实验动物与比较医学
實驗動物與比較醫學
실험동물여비교의학
LABORATORY ANIMAL AND COMPARATIVE MEDICINE
2010年
5期
329-334
,共6页
王萧%王宁%林传权%许庆文%付云%刘金元
王蕭%王寧%林傳權%許慶文%付雲%劉金元
왕소%왕저%림전권%허경문%부운%류금원
心肌组织%心衰模型%动物%细胞凋亡%一氧化氮%Caspase-3
心肌組織%心衰模型%動物%細胞凋亡%一氧化氮%Caspase-3
심기조직%심쇠모형%동물%세포조망%일양화담%Caspase-3
Myocardial tissue%Congestive heart failure model,animal%Apoptosis%Nitric oxide%Caspase-3
目的 研究导管术致超容量负荷型实验性心衰兔模型随时间推移的损伤变化及其与NO的关系. 方法 运用导管术造成兔的超容量负荷型心衰模型,然后分别于造模后24 h和造模后7 d处死动物,取心脏组织做光镜检查;NO试剂盒检测血清NO含量、通过流式细胞仪检测心肌细胞凋亡情况、Western blot法检测Caspase-3蛋白水平的表达. 结果 ①光镜观察:正常组心肌细胞无损伤表现,造模24h组及7d组均有心肌细胞损伤表现,造模7 d组损伤更为广泛且严重:②NO试剂盒检测:造模24 h组NO含量(3.33±1.30 μmol/L)和造模7 d组含量(2.81±0.77/μmol/L)高于正常组(2.09±0.22 μmol/L),且造模24 h高于造模7 d;③AnnexinV联合PI染色:正常组凋亡率较低(3.72%±1.92%),随着时间推移,凋亡细胞所占比例呈上升趋势,造模7 d(10.73%±2.26%)明显高于造模24 h(7.62%±1.70%),④Western Blot检测:正常组基本检测不到激活的Caspase-3蛋白酶,在造模后24 h和造模后7 d Caspase-3蛋白酶均有明显表达,但造模7 d有所下降. 结论 超容量负荷型心衰兔模型心肌随时间推移,凋亡率增加、心肌损伤加重;NO在本心衰模型的主要作用以抗心肌细胞凋亡效应作用为主.
目的 研究導管術緻超容量負荷型實驗性心衰兔模型隨時間推移的損傷變化及其與NO的關繫. 方法 運用導管術造成兔的超容量負荷型心衰模型,然後分彆于造模後24 h和造模後7 d處死動物,取心髒組織做光鏡檢查;NO試劑盒檢測血清NO含量、通過流式細胞儀檢測心肌細胞凋亡情況、Western blot法檢測Caspase-3蛋白水平的錶達. 結果 ①光鏡觀察:正常組心肌細胞無損傷錶現,造模24h組及7d組均有心肌細胞損傷錶現,造模7 d組損傷更為廣汎且嚴重:②NO試劑盒檢測:造模24 h組NO含量(3.33±1.30 μmol/L)和造模7 d組含量(2.81±0.77/μmol/L)高于正常組(2.09±0.22 μmol/L),且造模24 h高于造模7 d;③AnnexinV聯閤PI染色:正常組凋亡率較低(3.72%±1.92%),隨著時間推移,凋亡細胞所佔比例呈上升趨勢,造模7 d(10.73%±2.26%)明顯高于造模24 h(7.62%±1.70%),④Western Blot檢測:正常組基本檢測不到激活的Caspase-3蛋白酶,在造模後24 h和造模後7 d Caspase-3蛋白酶均有明顯錶達,但造模7 d有所下降. 結論 超容量負荷型心衰兔模型心肌隨時間推移,凋亡率增加、心肌損傷加重;NO在本心衰模型的主要作用以抗心肌細胞凋亡效應作用為主.
목적 연구도관술치초용량부하형실험성심쇠토모형수시간추이적손상변화급기여NO적관계. 방법 운용도관술조성토적초용량부하형심쇠모형,연후분별우조모후24 h화조모후7 d처사동물,취심장조직주광경검사;NO시제합검측혈청NO함량、통과류식세포의검측심기세포조망정황、Western blot법검측Caspase-3단백수평적표체. 결과 ①광경관찰:정상조심기세포무손상표현,조모24h조급7d조균유심기세포손상표현,조모7 d조손상경위엄범차엄중:②NO시제합검측:조모24 h조NO함량(3.33±1.30 μmol/L)화조모7 d조함량(2.81±0.77/μmol/L)고우정상조(2.09±0.22 μmol/L),차조모24 h고우조모7 d;③AnnexinV연합PI염색:정상조조망솔교저(3.72%±1.92%),수착시간추이,조망세포소점비례정상승추세,조모7 d(10.73%±2.26%)명현고우조모24 h(7.62%±1.70%),④Western Blot검측:정상조기본검측불도격활적Caspase-3단백매,재조모후24 h화조모후7 d Caspase-3단백매균유명현표체,단조모7 d유소하강. 결론 초용량부하형심쇠토모형심기수시간추이,조망솔증가、심기손상가중;NO재본심쇠모형적주요작용이항심기세포조망효응작용위주.
Objective Cardio-cerebrovascular disease is a high incident disease in the world,the research of which depends on the establishment of animal models.The purpose of this study was to observe the myocardial changes with time in rabbit congestive heart failure models(CHF),as well as its relationship to Nitric Oxide(NO).Method Twenty-four rabbits were randomly assigned into three groups with eight rabbits in each one:normal group,model 24h group and model 7d group The model of CHF with volume overload was set up via catheterization,and then the rabbits were executed after 24h and 7d respectively,whose myocardial tissue was used for light microscopic examination.Serum NO was measured using NO assay kit,and myocardial apoptosis was detected by flow cytometry.Caspase-3 expression was determined by Western Blot analysis.Results(1)The structures of myocardial cells in normal group were intact,and there were no defects.The myocardial injury was found in the group of CHF model after 24 h and 7d,while the injury in the 7d group was more extensive and serious.(2)The contents of serum NO increased significantly in model 24h group (3.33±1.30 μmol／L)compared with the normal group(2.09±0.22 μmol／L)(P<0.05),and decreased a little in model 7d group(2.81±0.77 μmol／L).(3) The apoptosis rate were(3.72±1.92％),(7.62±1.70％)and(10.73±2.26％)respectively,which is going up as time goes on.(4)The results of Western Blot analysis showed that there was almost no expression of the active form of Caspase-3 in the normal group.The level of Caspase-3 expression was significant in the 24h group and 7d group,while the latter decreased somewhat.Conclusion As time went on,the apoptosis rate increased and myocardial injury aggravated,and the main function of NO is anti-apoptotic in the model.
