中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
2期
139-143
,共5页
鄢高亮%胡圣大%王庆捷%王栋%王婧%乔勇%汤成春%马根山%顾雨春
鄢高亮%鬍聖大%王慶捷%王棟%王婧%喬勇%湯成春%馬根山%顧雨春
언고량%호골대%왕경첩%왕동%왕청%교용%탕성춘%마근산%고우춘
地高辛%高血压,肺性%缺氧%大鼠
地高辛%高血壓,肺性%缺氧%大鼠
지고신%고혈압,폐성%결양%대서
Digoxin%Hypertension,pulmonary%Hypoxia%Rats
目的 探讨地高辛早期干预对低氧诱导肺动脉高压(PAH)大鼠肺血管重构的影响及其机制.方法 48只SD大鼠按随机数字表法随机均分为4组并分别进行如下处理:(1)常氧对照组,(2)常氧+地高辛组,(3)低氧对照组,(4)低氧+地高辛组.常氧干预大鼠正常氧浓度(21％)仓内饲养,低氧干预大鼠则置于低氧仓内,氧浓度控制(10.5±0.5)％范围内,每日间断低氧8h(8:00～16:00);地高辛干预大鼠每次入仓前腹腔注射地高辛注射液1.0 mg·kg-1·d-1,对照组大鼠每次入仓前腹腔注射等体积注射用水.21 d后测定各组大鼠肺动脉平均压(mPAP),计算右室肥厚指数[RV/(LV+S)],测定管壁厚度占血管外径的百分比(WT％)和管壁面积占血管总面积的百分比(WA％).肺动脉平滑肌细胞培养采用组织贴块法,细胞增殖及迁移检测分别采用噻唑蓝比色法和Transwell小室法.肺动脉平滑肌细胞收缩表型标志物α肌动蛋白、调宁蛋白和肌动蛋白相关蛋白mRNA水平检测采用实时定量PCR法.肺动脉平滑肌细胞基质金属蛋白酶(MMPs)分泌水平检测采用Western印迹法.结果 低氧对照组大鼠mPAP、RV/(LV+S)、WT％、WA％均显著高于常氧对照组[(38.5±2.3)比(21.0±1.5)mmHg(1 mmHg =0.133 kPa)、(42.8±2.6)％比(26.7±1.5)％、(39.3±2.0)％比(17.5±3.3)％、(79.5±5.7)％比(45.7±4.9)％,均P＜0.05].而低氧+地高辛组大鼠上述指标均显著低于低氧对照组[(27.3±2.7)比(38.5 ±2.3)mmHg、(30.9±3.3)％比(42.8 ±2.6)％、(21.7±3.6)％比(39.3±2.0)％、(56.3±4.7)％比(79.5±5.7)％,均P＜0.05].地高辛(100 nmol/L)显著抑制低氧诱导的肺动脉平滑肌细胞增殖(吸光度值:0.575±0.030比0.747±0.039,P＜0.05),减少低氧诱导的肺动脉平滑肌细胞迁移31.9％ (P ＜0.05),并抑制低氧引起的收缩表型标志物mRNA表达的下降和MMP-2、MMP-9蛋白表达的升高.结论 地高辛早期应用可延缓低氧诱导的大鼠PAH形成及肺血管重构,该作用可能是通过抑制肺动脉平滑肌细胞增殖、迁移、表型转换和MMPs分泌而起作用.
目的 探討地高辛早期榦預對低氧誘導肺動脈高壓(PAH)大鼠肺血管重構的影響及其機製.方法 48隻SD大鼠按隨機數字錶法隨機均分為4組併分彆進行如下處理:(1)常氧對照組,(2)常氧+地高辛組,(3)低氧對照組,(4)低氧+地高辛組.常氧榦預大鼠正常氧濃度(21％)倉內飼養,低氧榦預大鼠則置于低氧倉內,氧濃度控製(10.5±0.5)％範圍內,每日間斷低氧8h(8:00～16:00);地高辛榦預大鼠每次入倉前腹腔註射地高辛註射液1.0 mg·kg-1·d-1,對照組大鼠每次入倉前腹腔註射等體積註射用水.21 d後測定各組大鼠肺動脈平均壓(mPAP),計算右室肥厚指數[RV/(LV+S)],測定管壁厚度佔血管外徑的百分比(WT％)和管壁麵積佔血管總麵積的百分比(WA％).肺動脈平滑肌細胞培養採用組織貼塊法,細胞增殖及遷移檢測分彆採用噻唑藍比色法和Transwell小室法.肺動脈平滑肌細胞收縮錶型標誌物α肌動蛋白、調寧蛋白和肌動蛋白相關蛋白mRNA水平檢測採用實時定量PCR法.肺動脈平滑肌細胞基質金屬蛋白酶(MMPs)分泌水平檢測採用Western印跡法.結果 低氧對照組大鼠mPAP、RV/(LV+S)、WT％、WA％均顯著高于常氧對照組[(38.5±2.3)比(21.0±1.5)mmHg(1 mmHg =0.133 kPa)、(42.8±2.6)％比(26.7±1.5)％、(39.3±2.0)％比(17.5±3.3)％、(79.5±5.7)％比(45.7±4.9)％,均P＜0.05].而低氧+地高辛組大鼠上述指標均顯著低于低氧對照組[(27.3±2.7)比(38.5 ±2.3)mmHg、(30.9±3.3)％比(42.8 ±2.6)％、(21.7±3.6)％比(39.3±2.0)％、(56.3±4.7)％比(79.5±5.7)％,均P＜0.05].地高辛(100 nmol/L)顯著抑製低氧誘導的肺動脈平滑肌細胞增殖(吸光度值:0.575±0.030比0.747±0.039,P＜0.05),減少低氧誘導的肺動脈平滑肌細胞遷移31.9％ (P ＜0.05),併抑製低氧引起的收縮錶型標誌物mRNA錶達的下降和MMP-2、MMP-9蛋白錶達的升高.結論 地高辛早期應用可延緩低氧誘導的大鼠PAH形成及肺血管重構,該作用可能是通過抑製肺動脈平滑肌細胞增殖、遷移、錶型轉換和MMPs分泌而起作用.
목적 탐토지고신조기간예대저양유도폐동맥고압(PAH)대서폐혈관중구적영향급기궤제.방법 48지SD대서안수궤수자표법수궤균분위4조병분별진행여하처리:(1)상양대조조,(2)상양+지고신조,(3)저양대조조,(4)저양+지고신조.상양간예대서정상양농도(21％)창내사양,저양간예대서칙치우저양창내,양농도공제(10.5±0.5)％범위내,매일간단저양8h(8:00～16:00);지고신간예대서매차입창전복강주사지고신주사액1.0 mg·kg-1·d-1,대조조대서매차입창전복강주사등체적주사용수.21 d후측정각조대서폐동맥평균압(mPAP),계산우실비후지수[RV/(LV+S)],측정관벽후도점혈관외경적백분비(WT％)화관벽면적점혈관총면적적백분비(WA％).폐동맥평활기세포배양채용조직첩괴법,세포증식급천이검측분별채용새서람비색법화Transwell소실법.폐동맥평활기세포수축표형표지물α기동단백、조저단백화기동단백상관단백mRNA수평검측채용실시정량PCR법.폐동맥평활기세포기질금속단백매(MMPs)분비수평검측채용Western인적법.결과 저양대조조대서mPAP、RV/(LV+S)、WT％、WA％균현저고우상양대조조[(38.5±2.3)비(21.0±1.5)mmHg(1 mmHg =0.133 kPa)、(42.8±2.6)％비(26.7±1.5)％、(39.3±2.0)％비(17.5±3.3)％、(79.5±5.7)％비(45.7±4.9)％,균P＜0.05].이저양+지고신조대서상술지표균현저저우저양대조조[(27.3±2.7)비(38.5 ±2.3)mmHg、(30.9±3.3)％비(42.8 ±2.6)％、(21.7±3.6)％비(39.3±2.0)％、(56.3±4.7)％비(79.5±5.7)％,균P＜0.05].지고신(100 nmol/L)현저억제저양유도적폐동맥평활기세포증식(흡광도치:0.575±0.030비0.747±0.039,P＜0.05),감소저양유도적폐동맥평활기세포천이31.9％ (P ＜0.05),병억제저양인기적수축표형표지물mRNA표체적하강화MMP-2、MMP-9단백표체적승고.결론 지고신조기응용가연완저양유도적대서PAH형성급폐혈관중구,해작용가능시통과억제폐동맥평활기세포증식、천이、표형전환화MMPs분비이기작용.
Objective To explore the effects of digoxin on hypoxia-induced pulmonary artery hypertension (PAH) and the possible mechanisms.Methods A total of 48 Sprague-Dawley rats were randomly divided into 4 groups:normoxia control,normoxia + digoxin,hypoxia control and hypoxia +digoxin.The animals were exposed to chronic intermittent hypoxia (PO2:(10.5 ±0.5) ％,8:00-16:00)or room air for 21 days.Each rat received a daily intraperitoneal injection of either digoxin (1.0 mg· kg-1 ·d-1) or an equal volume of vehicle,starting at the first day of hypoxia or normoxia.At Day 21,mean pulmonary arterial pressure (mPAP),right ventricular hypertrophy (RV/(LV + S)) and index of wall thickness of small pulmonary artery (WT％ and WA％) among groups were compared.And in vitro the changes of pulmonary artery smooth muscle cells (PASMCs) proliferation were determined by methyl thiazolyl tetrazolium (MTT) assay.Migration assay was performed with a Transwell chamber.Real-time quantitative polymerase chain reaction (PCR) was performed to quantify the mRNA levels of smooth muscle cell phenotype markers such as smooth muscle-α-actin,calponin and smooth muscle 22α under normoxic or hypoxic conditions in the absence or presence of digoxin.And the protein expressions of matrix metalloproteinase (MMPs)were determined by Western blot.Results Digoxin treatment significantly lowered mPAP,reduced WT％ and WA％ and right ventricular hypertrophy compared with those of the hypoxic group (mPAP:(27.3 ± 2.7) vs (38.5 ± 2.3) mmHg(1 mmHg =0.133 kPa); RV/(LV + S):(30.9±3.3)％ vs (42.8±2.6)％,WT％:(21.7±3.6)％ vs (39.3±2.0)％; WA％:(56.3±4.7)％ vs (79.5 ±5.7)％,all P ＜0.05).And in vitro,digoxin restored the hypoxia-induced inhibition of the expression of smooth muscle cell phenotype markers and prevented the hypoxia-induced activation of MMPs in PASMCs.Conclusion Early digoxin therapy reduces pulmonary artery remodeling in hypoxiainduced PAH rat model and this effect is probably correlated with the inhibitions of proliferation,migration,phenotype switching and expression of MMPs induced by hypoxia in PASMCs.
