진파가촉진HUVECs증식화증가VEGF、IL-8적표체， VEGF、IL-8적표체증가가재체외진파촉진혈관신생궤제중기중요작용。
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 flow cytometry 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 ).②RT-PCR showed that the 0.09 mJ/mm2 energy significantly enhanced the expressions of VEGF and IL-8 mRNA compared with the control group (7.93 ±0.90 vs 1.07 ±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/mm2 ESWT 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-regulation of VEGF and IL-8 may play an important role in the promotion of angiogenesis by ESWT .