国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2012年
9期
597-602
,共6页
申艳%李冰冰%杨许丽%朱杰%梁樱%马正良
申豔%李冰冰%楊許麗%硃傑%樑櫻%馬正良
신염%리빙빙%양허려%주걸%량앵%마정량
二氯乙酸盐%肺动脉高压%存活素
二氯乙痠鹽%肺動脈高壓%存活素
이록을산염%폐동맥고압%존활소
Dichloroacetate%Pulmonary arterial hypertension%Survivin
目的 观察早期和晚期二氯乙酸盐(dichloroacetate,DCA)干预对左肺切除(pneumonectomy,PE)联合野百合碱(monocrotaline,MCT)皮下注射诱导大鼠肺动脉高压(pulmonary arterial hypertension,PAH)的治疗作用以及对生存素(survivin)表达的影响.方法 雄性SD大鼠(体重200 g～250 g)采用随机数字表法随机分为5组,每组8只:正常对照组(N),DCA对照组(D),肺高压模型组(PM),早期DC预防组(PMD1),晚期DCA治疗组(PMD2).除N组和D组外,其余组别大鼠均行左肺切除,并于14 d后项背部皮下注射MCT(剂量为60 mg/kg).同时PMD1组和PMD2组分别在左肺切除14 d和35 d后开始每天给予DCA(剂量为80 mg· kg-1·d-1)灌胃,而PM组在左肺切除14 d后给予相同容积的生理盐水灌胃,D组同N组大鼠,只是左肺切除14 d后同样给予相同剂量的DCA灌胃.PMD1组和PMD2组大鼠分别在左肺切除35 d和56 d行开胸进行肺动脉压力(PAP)的测量.测量后处死动物,取肺组织进行常规病理染色,采用荧光定量PCR法检测survivin mRNA的表达,采用western blot法检测大鼠survivin蛋白的表达.结果 PM组大鼠平均PAP为(37.8±1.3) mm Hg(1 mm Hg=0.133 kPa),肺小血管中膜厚度占外径的百分比(WT％)和右心室与左心室加室间隔之比(RV/LV+S)分别为(40±8)％和(44.8±2.2),survivin mRNA和Western blot蛋白表达分别为(4.03±0.27)和(0.684±0.055),与N组、D组比较均明显升高(P＜0.05).早期DCA预防后PAP明显下降为(26.2±1.5) mm Hg,WT％和RV/LV+S分别降为(24±3)％和(29.5±1.9),而survivin mRNA和western blot蛋白表达也分别降为(2.00±0.12)和(0.415±0.027),与PM组比较均明显下降(P＜0.05),而晚期干预后PAP为(36.0±1.6) mm Hg,WT％和RV/LV+S分别为(38±3)％和(42.0±1.3),survivin mRNA和western blot蛋白表达也分别(3.77±0.45)和(0.732±0.101),与PM组比较没有统计学差异(P＞0.05).结论 Survivin在PAH的发生及肺血管重构中起重要作用,而DCA早期预防能缓解PAP升高和肺血管中膜增厚以及降低survivin的表达量,然而晚期干预后和PM组比较差异没有统计学意义.
目的 觀察早期和晚期二氯乙痠鹽(dichloroacetate,DCA)榦預對左肺切除(pneumonectomy,PE)聯閤野百閤堿(monocrotaline,MCT)皮下註射誘導大鼠肺動脈高壓(pulmonary arterial hypertension,PAH)的治療作用以及對生存素(survivin)錶達的影響.方法 雄性SD大鼠(體重200 g～250 g)採用隨機數字錶法隨機分為5組,每組8隻:正常對照組(N),DCA對照組(D),肺高壓模型組(PM),早期DC預防組(PMD1),晚期DCA治療組(PMD2).除N組和D組外,其餘組彆大鼠均行左肺切除,併于14 d後項揹部皮下註射MCT(劑量為60 mg/kg).同時PMD1組和PMD2組分彆在左肺切除14 d和35 d後開始每天給予DCA(劑量為80 mg· kg-1·d-1)灌胃,而PM組在左肺切除14 d後給予相同容積的生理鹽水灌胃,D組同N組大鼠,隻是左肺切除14 d後同樣給予相同劑量的DCA灌胃.PMD1組和PMD2組大鼠分彆在左肺切除35 d和56 d行開胸進行肺動脈壓力(PAP)的測量.測量後處死動物,取肺組織進行常規病理染色,採用熒光定量PCR法檢測survivin mRNA的錶達,採用western blot法檢測大鼠survivin蛋白的錶達.結果 PM組大鼠平均PAP為(37.8±1.3) mm Hg(1 mm Hg=0.133 kPa),肺小血管中膜厚度佔外徑的百分比(WT％)和右心室與左心室加室間隔之比(RV/LV+S)分彆為(40±8)％和(44.8±2.2),survivin mRNA和Western blot蛋白錶達分彆為(4.03±0.27)和(0.684±0.055),與N組、D組比較均明顯升高(P＜0.05).早期DCA預防後PAP明顯下降為(26.2±1.5) mm Hg,WT％和RV/LV+S分彆降為(24±3)％和(29.5±1.9),而survivin mRNA和western blot蛋白錶達也分彆降為(2.00±0.12)和(0.415±0.027),與PM組比較均明顯下降(P＜0.05),而晚期榦預後PAP為(36.0±1.6) mm Hg,WT％和RV/LV+S分彆為(38±3)％和(42.0±1.3),survivin mRNA和western blot蛋白錶達也分彆(3.77±0.45)和(0.732±0.101),與PM組比較沒有統計學差異(P＞0.05).結論 Survivin在PAH的髮生及肺血管重構中起重要作用,而DCA早期預防能緩解PAP升高和肺血管中膜增厚以及降低survivin的錶達量,然而晚期榦預後和PM組比較差異沒有統計學意義.
목적 관찰조기화만기이록을산염(dichloroacetate,DCA)간예대좌폐절제(pneumonectomy,PE)연합야백합감(monocrotaline,MCT)피하주사유도대서폐동맥고압(pulmonary arterial hypertension,PAH)적치료작용이급대생존소(survivin)표체적영향.방법 웅성SD대서(체중200 g～250 g)채용수궤수자표법수궤분위5조,매조8지:정상대조조(N),DCA대조조(D),폐고압모형조(PM),조기DC예방조(PMD1),만기DCA치료조(PMD2).제N조화D조외,기여조별대서균행좌폐절제,병우14 d후항배부피하주사MCT(제량위60 mg/kg).동시PMD1조화PMD2조분별재좌폐절제14 d화35 d후개시매천급여DCA(제량위80 mg· kg-1·d-1)관위,이PM조재좌폐절제14 d후급여상동용적적생리염수관위,D조동N조대서,지시좌폐절제14 d후동양급여상동제량적DCA관위.PMD1조화PMD2조대서분별재좌폐절제35 d화56 d행개흉진행폐동맥압력(PAP)적측량.측량후처사동물,취폐조직진행상규병리염색,채용형광정량PCR법검측survivin mRNA적표체,채용western blot법검측대서survivin단백적표체.결과 PM조대서평균PAP위(37.8±1.3) mm Hg(1 mm Hg=0.133 kPa),폐소혈관중막후도점외경적백분비(WT％)화우심실여좌심실가실간격지비(RV/LV+S)분별위(40±8)％화(44.8±2.2),survivin mRNA화Western blot단백표체분별위(4.03±0.27)화(0.684±0.055),여N조、D조비교균명현승고(P＜0.05).조기DCA예방후PAP명현하강위(26.2±1.5) mm Hg,WT％화RV/LV+S분별강위(24±3)％화(29.5±1.9),이survivin mRNA화western blot단백표체야분별강위(2.00±0.12)화(0.415±0.027),여PM조비교균명현하강(P＜0.05),이만기간예후PAP위(36.0±1.6) mm Hg,WT％화RV/LV+S분별위(38±3)％화(42.0±1.3),survivin mRNA화western blot단백표체야분별(3.77±0.45)화(0.732±0.101),여PM조비교몰유통계학차이(P＞0.05).결론 Survivin재PAH적발생급폐혈관중구중기중요작용,이DCA조기예방능완해PAP승고화폐혈관중막증후이급강저survivin적표체량,연이만기간예후화PM조비교차이몰유통계학의의.
Objective To observe the effect of dichloroacetate (DCA) on pulmonary arterial pressure and the expression of survivin in rat model of severe pulmonary arterial hypertention(PAH).Methods 40 male Sprague-Dawley rats (8-wk-old,weight 200 g-250 g) were randomly divided into 5 goups,each of which had eight rats:normal group (N),dichloroacetate control group (D),left pneumonectom (PE)+ monocrotaline (MCT)group (PM),early DCA treated group (PMD1),and later DCA treated group (PMD2).Except for group N and D,other groups were induced by pneumonectomy combined with monocrotaline 60 mg/kg subcutaneous injection after pneumonectomy(D14).Group PMD1 and PMD2 were respectively received dichloroacetate 80 mg· kg-1·d-1 after pneumonectomy (D14) and D35 by intragastric gavage once a day for 21 consecutive days.At the same time,the equivalent volume of sterile saline solution was substituted for DCA in group PM.Rats in group D were treated as same as in group N except for receiving DCA gavage treatment as described in group PMD.At D35 (PMD1)and D56 (PMD2).Pulmonary arterial pressure was measured by direct puncture of pulmonary artery trunk with i.v.Catheter (22 gauge) connected to pressure transducer.Rats were euthanized by intravenous injection with potassium chloride,and inferior lobe of right lung were dissected,snap-frozen in the liquid nitrogen and kept at-80 ℃ until analyzed.The residual lobes were fixed in neutral formaldehyde for further HE stain and morphometric analysis.The mRNA expression of survivin was analyzed with realtime-PCR.The protein expression of survivin was detected with Western blot.Results The mean PAP of group PM is (37.8±1.3) mm Hg (1 mm Hg=0.133 kPa),the percentage of medial thickening (WT％) and right ventricular hypertrophy index (RV/LV+S) are (40±8)％ and (44.8±2.2),respectively.The expression of survivin mRNA and protein level are(4.03±0.27) and(0.684±0.055),respectively.Compared with group N and group D,they were markedly increased in group PM (P＜0.05).Early treatment with DCA significandy reduced the mean PAP,the percentage of medial thickening,and right ventricular hypertrophy index,they are (26.2±1.5) mm Hg,(24±3)％,and (29.5±1.9),respectively.Meanwhile,the expression of survivin mRNA and protein level were markedly reduced to (2.00±0.12) and (0.415±0.027),compared with group PM,they were significantly reduced in group PMD1 (P＜0.05).Whereas later treatment with DCA has no significant effect (P＞0.05),the mean PAP,the percentage of medial thickening,and right ventricular hypertrophy index are (36.0± 1.6) mm Hg,(38±3)％,and (42.0±1.3),respectively.The expression of survivin mRNA and protein level were (3.77±0.45) and (0.732±0.101),respectively.Conclusions Survivin plays an important role in the development of pulmonary hypertention and the remodeding of pulmonary vascular.DCA alleviates the deterioration of PAH,medial thickening of pulmonary arterioles and protein expression of survivin in pneumonectomized rats injected with MCT,whereas there were no significant difference was found between PM and PMD2.
