实验性肺动脉高压大鼠心室间糖酵解代谢相关基因筛选和差异
실험성폐동맥고압대서심실간당효해대사상관기인사선화차이
Glycoltic metabolism gene changes in left and right ventricles in experimental rat pulmonary arterial hypertension model
  • 万方数据
    中华心血管病杂志 중화심혈관병잡지
    Chinese Journal of Cardiology
    2014年 12期 1010-1016 ,共7页

    邱梅红%张锐%郑杨%张尉华

    구매홍%장예%정양%장위화

    高血压,肺性%糖酵解 高血壓,肺性%糖酵解
    고혈압,폐성%당효해
    Hypertension,pulmonary%Glycolysis
    目的 探讨野百合碱(MCT)所致肺动脉高压(PAH)大鼠左、右心室能量代谢转移及可能的差异位点.方法 成年Sprague-Dawley大鼠,颈背部皮下一次性注射MCT(50 mg/kg)建立PAH右心衰竭模型,同时设立对照组,模型组按MCT作用时间又分为3组,即MCT-2周组、MCT-3周组和MCT-4周组(MCT-2w组、MCT-3w组和MCT-4w组).采用右心导管法检测各组大鼠血液动力学指标,右心室/(左心室+室间隔)重量计算右心室肥厚指数,免疫组织化学法检测心室壁细胞增殖细胞核抗原(PCNA)以及原位末端转移酶标记(TUNEL)的变化,实时定量PCR方法从糖酵解途径候选位点中筛选表达差异基因.结果 (1)血液动力学指标及右心室肥厚指数:MCT-3w组大鼠平均肺动脉压(mPAP)和右心室收缩末压(RVSP)均高于对照组(P均＜0.01),提示PAH模型建立成功.MCT-4w组大鼠mPAP和RVSP亦均高于对照组(P均＜0.01),但与MCT-3w组比较差异无统计学意义.对照组、MCT-2w组、MCT-3w组和MCT-4w组大鼠右心室肥厚指数呈递增趋势,MCT-3w组和MCT-4w组与对照组比较差异均有统计学意义,P分别＜0.05和0.01.(2)左、右心室心肌组织形态学:HE结果显示对照组大鼠左、有心室组织心肌细胞排列整齐,肌纤维走行一致,心肌细胞胞核清楚.MCT-3w组大鼠心肌细胞则排列紊乱,肌纤维走行不清晰,心肌细胞肌质溶解呈网状,肌间质增大,模型组各亚组间左、右心室组织形态学差异不明显.(3)左、右心室心室壁细胞中PCNA和TUNEL表达:对照组、MCT-2w组、MCT-3w组和MCT-4w组大鼠左、右心室室壁细胞数呈递增趋势,PCNA阳性细胞比例递增;而TUNEL阳性细胞比例递减,且MCT-3w组和MCT-4w组大鼠右心室室壁细胞中阳性细胞数多于左心室室壁细胞(P均＜0.05).(4)左、右心室心肌组织中糖酵解途径候选基因的筛选:MCT-4w组大鼠左心室心肌组织中HK1、HK2、PDHα1和LDHA基因表达水平均高于对照组(P均＜0.05).MCT-4w组大鼠右心室心肌组织LDHA基因表达水平高于对照组(P＜0.05),而MCT-3w组和MCT-4w组HK1基因表达水平则均高于对照组(P均＜0.05),且MCT-4w组高于MCT-3w组(尸＜0.05).(5)左、右心室心肌组织中HK1阳性细胞表达及其蛋白表达:免疫组织化学结果表明,对照组、MCT-2w组、MCT-3w组和MCT-4w组大鼠左、右心室心肌组织中HK1阳性细胞表达递增,且MCT-3w组和MCT-4w组大鼠右心室的阳性数均多于左心室(P均＜0.05).MCT-3w组和MCT-4w组大鼠左、右心室心肌组织中HK1蛋白表达水平均高于对照组(P均＜0.05).结论 PAH时左、右心室均发生能量代谢转移,右心室HK1表达早于左心室,提示干预右心室糖酵解途径可能成为延缓PAH右心衰竭的策略之一.
    목적 탐토야백합감(MCT)소치폐동맥고압(PAH)대서좌、우심실능량대사전이급가능적차이위점.방법 성년Sprague-Dawley대서,경배부피하일차성주사MCT(50 mg/kg)건립PAH우심쇠갈모형,동시설립대조조,모형조안MCT작용시간우분위3조,즉MCT-2주조、MCT-3주조화MCT-4주조(MCT-2w조、MCT-3w조화MCT-4w조).채용우심도관법검측각조대서혈액동역학지표,우심실/(좌심실+실간격)중량계산우심실비후지수,면역조직화학법검측심실벽세포증식세포핵항원(PCNA)이급원위말단전이매표기(TUNEL)적변화,실시정량PCR방법종당효해도경후선위점중사선표체차이기인.결과 (1)혈액동역학지표급우심실비후지수:MCT-3w조대서평균폐동맥압(mPAP)화우심실수축말압(RVSP)균고우대조조(P균＜0.01),제시PAH모형건립성공.MCT-4w조대서mPAP화RVSP역균고우대조조(P균＜0.01),단여MCT-3w조비교차이무통계학의의.대조조、MCT-2w조、MCT-3w조화MCT-4w조대서우심실비후지수정체증추세,MCT-3w조화MCT-4w조여대조조비교차이균유통계학의의,P분별＜0.05화0.01.(2)좌、우심실심기조직형태학:HE결과현시대조조대서좌、유심실조직심기세포배렬정제,기섬유주행일치,심기세포포핵청초.MCT-3w조대서심기세포칙배렬문란,기섬유주행불청석,심기세포기질용해정망상,기간질증대,모형조각아조간좌、우심실조직형태학차이불명현.(3)좌、우심실심실벽세포중PCNA화TUNEL표체:대조조、MCT-2w조、MCT-3w조화MCT-4w조대서좌、우심실실벽세포수정체증추세,PCNA양성세포비례체증;이TUNEL양성세포비례체감,차MCT-3w조화MCT-4w조대서우심실실벽세포중양성세포수다우좌심실실벽세포(P균＜0.05).(4)좌、우심실심기조직중당효해도경후선기인적사선:MCT-4w조대서좌심실심기조직중HK1、HK2、PDHα1화LDHA기인표체수평균고우대조조(P균＜0.05).MCT-4w조대서우심실심기조직LDHA기인표체수평고우대조조(P＜0.05),이MCT-3w조화MCT-4w조HK1기인표체수평칙균고우대조조(P균＜0.05),차MCT-4w조고우MCT-3w조(시＜0.05).(5)좌、우심실심기조직중HK1양성세포표체급기단백표체:면역조직화학결과표명,대조조、MCT-2w조、MCT-3w조화MCT-4w조대서좌、우심실심기조직중HK1양성세포표체체증,차MCT-3w조화MCT-4w조대서우심실적양성수균다우좌심실(P균＜0.05).MCT-3w조화MCT-4w조대서좌、우심실심기조직중HK1단백표체수평균고우대조조(P균＜0.05).결론 PAH시좌、우심실균발생능량대사전이,우심실HK1표체조우좌심실,제시간예우심실당효해도경가능성위연완PAH우심쇠갈적책략지일.
    Objective To explore potential divergent glycolytic metabolism gene changes between left and right ventricle in the monocmtaline (MCT) induced pulmonary arterial hypertension (PAH) rat model.Methods PAH was induced by a single subcutaneous injection of MCT (50 mg/kg) in rats.Control rats were injected with normal saline.MCT-PAH rats were randomly divided into MCT-2week,MCT-3week and MCT-4week groups (MCT-2w,3w,4w).At the end of study,the hemodynamics and right ventricular hypertrophy were compared among groups.The expression levels of proliferating cell nuclear antigen (PCNA) and TdT-mediated dUTP nick end labeling (TUNEL) in left and right ventricular cells were compared.The glycolytic key candidate genes expression was screened between two ventricles.Results After three to four weeks MCT injection,mean pulmonary arterial pressure,right ventricular systolic pressure and right ventricular hypertrophy index were all significantly increased compared to control group (all P ＜ 0.05).Both left and right ventricular morphology and structure changes were observed in all PAH rats and were similar between left and right ventricular cells.Left and right ventricular cells increased while apoptotic cells decreased in proportion to the duration post MCT injection and the PCNA positive cells in the right ventricle were higher than in the left ventricle in rats post 3 and 4 weeks MCT injection (P ＜0.05).The HK1,HK2,PDHα1 and LDHA mRNA expression in the left ventricle and LDHA mRNA expression were significantly upregulated after 4 weeks MCT injection compared to control rats (all P ＜ 0.05).Moreover,HK1 mRNA expression in the left ventricle was significantly higher in the MCT-PAH-4w group than in MCT-PAH-3w group (P ＜0.05).Immunohistochemistry analysis evidenced increasing HK1 positive cells in both left and right ventricle in proportion to MCT injection time and positive HK1 cells were significantly higher in the right ventricle than in left ventricle of MCT-PAH-3w and MCT-PAH-4w rats.Furthermore,the HK1 protein expression in left ventricular tissue form MCT-PAH-4w group and in right ventricular tissue from MCT-PAH-3w and MCT-PAH-4w groups were also significantly upregulated compared to control group (P ＜ 0.05).Conclusions Energy metabolic shift occurs both in the left and right ventricles in this PAH model.Upregulated HK1 expression appeares earlier in right ventricle compared to left ventricle.Interference on right ventricular glycolysis may be a potential novel therapy target of PAH.
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