【作者】 王从军；

【导师】 刘亚黎；

【作者基本信息】 华中科技大学 ， 儿科学， 2007， 博士

【摘要】 本实验共分三部分,分别从细胞、动物模型及临床试验三方面研究柔红霉素对心肌细胞脑利钠肽表达的影响。第一部分柔红霉素对大鼠心肌细胞脑利钠肽表达的影响目的:研究柔红霉素(daunorubicin, DNR)对大鼠心肌细胞脑利钠肽(brain natriuretic peptide, BNP)表达的影响及1、6-二磷酸果糖的干预效果,进一步探讨DNR对心肌细胞BNP分泌的作用。方法:体外分离培养心肌细胞,实验分正常对照组、DNR组(DNR 0.2mg/L加入培养液)及DNR+1、6-二磷酸果糖(fructose-1,6-diphosphate FDP)组(DNR0.2mg/L及FDP2mg/ml分别加入培养液),用MTT、RT-PCR及WESTERN BLOT等方法检测各组细胞增殖及BNP表达。结果:应用柔红霉素后心肌细胞活性降低,BNP表达受抑制合成减少,加入1、6-二磷酸果糖干预后其表达并无明显增加。结论:柔红霉素对心肌细胞BNP表达呈抑制作用,果糖二磷酸不能逆转其抑制作用。第二部分柔红霉素对大鼠心肌细胞脑利钠肽含量影响的研究目的:观察柔红霉素对大鼠心肌细胞脑利钠肽(brain natriu -retic peptide, BNP)含量的影响,探讨BNP对早期诊断柔红霉素心肌病的价值。方法:32只wistar大鼠随机分成两组(每组16只):对照组(生理盐水1.0ml/kg,每3天1次×6次)。实验组(DNR 2.0mg/ kg,每3天1次×6次)。给药前后分别用比色法测定大鼠血清心肌酶谱(LDH、CK)及心肌匀浆丙二醛(malondial -dehyde,MDA)的含量。采用S-P免疫组织化学及WESTERN-BLOT方法测定心肌组织中BNP的含量。结果:给药前后对照组大鼠心肌酶谱、MDA及心肌组织BNP含量均无明显改变(P >0.05),而实验组在给药后MDA、BNP含量较给药前显著增加(P <0.001),同时实验组大鼠在应用DNR后其心肌组织BNP含量较对照组应用生理盐水后BNP含量高(P <0.001)。两组心肌酶谱在给药前后均无显著性改变(P >0.05)。结论:DNR对心肌的损害与自由基作用有关。BNP是一个较心肌酶谱更为敏感的指标,可试用于对柔红霉素心肌病的监测。第三部分急性白血病患儿使用柔红霉素后其血浆脑钠素浓度改变及其临床意义的研究目的:通过观察柔红霉素(daunorubicin DNR)化疗后急性白血病患儿血浆脑钠素(brain natriuretic peptide BNP)浓度及心脏功能指标的改变,探讨脑钠素浓度变化在早期评价柔红霉素对心脏功能影响方面的临床意义。方法:选择本院2002-2004年新诊断的急性淋巴细胞性白血病患儿并应用柔红霉素+长春新碱+左旋门冬酰胺酶+泼尼松(DVLP)方案诱导化疗的病例31例,在应用柔红霉素前后用酶联免疫吸附试验(enzyme linked immunosorbent assay ELISA)测定血浆BNP浓度,同时常规在化疗前后检测心电图(electrocardiography ECG)及心肌酶谱(LDH1、CPK-MB)。结果:应用DNR化疗后血浆BNP浓度从(3.97±2.41/)ng/L增加到(18.25±7.63)ng/L(p<0.001)。而EKG、心肌酶谱在治疗前后均无显著性改变。结论:血浆BNP浓度可作为早期监测DNR对心脏功能影响的一个有用指标。更多还原

【Abstract】 This studies were composed of three parts and were desined to investgate the effects of DNR on the expression of Brain Natriuretic Peptide in cultured rat myocardial cell、animal model and clinical tests.Part 1 Effects of DNR on the Expression of Brain Natriuretic Peptide Gene in Cultured Rat Myocardial CellsObjective:To investigate the effect of daunorubicin(DNR) on the gene expression of brain natriuretic peptide (BNP) in rat myocardial cells.Methods:Neona -tal WISTAR rat myocardial cells were cultured and were divided into three groups,including control group、DNR group(DNR 0.2mg/L) and DNR+ fructose-1, 6- diphosphate (FDP )group(DNR 0.2mg/L、FDP2mg/ml).The viability of myocardial cells and the expression of BNP gene were examined by MTT、RT-PCR and WESTERN BLOT.Results:The Viability of myocardial cells precipitous decreased with the increasing of concentrations of DNR. The expression and the synthesis of BNP decreased after receving DNR and no increasing were observed after receving FDP to inter-vene.Conclusions:DNR suppressed the expression of BNP gene in cultured rat myocardial cells and FDP couldn’t reverse the suppressing effect.Part 2 Investigation to the Brain Natriuretic Peptide Content of Myocardium in Rats Induced by DaunorubicinObjective:To explore the value of brain natriuretic peptide (BNP) in diagnosing the myocardial induced by daunorubicin(DNR) by investigate the changes of the BNP level in myocardium tissue of the rat after administrating the DNR. Methods:Thirty-two male young wistar rats were randomly divided into two groups(n=16 in each).The first group served as control group(0.9%saline 1.0ml/kg q.3d×6).The other group received DNR (2.0mg/kg q.3d×6) by tail vein injection. The content of serum LDH、CK and myocardium tissue malondialdehyde (MDA) were measured by colorimetric method and at the time the level of myocardium tissue BNP were analyzed by s-p immunohistochemistry and WESTERN-BLOT after administrating the drug.Results:The levels of MDA and BNP in myocardium tissue increase (P <0.001)and no significant change were observed in the serum LDH and CK(P >0.05) after administrating DNR in experiment group. On the other hand there is no significant change in control group. Furthermore,the levels of BNP was higher in experiment group than in control group after receving DNR and saline respectively.Conclusions:BNP is a good indicator of DNR-induced myocardial.Part 3The changes and the clinical values of plasma brain natriuretic peptide concentration in children with acute leukemia accepted the chemotherapy of daunorubicinObjective:This study was performed to explore the clinical values of the brain natriuretic peptide(BNP) in evaluating the cardiotoxicity caused by daunorubicin( DNR )by studying the changes of the plasma BNP level of children with acute leukemia that accepted the chemotherapy with DNR.Methods:31 children with acute lymphoblastic leukemia were performed the induction chemotherapy in DVLP project. We measured the plasma level of BNP by enzyme linked immunosorbent assay( ELISA) at the beginning and completing of administrating DNR. Simultaneously, electrocardiography (ECG )and cardiac muscle enzymes(LDH1、CPK-MB) were measured as a rule.Results:The plasma level of BNP increased from (3.97±2.41/)ng/L to(18.25±7.63)ng/L(p <0.001). However, routing EKG and cardiac muscle enzymes(LDH1 CPK-MB) remained normal.Conclusions:The plasma level of BNP was more sensitive in early evaluating the heart function during the chemotherapy of DNR.更多还原