糖心康对糖尿病大鼠应用基因芯片技术标记心肌基因CYP450 2c37mRNA表达的影响

Effect of Tangxinkang on the gene chip technique labeled myocardial gene CYP450 2c37mRNA expression in diabetic rats

目的：观察糖尿病性心肌病大鼠心肌组织中CYP450 2C37mRNA的表达，以及中药复方糖心康干预后的变化。 方法：实验于2002-05／2004-09在上海博星基因公司实验室进行。将50只雄性Wistar大鼠随机分为空白组(n=10)、模型组(n=20)、糖心康 组(n=20)。①糖尿病大鼠心肌损伤模型制备：糖心康组和模型组大鼠左F腹腔注射链脲佐菌素25mg／kg，空白组注射柠檬酸缓冲液25mg／kg，然后各组均喂以高热量饲料。②给药：从造模成功后第15周开始，糖心康组以中药糖心康(太子参、黄芪、麦冬、玉竹、丹参、枸杞、土虫等组成)5μL／g(0．79g／mL)灌胃，1次／d，共7周，空白组及模型组以等量生理盐水灌胃。③观察指标：从心肌组织中抽提mRNA，经反转录分别用Cy3，Cy5荧光标记，获得3组动物来源的cDNA探针，cDNA探针与基因表达谱芯片杂交，结果用扫描仪扫描并用软件进行统计分析，筛选出Ratio值大于2或小于0．5的差异表达数据。不同心肌组织样本用双色荧光标记叠加图和杂交信号强度散点图表示。 结果：35只大鼠进入结果分析，筛选出差异表达数据392项，双点均一显著异常表达基因124个。糖尿病心肌基因CYP450 2C37mRNA表达上调，糖心康组心肌基因CYP450 2C37mRNA表达下调。 结论：CYP450 2C37mRNA在糖尿病状态下心肌也有表达，且表达上调，而中药复方糖心康可使其下调，这也是中药复方对糖尿病性心肌损伤具有保护作用的机制之一。

AIM To observe the expression of CYP450 2C37mRNA in the myocardial tissue of rats with diabetic cardiomyopathy, and the observe the changes after the intervention of the Chinese herb of Tangxinkang. METHODS： The experiment was done in the laboratory of Shanghai Boxing Gene Company from May 2002 to September 2004. Fifty male Wistar rats were randomly divided into blank group （n=10）, model group （n=20） and Tangxinkang group （n=20）. ① Establishment of myocardial damage models of diabetic rats： The rats in the model group and Tangxinkang group were treated with intraperitoneal injection of streptozotocin （25 mg/kg）, and those in the blank group were injected with citromalic acid buffer （25 mg/kg）, and then all the rats were fed with hyper-calorie diet. ② Administration： From the 15^th week after the model establishment, rats in the Tangxinkang group were treated with gastric perfusion of 5 μL/g （0.79 g/mL） the Chinese herb of Tangxinkang （composed with heterophylly falsestarwort root, membranous milkvetch root, dwarf lilyturf tuber, fragrant solomonseal rhizome, danshen root, medlar, ground beetle, etc.）, once a day for 7 weeks, and those in the blank group and model group were treated with gastric perfusion of saline. ③ Observational indexes： The mRNA was extracted from myocardial tissues firstly, and cDNA probes were obtained from the rats in the 3 groups by Cy3 and Cy5 labelling after reverse transcription, and then the cDNA probes were hybridized with gene chip, and the results were scanned with the scanning apparatus and statistically analyzed with software. The data whose ratio was more than 2 or less than 0.5 were selected. Different myocardial samples were labeled by two kinds of fluorescents and expressed with hybridization signals. RESULTS： Finally 35 rats were involved in the analysis of results. Totally 392 different expression data and 124 abnormal remarkably expression genes were obtained. The expression of diabetic myocardial gene-CYP450 2C37mRNA was up-regulated, while the expression of rat myocardial gene-CYP450 2C37mRNA in the Tangxinkang group was down-regulated. CONCLUSION： The myocardial gene-CYP450 2C37mRNA is also expressed and up-regulated when suffering from diabetes mellitus, and Chinese herb Tangxinkang can down-regulated the expression. It is one of protective mechanisms of the Chinese herb on diabetic cardiomyopathy.