当归补血汤在流体剪切应力环境中对内皮细胞功能的影响

Effects of Danggui Buxue Tang on endothelial cells exposed to flow shear stress

目的观察当归补血汤在流体剪切应力(FSS)环境中对人内皮细胞功能的影响。方法将细胞置于平行平板流动小室,加载稳定层流FSS(0,0.12,1.2,2.4 Pa)6 h后,将不同FSS环境中的细胞随机分为5组:对照组加入无血清培养液灌流;辛伐他汀组加入含0.1μmol/L辛伐他汀培养液灌流;3个中药组分别加入含3 g/L的当归补血汤(当归、黄芪比为1∶1、1∶3、1∶5)培养液灌流。以上各组经FSS加载12 h后,收集细胞及灌流液,检测细胞增殖力、一氧化氮(NO)含量,细胞内皮型一氧化氮合酶(eNOS)和血管内皮细胞生长因子受体2(VEGFR2)mRNA及蛋白的表达。结果对照组FSS为1.2、2.4 Pa时,内皮细胞表达eNOS、VEGFR2的水平较其在0、0.12 Pa时升高(P<0.05);与对照组相比,当归补血汤在FSS为0、0.12 Pa时能上调细胞eNOS、VEGFR2表达并增强其增殖和分泌NO功能(P<0.05)。且当归补血汤(当归、黄芪比为1∶3、1∶5)在FSS为1.2、2.4 Pa时可上调细胞eNOS、VEGFR2表达并增强其增殖和分泌NO功能(P<0.05)。结论 FSS环境是影响当归补血汤对内皮细胞作用效果的重要因素之一。

Objective To investigate the effects of Danggui Buxue Tang（ DGBX,angelica sinensis blood-tonifying decoction） on human endothelial cells in a steady flow shear stress（ FSS）. Methods Cells were incubated on a parallel plate flow chamber at a steady FSS of 0,0. 12,1. 2,and 2. 4 Pa for 6 hours. Cells exposed to different FSS were randomly divided into 5 groups： control group perfused with M199 culture medium,simvastatin group（ 0. 1 μmol/L simvastatin）,and DGBX groups { 3 g/L DGBX decoction： Chinese angelica（ Danggui,Radix Angelica Sinensis,AS） and astragalus root（ Huangqi,Radix Astragali,RA） at the dose ratios of 1 ∶1,1 ∶3,and 1 ∶5}. After 12 hours,the cellular proliferation,levels of nitric oxide（ NO）,expressions of endothelial nitric oxide synthase（ eNOS）,vascular endothelial growth factor receptor 2（ VEGFR2） mRNAs and proteins were measured. Results In control group,eNOS and VEGFR2 levels in the endothelial cells exposed to FSS at 1. 2 and 2. 4 Pa were significantly higher than those of cells exposed to FSS at 0 and 0. 12 Pa（ P 〈 0. 05）. Compared with the control group,DGBX decoction up-regulated the cellular proliferation and expression of eNOS and VEGFR2,and NO levels at FSS of 0 and 0. 12 Pa（ P 〈 0. 05）. And DGBX decoction（ AS∶ RA at 1∶3,1∶5） up-regulated the expressions of eNOS and VEGFR2,cellular proliferation and NO level at FSS of 1. 2,2. 4 Pa（ P 0. 05）. Conclusion FSS seems to play an important role in the effects of DGBX decoction on endothelial cells.