CAJ | 학술논문

目的：氧化低密度脂蛋白（ oxidized low density lipoprotein ，ox-LDL）损伤血管内皮细胞是动脉粥样硬化的重要起始环节之一。文中旨在观察通心络对ox-LDL诱导损伤的血管内皮细胞的保护作用。方法采用ox-LDL （终浓度为30 mg／L）造成血管内皮细胞氧化应激损伤模型，将人脐静脉内皮细胞（ human umbilical vein endothelial cell ，HUVEC）分为正常对照组、模型组、通心络高、中、低剂量组，正常对照组给予无血清DMEM培养基培养，模型组给予含ox-LDL（终浓度为30 mg／L）的无血清DMEM培养基培养24 h，通心络低、中、高剂量组给予含不同浓度通心络（终浓度为50、100、150 mg／L）预培养4 h后加入ox-LDL（终浓度为30 mg／L）继续培养24 h。 MTS法检测各组细胞的生存活性；试剂盒检测各组细胞上清液中NO含量、SOD的活力和线粒体膜电位变化；Western blot法检测各组细胞诱导型NO合酶（ inducible nitric oxide synthase ，iNOS）、基质金属蛋白酶9（ matrix metalloproteinase9，MMP9）、核因子κB（ nuclear factor kappa B ，NF-κB） p65蛋白表达情况。结果与正常对照组比较，模型组HUVEC生存活性明显降低[（100．00±2．23）％ vs （73．89±0．67）％， P＜0．01]，与模型组比较，通心络中、高剂量组HUVECs生存活性[（92．15±0．76）％、（97．19±1．45）％]明显提高（P＜0．01）。与正常对照组细胞线粒体膜电位、细胞培养液中NO含量、SOD活力比较，模型组明显降低（P＜0．01）；而通心络低、中、高剂量组较模型组明显升高（P＜0．01）。与正常对照组iNOS、MMP9、NF-κBp65蛋白表达比较，模型组明显升高（P＜0．01）；而通心络低、中，高剂量组较模型组明显降低（P＜0．05）。结论通心络有较强的抗氧化、抗炎能力，可减轻ox-LDL对血管内皮细胞的损伤。
목적：양화저밀도지단백（ oxidized low density lipoprotein ，ox-LDL）손상혈관내피세포시동맥죽양경화적중요기시배절지일。문중지재관찰통심락대ox-LDL유도손상적혈관내피세포적보호작용。방법채용ox-LDL （종농도위30 mg／L）조성혈관내피세포양화응격손상모형，장인제정맥내피세포（ human umbilical vein endothelial cell ，HUVEC）분위정상대조조、모형조、통심락고、중、저제량조，정상대조조급여무혈청DMEM배양기배양，모형조급여함ox-LDL（종농도위30 mg／L）적무혈청DMEM배양기배양24 h，통심락저、중、고제량조급여함불동농도통심락（종농도위50、100、150 mg／L）예배양4 h후가입ox-LDL（종농도위30 mg／L）계속배양24 h。 MTS법검측각조세포적생존활성；시제합검측각조세포상청액중NO함량、SOD적활력화선립체막전위변화；Western blot법검측각조세포유도형NO합매（ inducible nitric oxide synthase ，iNOS）、기질금속단백매9（ matrix metalloproteinase9，MMP9）、핵인자κB（ nuclear factor kappa B ，NF-κB） p65단백표체정황。결과여정상대조조비교，모형조HUVEC생존활성명현강저[（100．00±2．23）％ vs （73．89±0．67）％， P＜0．01]，여모형조비교，통심락중、고제량조HUVECs생존활성[（92．15±0．76）％、（97．19±1．45）％]명현제고（P＜0．01）。여정상대조조세포선립체막전위、세포배양액중NO함량、SOD활력비교，모형조명현강저（P＜0．01）；이통심락저、중、고제량조교모형조명현승고（P＜0．01）。여정상대조조iNOS、MMP9、NF-κBp65단백표체비교，모형조명현승고（P＜0．01）；이통심락저、중，고제량조교모형조명현강저（P＜0．05）。결론통심락유교강적항양화、항염능력，가감경ox-LDL대혈관내피세포적손상。
Ob jectiev Oxidized low-density lipoprotein ( ox-LDL) induces vascular endothelial cell injury , which is one of the factors initiating atherosclerosis .This study aimed to investigate the protective effect of Tongxinluo ( TXL ) on vascular endothelial cells with ox-LDL-induced injury . Methods Human umbilical vein endothelial cells ( HUVEC ) were cultured in vitro and divided into five groups:normal control, oxidative stress injury (OSI) model, and high, medium and low dose TXL.The HUVECs were incubated with ox-LDL at the concentration of 30 mg/L for 24 hours to induce oxidative stress injury and then treated with TXL at 50, 100 and 150 mg/L for 4 hours, followed by 24 hour incubation with 30 mg/L ox-LDL added to the culture medium .The viability of the cells was detected by MTS assay, the nitric oxide (NO) content, superoxide dismutase (SOD) activity and mitochondrial membrane poten-tial ( MMP) in the cell culture supernatant were measured with respective kits , and the expressions of iNOS , MMP9, and NF-κBp65 proteins were determined by Western blot . Results The HUVECs of the OSI model group showed a significant decrease in cell via-bility compared with the normal control , ([73 .89 ±0.67] vs [100.00 ±2.23]%, P<0.01) but a remarkably increase after treated with medium and high dose TXL ([92.15 ±0.76]%and [ 97.19 ±1.45]%, P<0.01).The MMP, NO content, and SOD activity were markedly reduced in the model group (P<0.01) but elevated in the low, medium, and high dose TXL groups (P<0 .01).The expressions of the iNOS, MMP9, and NF-κBp65proteins were significantly up -regulated in the model group (P<0.01) but down reg-ulated in the low, medium, and high dose TXL groups (P<0.05).C on clusion TXL has the effects of anti-oxidation and anti-in-flammation and can protect vascular endothelial cells against ox-LDL-induced injury .