【作者】 刘科；

【导师】 黄克力；

【作者基本信息】 电子科技大学 ， 生物医学工程， 2017， 硕士

【摘要】 本次实验的目的是为了比较厄贝沙坦及辛伐他汀对大鼠心肌纤维化的影响作用,探索miRNA21,miRNA29b,miRNA133a在心肌纤维化中的表达调控及血清中的含量是否与组织表达程一致性。实验选用了体重200~220gSD雄性大鼠50只,行腹主动脉缩窄术后二周随机分为5组,灌胃给药,予假手术组及模型组0.5%羧甲基纤维素钠(0.5ml/d)、厄贝沙坦组(厄贝沙坦20mg.Kg/d)、辛伐他汀组(辛伐他汀5mg.Kg/d)及联合用药组(厄贝沙坦20mg.Kg/d+辛伐他汀5mg.Kg/d),连续8周后测定体重、血压、心脏指数、血清Ang II,超声测左室舒张末期(LVD)、左室收缩末期(LVS)、室间隔(IVS)直径,HE染色观察心肌细胞,Masson染色观察胶原、测心肌胶原容积(CVF),免疫组化观察并半定量测转化生子因子-β1(TGF-β1)、结缔组织生长因子(CTGF)及血管紧张素受体1(ATR1),Western blot定量测TGF-β1、CTGF及ATR1蛋白含量,qPCR法检测各组心肌组织及血清中miRNA21,miRNA29b,miRNA133a的表达含量。通过实验我们得出,与假手术组比,模型组的心脏指数、室间隔、CVF、TGF-β1、CTGF均增高,体重、ATR1降低;与模型组相比,厄贝沙坦组、辛伐他汀组体重增加,联合用药组体重降低,厄贝沙坦组、辛伐他汀组心脏指数、室间隔、EF、CVF、TGF-β1、CTGF、ATR1降低(P<0.05),而联合用药组除ATR1增高(P<0.05)外,其余均低于单一用药组(P<0.05)。组织中miRNA21:与假手术组相比,厄贝沙坦组降低,模型组轻微降低,辛伐他汀组、联合用药组增高;miRNA29b:与假手术组相比,辛伐他汀组增高,厄贝沙坦组、联合用药组、模型组降低;miRNA133a:与假手术组相比,厄贝沙坦组、辛伐他汀组、联合用药组、模型组均降低;血清中除miRNA29在辛伐他汀组水平略高于假手术组外,其余组表达水平均低于假手术组。因此我们认为厄贝沙坦及辛伐他汀均能缓解心肌纤维化,联合用药效果好于单一用药,尽管出现了不明原因的ATR1含量增加。需根据适应症使用以减少药物不良反应。miRNA21,miRNA29b,miRNA133a在组织中的表达与纤维化程度有一定关系,在血清中的表达均高于正常对照组,有一定意义。更多还原

【Abstract】 In this experiment Myocardial fibrosis model was established in rats,irbesartan and simvastatin were used to intervened,and investigating the regulation expression of miRNA21,miRNA29 b and miRNA133 a in myocardial fibrosis and whether the content of serum is consistent with the expression of tissue.We chose 50 male SD rats were randomly divided into 5 groups,two weeks after abdominal aortic coarctation,Intragastric administration was given to sham group and model group with 0.5% sodium carboxymethyl cellulose(0.5ml/d),irbesartan group(irbesartan 20 mg.Kg/d),simvastatin group(simvastatin 5mg.Kg/d)and combination group(irbesartan 20 mg.Kg/d+ 5mg.Kg/d simvastatin)for 8 weeks.Measure body weight,blood pressure,heart index,serum Ang II and CVF,LVD,LVS,IVS diameter measured by ultrasound,pathological staining with HE and Masson.Immunohistochemistry and Western blot are used to determine TGF-β1,CTGF and ATR1 protein content,qPCR method was used to detect the expression levels of miRNA21,miRNA29 b and miRNA133 a in tissue and serum in rats of each group.Through experiments,we know that compared with the sham group,the heart index,IVS,CVF,TGFβ1 and CTGF increased in model group,and the body weight and ATR1 decreased(P<0.05);Compared with the model group,body weight gain in irbesartan and simvastatin group,loss in combination group,cardiac index,EF,CVF,TGF-beta 1,CTGF and ATR1 decreased in irbesartan and simvastatin group(P<0.05),and except ATR1 increased in combination group(P<0.05),the rest were lower than the irbesartan and simvastatin group(P<0.05).miRNA21 in Ir group and Model group decreased,increased in Si group and Ir+Si group;miRNA29b increased in Si group,decreased in Ir group,Ir+Si group and Model group;miRNA133a decreased in Ir group,Si group,Ir+Si group and Model group.From this point of view,Irbesartan and simvastatin could alleviate myocardial fibrosis,and the combination of 2drugs is more effective,although the ATR1 increased with unkonw reason.Pay attention to choise according to indications to reduce adverse drug reactions.The expression of miRNA21,miRNA29 b,miRNA133a in the serum of rats with fibrosis are abnormal compared with the Sham group.更多还原