国际脑血管病杂志
國際腦血管病雜誌
국제뇌혈관병잡지
INTERNATIONAL JOURNAL OF CEREBROVASCULAR DISEASES
2012年
8期
595-600
,共6页
皮燕%张莉莉%黎炳护%高长越%王景周%王建红%胡子成%唐春花%郭露%李敬诚
皮燕%張莉莉%黎炳護%高長越%王景週%王建紅%鬍子成%唐春花%郭露%李敬誠
피연%장리리%려병호%고장월%왕경주%왕건홍%호자성%당춘화%곽로%리경성
NADPH氧化酶%T样受体4%炎症%肌,平滑,血管%动脉粥样硬化%细胞,培养的%小鼠
NADPH氧化酶%T樣受體4%炎癥%肌,平滑,血管%動脈粥樣硬化%細胞,培養的%小鼠
NADPH양화매%T양수체4%염증%기,평활,혈관%동맥죽양경화%세포,배양적%소서
NADPH Oxidase%Toll-Like Receptor 4%Inflammation%Muscle,Smooth,Vascular%Atherosclerosis%Cells,Cultured%Mice
目的 探讨NADPH氧化酶对培养的小鼠血管平滑肌细胞(vascular smooth muscle cell,VSMC)Toll样受体4(Toll-like receptor 4,TLR4)介导的炎症表型中的作用.方法 分别采用NADPH氧化酶激动剂血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)和抑制剂夹竹桃麻素(apocynin)处理培养的C57BL/6J和TLR4-/-小鼠胸主动脉VSMC.分别采用荧光探针二氯荧光素双醋酸盐染色法检测VSMC内活性氧(reactive oxygen species,ROS)含量,酶联免疫吸附法检测VSMC白细胞介素(interleukin,IL)-6、IL-1β和肿瘤坏死因子-α(tumor necrosis factor-o,TNF-α)表达,四甲基偶氮唑蓝染色法和Boyden小室检测VSMC的增殖和迁移.结果 PDGF-BB处理可显著增高C57BL/6J和TLR4-/-VSMC内ROS含量,而夹竹桃麻素可抑制ROS生成.PDGF-BB处理可使C57BL/6J VSMC IL-6[(52.69±3.49)ng/ml对(35.04±2.74) ng/ml,P=0.001]、IL-1β[(79.68±2.33) ng/ml对(62.38±0.54)ng/ml,P=0.000]和TNF-α[(218.35±5.42)ng/ml对(124.74±4.59) ng/ml,P=0.000]表达显著上调,增殖(1.69±0.53对1.04±0.40,P=0.000)和迁移(42.ll±4.05对1.69±0.53,P=0.000)能力均显著增强,而夹竹桃麻素预处理则可显著抑制VSMC IL-6[(42.11±4.05) ng/ml对(52.69±3.49) ng/ml,P=0.010]、IL-1β[(67.57± 1.36)ng/ml对(79.68±2.33) ng/ml,P=0.000]和TNF-α[(156.18±6.98) ng/ml对(218.35±5.42) ng/ml,P=0.000]表达以及增殖(1.23±0.42对1.69± 0.53,P=0.000)和迁移(42.11±4.05对52.69±3.49,P=0.000).TLR4-/-VSMC经PDGF-BB和夹竹桃麻素处理后,IL-6、IL-1β和TNF-α表达以及增殖和迁移能力均无显著变化.结论 NADPH氧化酶衍生的ROS参与了TLR4介导的VSMC炎症表型以及增殖和迁移,可能是其影响动脉粥样硬化发生和发展的重要机制.
目的 探討NADPH氧化酶對培養的小鼠血管平滑肌細胞(vascular smooth muscle cell,VSMC)Toll樣受體4(Toll-like receptor 4,TLR4)介導的炎癥錶型中的作用.方法 分彆採用NADPH氧化酶激動劑血小闆衍生生長因子-BB(platelet-derived growth factor-BB,PDGF-BB)和抑製劑夾竹桃痳素(apocynin)處理培養的C57BL/6J和TLR4-/-小鼠胸主動脈VSMC.分彆採用熒光探針二氯熒光素雙醋痠鹽染色法檢測VSMC內活性氧(reactive oxygen species,ROS)含量,酶聯免疫吸附法檢測VSMC白細胞介素(interleukin,IL)-6、IL-1β和腫瘤壞死因子-α(tumor necrosis factor-o,TNF-α)錶達,四甲基偶氮唑藍染色法和Boyden小室檢測VSMC的增殖和遷移.結果 PDGF-BB處理可顯著增高C57BL/6J和TLR4-/-VSMC內ROS含量,而夾竹桃痳素可抑製ROS生成.PDGF-BB處理可使C57BL/6J VSMC IL-6[(52.69±3.49)ng/ml對(35.04±2.74) ng/ml,P=0.001]、IL-1β[(79.68±2.33) ng/ml對(62.38±0.54)ng/ml,P=0.000]和TNF-α[(218.35±5.42)ng/ml對(124.74±4.59) ng/ml,P=0.000]錶達顯著上調,增殖(1.69±0.53對1.04±0.40,P=0.000)和遷移(42.ll±4.05對1.69±0.53,P=0.000)能力均顯著增彊,而夾竹桃痳素預處理則可顯著抑製VSMC IL-6[(42.11±4.05) ng/ml對(52.69±3.49) ng/ml,P=0.010]、IL-1β[(67.57± 1.36)ng/ml對(79.68±2.33) ng/ml,P=0.000]和TNF-α[(156.18±6.98) ng/ml對(218.35±5.42) ng/ml,P=0.000]錶達以及增殖(1.23±0.42對1.69± 0.53,P=0.000)和遷移(42.11±4.05對52.69±3.49,P=0.000).TLR4-/-VSMC經PDGF-BB和夾竹桃痳素處理後,IL-6、IL-1β和TNF-α錶達以及增殖和遷移能力均無顯著變化.結論 NADPH氧化酶衍生的ROS參與瞭TLR4介導的VSMC炎癥錶型以及增殖和遷移,可能是其影響動脈粥樣硬化髮生和髮展的重要機製.
목적 탐토NADPH양화매대배양적소서혈관평활기세포(vascular smooth muscle cell,VSMC)Toll양수체4(Toll-like receptor 4,TLR4)개도적염증표형중적작용.방법 분별채용NADPH양화매격동제혈소판연생생장인자-BB(platelet-derived growth factor-BB,PDGF-BB)화억제제협죽도마소(apocynin)처리배양적C57BL/6J화TLR4-/-소서흉주동맥VSMC.분별채용형광탐침이록형광소쌍작산염염색법검측VSMC내활성양(reactive oxygen species,ROS)함량,매련면역흡부법검측VSMC백세포개소(interleukin,IL)-6、IL-1β화종류배사인자-α(tumor necrosis factor-o,TNF-α)표체,사갑기우담서람염색법화Boyden소실검측VSMC적증식화천이.결과 PDGF-BB처리가현저증고C57BL/6J화TLR4-/-VSMC내ROS함량,이협죽도마소가억제ROS생성.PDGF-BB처리가사C57BL/6J VSMC IL-6[(52.69±3.49)ng/ml대(35.04±2.74) ng/ml,P=0.001]、IL-1β[(79.68±2.33) ng/ml대(62.38±0.54)ng/ml,P=0.000]화TNF-α[(218.35±5.42)ng/ml대(124.74±4.59) ng/ml,P=0.000]표체현저상조,증식(1.69±0.53대1.04±0.40,P=0.000)화천이(42.ll±4.05대1.69±0.53,P=0.000)능력균현저증강,이협죽도마소예처리칙가현저억제VSMC IL-6[(42.11±4.05) ng/ml대(52.69±3.49) ng/ml,P=0.010]、IL-1β[(67.57± 1.36)ng/ml대(79.68±2.33) ng/ml,P=0.000]화TNF-α[(156.18±6.98) ng/ml대(218.35±5.42) ng/ml,P=0.000]표체이급증식(1.23±0.42대1.69± 0.53,P=0.000)화천이(42.11±4.05대52.69±3.49,P=0.000).TLR4-/-VSMC경PDGF-BB화협죽도마소처리후,IL-6、IL-1β화TNF-α표체이급증식화천이능력균무현저변화.결론 NADPH양화매연생적ROS삼여료TLR4개도적VSMC염증표형이급증식화천이,가능시기영향동맥죽양경화발생화발전적중요궤제.
Objective To investigate the effect of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase on Toll-like receptor 4 (TLR4)-mediated proinflammatory phenotype of cultured vascular smooth muscle cells (VSMCs) in mice.Methods NADPH oxidase agonist platelet-derived growth factorBB (PDGF-BB) and inhibitor apocynin were used respectively to treat cultured VSMCs from C57BL/6J and TLR4-/-mice.The fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate was used to detect the reactive oxygen species (ROS) level in VSMCs.An enzyme-linked immunosorbent assay was used to detect the expressions of interleukin (IL)-6,IL-1β,and tumor necrosis factor-α (TNF-α) in VSMCs.Tetrazolium blue staining and Boyden chamber assay were used to detect the proliferation and migration of VSMC.Results The ROS levels were increased in VSMCs both from C57BL/6J and TLR4-/-mice after PDGF-BB treatment,and this could be inhibited by apocynin.PDGF-BB pretreatment significantly upregulated the expressions of IL-6 (52.69 ±3.49 ng/ml vs.35.04 ±2.74 ng/ml; P =0.001),IL-1β (79.68 ±2.33 ng/ml vs.62.38 ±0.54 ng/ml;P=0.000),and TNF-α (218.35± 5.42 ng/mlvs.124.74± 4.59 ng/ml; P=0.000) in VSMCs from C57BL/6J mice,and the abilities of proliferation (1.69 ± 0.53 vs.1.04 ± 0.40; P =0.000) and migration (42.11 ±4.05 vs.1.69 ± 0.53; P =0.000) were increased significantly; apocynin pretreatment significantly inhibit the expressions of IL-6 (42.11 ± 4.05 ng/ml vs.52.69 ± 3.49 ng/ml; P =0.010),IL-1β (67.57 ± 1.36 ng/ml vs.79.68 ±2.33 ng/ml; P =0.000) and TNF-α (156.18 ± 6.98 ng/ml vs.218.35 ± 5.42 ng/ml;P =0.000),as well as proliferation (1.23 ±0.42 vs.1.69 ±0.53; P =0.000) and migration (42.11 ±4.05 vs.52.69 ± 3.49; P =0.000).While there were no significant changes in the expressions of IL-6,IL-1β,and TNF-α in VSMCs from TLR4-/-mice after PDGF-BB and apocynin pretreatment.Conclusions NADPH oxidase-derived ROS involved in the TLR4-mediated VSMC inflammatory phenotype as well as proliferation and migration,which may be the important mechanisms of its influencing on the occurrence and development of atherosclerosis.
