CAJ | 학술논문

目的探讨五甲基槲皮素(PMQ)在体外和体内的抗静脉移植物狭窄作用.方法体外实验中用血管紧张素Ⅱ(AngⅡ)(0.1μmol/L,24 h)刺激血管平滑肌细胞(VSMC)增殖,同时分别给予0.1、0.3、1.0、3.0、10.0、30.0 μmol/L的PMQ干预,用MTT法测细胞活力,用DCFH-DA测活性氧簇(ROS),用real-time PCR测还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚单位Nox1,p47phox,p22phox的mRNA表达.体内实验中建立大鼠颈部自体静脉移植模型,随机分为模型对照组和给药组.对照组每晨溶剂灌胃,给药组每晨按PMQ 12.5、25.0、50.0mg/kg 3个剂量分别灌胃.于28天后取材测静脉移植物新生内膜和中膜的厚度及面积比.结果 PMQ 0.3 μmol/L开始显著抑制VSMC的活力,减少ROS的产生并显著抑制Nox1、p47phox、p22phox表达,3μmol/L时作用最强.与对照组相比,3个剂量组的PMQ均能显著减小内膜和中膜的厚度及面积比.结论 PMQ有抑制静脉移植物新生内膜增生的作用,可能与其抑制ROS产生和NADPH氧化酶表达有关.
목적 탐토오갑기곡피소(PMQ)재체외화체내적항정맥이식물협착작용.방법 체외실험중용혈관긴장소Ⅱ(AngⅡ)(0.1μmol/L,24 h)자격혈관평활기세포(VSMC)증식,동시분별급여0.1、0.3、1.0、3.0、10.0、30.0 μmol/L적PMQ간예,용MTT법측세포활력,용DCFH-DA측활성양족(ROS),용real-time PCR측환원형연선알선표령이핵감산린산(NADPH)양화매아단위Nox1,p47phox,p22phox적mRNA표체.체내실험중건립대서경부자체정맥이식모형,수궤분위모형대조조화급약조.대조조매신용제관위,급약조매신안PMQ 12.5、25.0、50.0mg/kg 3개제량분별관위.우28천후취재측정맥이식물신생내막화중막적후도급면적비.결과 PMQ 0.3 μmol/L개시현저억제VSMC적활력,감소ROS적산생병현저억제Nox1、p47phox、p22phox표체,3μmol/L시작용최강.여대조조상비,3개제량조적PMQ균능현저감소내막화중막적후도급면적비.결론 PMQ유억제정맥이식물신생내막증생적작용,가능여기억제ROS산생화NADPH양화매표체유관.
Objective Pentamethylquercetin (PMQ) has a role in cardiovascular protection. We investigate the effects of 3,3' ,4' ,5,7-pentamethylquercetin, a derivative of PMQ, on intimal hyperplasia of the vein grafts in rats both in vivo and in vitro. Methods The proliferation of vascular smooth muscle cells ( VSMC ) was induced with Ang Ⅱ (0. 1μmol/L, 24 h)while PMQ was administrated at six different dosages (0. 1, 0.3, 1,3, 10 and 30 μmoL/L). Cell viability was identified with MTT; ROS was measured with DCFH-DA; and the expression of NADPH oxidase subunits Nox1, p47phox, and p22phox mRNA were measured with real-time PCR. For the experiment in vivo, 24 SD rats were randomly assigned to control group and PMQ groups, the latter was further divided into three different dosage groups. In the control group, solvent was administrated daily via gavage. In PMQ groups, PMQ ( 12.5 mg/kg, 25 mg/kg, 50 mg/kg) was administrated daily respectively in the same way.All SD rats received operation performed by one person. Reversed external jugular vein was implanted into the external carotid of the same side with interrupted suture. 4 weeks after operation, all vein grafts were harvested. Status of the vein grafts was observed and tissue sections were analyzed with HE staining. The intimal hyperplasia ( intima/media area index and intima/media thickness index) of the vein grafts was assessed. Results Cell viability and ROS of VSMC induced by Ang Ⅱ were suppressed by PMQ. Cell viability and ROS of VSMC were increased substantially when treated with Ang Ⅱ. The therapeutic effects of PMQ could be initially identified at dose of0. 3 μmol/L, with a peak at 3 μmol/L. The effects decreased from 30μmol/L to 10 μmol/L. PMQ at dose of 0.1 μmol/L had no effect on cell viability and ROS of VSMC induced by Ang Ⅱ. PMQ also downregulated the mRNA expression of NADPH oxidase subunits Nox1, p47phox and p22phox induced by Ang Ⅱ. A peak effect was observed at 3μmoL/L and decreased at 30 μmol/L. PMQ at o. 1 μmol/L had no effect on mRNA expression of NADPH oxidase subunits induced by Ang Ⅱ. As compared with control group, PMQ decreased intima/media area index ( 1. 64 ±0.20 in control, 0. 74 ±0.18 at 12.5 mg/kg, 1.09 ±0.17 at 25 mg/kg, 1.21 ± 0. 21 at 50 mg/kg) and intima/media thickness index ( 1.34 ± 0. 24 in control, 0.67 ± 0. 17 at 12.5 mg/kg, 0. 74 ± 0.14 at 25 mg/kg, 0.93 ± 0. 18 at 50mg/kg) at three dosages after implantation. Conclusion PMQ may suppress the proliferation of VSMC and inhibit neointima hyperplasia of vein grafts in rats. The effects may be attributed to the anti-oxidative activity and the downregulation of mRNA expression of NADPH oxidase subunits Noxl, p47phox and p22phox.