本文選題：C57BL/6J小鼠　切入點：心肌梗死模型　出處：《南京醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：建立可靠、穩(wěn)定的C57BL/6J小鼠缺血性心肌梗死模型,為研究藥物治療心肌梗死的機制奠定基礎。 方法：C57BL/6J小鼠經(jīng)4-6小時禁食禁飲,以戊巴比妥鈉麻醉后,仰臥固定于手術(shù)臺,經(jīng)口氣管插管,連接呼吸機,逐層分離胸部皮膚、皮下組織及肌肉,暴露心臟,于左心耳下方2～3毫米處結(jié)扎冠狀動脈。手術(shù)成功標準包括(1)結(jié)扎后心肌肉眼觀局部蒼白,心電圖示ST段抬高；(2)術(shù)后2天內(nèi)心臟超聲示小鼠左室射血分數(shù)下降并小于50%。 結(jié)果：所有的C57BL/6J小鼠經(jīng)結(jié)扎手術(shù)均被成功制作成心肌梗死模型。所有C57BL/6J小鼠心肌梗死后3周內(nèi)都發(fā)生心肌重構(gòu),心臟超聲示小鼠左室射血分數(shù)等指標進行性下降,心腔擴大,室壁變薄,部分小鼠心肌梗死后死亡。 結(jié)論：經(jīng)開胸冠狀動脈結(jié)扎手術(shù)是一種穩(wěn)定、可靠并且能夠廣泛應用的制作缺血性心肌梗死模型的手段。用小鼠心肌梗死模型來研究藥物對心肌梗死后的治療機制是一種簡單、可靠、方便、實用的方法。 目的：探討五味子乙素是否能夠改善心梗小鼠心功能及其機制的研究。 方法：C57BL/6J小鼠經(jīng)冠狀動脈結(jié)扎術(shù)后分為三組：假手術(shù)組(n=10)；心梗未治療組(n=20)；五味子乙素治療組(n=20,五味子乙素80mg/kg/day灌胃),并且在手術(shù)后持續(xù)給予3周。手術(shù)后三周觀察各組小鼠生存率,通過二維超聲心動圖檢測心功能各項指標,處死小鼠計算心臟體重比、肺臟體重比,通過Evans藍和TTC染色計算心梗面積,制作心臟病理切片進行HE染色以及Masson染色。通過免疫熒光方法檢測缺血組織中TUNEL、BrdU的表達。通過ELISA方法檢測]TNF-α、TGF-β1、IL-1β的表達,Western Blot方法檢測NF-κB,Bax,Bcl-2, eNOS,phospho-eNOS,GATA4蛋白的表達。同時培育H9c2心肌細胞,給予五味子乙素預處理后置于缺血缺氧環(huán)境,驗證該藥物在細胞水平上的作用,以及這些作用的藥物劑量依賴性。 結(jié)果：相比心梗未治療組,五味子乙素治療組心梗小鼠的死亡率在明顯降低。五味子乙素治療能夠改善心梗小鼠的心功能,提高LVEF,LVFS等心功能指標,并對假手術(shù)小鼠的心功能不產(chǎn)生影響。五味子乙素治療能夠減少心梗小鼠的心臟梗死面積,降低TNF-α、TGF-β1、IL-1β、NF-κB等炎性因子的表達,而且能夠明顯地活化內(nèi)皮一氧化氮合成酶,通過上調(diào)Bcl-2/Bax抑制缺血心肌細胞的凋亡,并且具有一定的心肌修復能力。 結(jié)論：心肌梗死對小鼠的缺血心肌細胞產(chǎn)生炎癥、纖維化、凋亡的作用。五味子乙素治療能夠明顯改善心梗小鼠的心臟功能,延緩心肌重構(gòu),減少心臟梗死面積,改善缺血心肌細胞的炎癥、纖維化、凋亡作用,加強缺血心肌細胞的修復,從而提高心梗小鼠的生存率。
[Abstract]:Aim: to establish a reliable and stable model of ischemic myocardial infarction in C57BL/6J mice. Methods after 4-6 hours fasting, the mice were anesthetized with pentobarbital sodium and fixed on the operating table. The mice were intubated with oral tube and connected to the ventilator. The chest skin, subcutaneous tissue and muscle were separated layer by layer to expose the heart. Coronary artery was ligated at 2mm below the left atrial appendage. The successful criteria included 1) local paleness of cardiac muscle eye after ligation, and ECG showed St segment elevation 2) the left ventricular ejection fraction was decreased and less than 50% by echocardiography within 2 days after operation. Results: all C57BL/6J mice were successfully made into myocardial infarction model by ligation. Myocardial remodeling occurred in all C57BL/6J mice within 3 weeks after myocardial infarction. The left ventricular ejection fraction was decreased and the heart cavity was enlarged by echocardiography. The wall became thinner and some mice died after myocardial infarction. Conclusion: transthoracic coronary artery ligation is a stable procedure. It is a simple, reliable, convenient and practical method to study the mechanism of treatment after myocardial infarction by using mouse myocardial infarction model. Aim: to investigate whether Schisandrin B can improve cardiac function and its mechanism in myocardial infarction mice. Methods: C57BL / 6J mice were divided into three groups after coronary artery ligation: sham operation group (n = 10), myocardial infarction group (n = 20), Schisandrin B group (n = 20) and Schisandrin B (80mg/kg/day) administered intragastrically for 3 weeks. Survival rate of mice in each group, The indexes of cardiac function were measured by two-dimensional echocardiography. The heart weight ratio, lung weight ratio and myocardial infarction area were calculated by Evans blue and TTC staining. The expression of Tunel BrdU in ischemic tissue was detected by immunofluorescence method. The expression of TNF- 偽 TGF- 尾 1 尾 IL-1 尾 was detected by ELISA method. The expression of NF- 魏 B Baxon Bcl-2, eNOSphospho-eNOSGATA4 protein was detected by Western Blot. Schisandrin was pretreated with Schisandrin in ischemic and hypoxic environment to verify the effect of Schisandrin on cell level and its dose-dependent effects. Results: compared with untreated myocardial infarction group, the mortality of myocardial infarction mice in Schisandra B treatment group was significantly decreased, and Schisandrin B treatment could improve cardiac function and increase LVEFV LVFS in myocardial infarction mice. Schisandra B can reduce the infarct size of heart, decrease the expression of inflammatory factors such as TNF- 偽, TGF- 尾 1, IL-1 尾 and NF- 魏 B, and activate endothelial nitric oxide synthase. Up-regulation of Bcl-2/Bax inhibits apoptosis of ischemic cardiomyocytes and has certain ability of myocardial repair. Conclusion: myocardial infarction can induce inflammation, fibrosis and apoptosis in ischemic cardiomyocytes of mice. Schisandra B can obviously improve heart function, delay myocardial remodeling and reduce infarct size in myocardial infarction mice. Improve the inflammation, fibrosis, apoptosis of ischemic cardiomyocytes, strengthen the repair of ischemic cardiomyocytes, thus improve the survival rate of myocardial infarction mice.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R285.5

【參考文獻】

相關期刊論文 前1條

1 齊治,崔景榮,田建柱,秦波,樓之岑;五味子果實揮發(fā)油對中樞神經(jīng)系統(tǒng)的藥理作用研究[J];北京醫(yī)科大學學報;1988年06期

，

本文編號：1652729