Clinic of Small Animal Medicine, LMU University of Munich, Veterinaerstrasse 13, Munich 80539, Germany.
BMC Vet Res. 2013 Jan 17;9:12. doi: 10.1186/1746-6148-9-12.
Dilated cardiomyopathy (DCM) is the most common heart disease in Doberman Pinschers. MicroRNAs (miRNAs) are short non-coding RNAs playing important roles in gene regulation. Different miRNA expression patterns have been described for DCM in humans and might represent potential diagnostic markers. There are no studies investigating miRNA expression profiles in canine DCM. The aims of this study were to screen the miRNA expression profile of canine serum using miRNA microarray and to compare expression patterns of a group of Doberman Pinschers with DCM and healthy controls.
Eight Doberman Pinschers were examined by echocardiography and 24-hour-ECG and classified as healthy (n=4) or suffering from DCM (n=4). Total RNA was extracted from serum and hybridized on a custom-designed 8x60k miRNA microarray (Agilent) containing probes for 1368 individual miRNAs. Although total RNA concentrations were very low in serum samples, 404 different miRNAs were detectable with sufficient signal intensity on miRNA microarray. 22 miRNAs were differentially expressed in the two groups (p<0.05 and fold change (FC)>1.5), but did not reach statistical significance after multiple testing correction (false discovery rate adjusted p>0.05). Five miRNAs were selected for further analysis using quantitative Real-Time RT-PCR (qPCR) assays. No significant differences were found using specific miRNA qPCR assays (p>0.05).
Numerous miRNAs can be detected in canine serum. Between healthy and DCM dogs, miRNA expression changes could be detected, but the results did not reach statistical significance most probably due to the small group size. miRNAs are potential new circulating biomarkers in veterinary medicine and should be investigated in larger patient groups and additional canine diseases.
扩张型心肌病(DCM)是杜宾犬中最常见的心脏病。microRNAs(miRNAs)是短的非编码 RNA,在基因调控中发挥重要作用。人类 DCM 已有不同的 miRNA 表达模式,并可能代表潜在的诊断标志物。目前尚无研究调查犬 DCM 的 miRNA 表达谱。本研究旨在使用 miRNA 微阵列筛选犬血清中的 miRNA 表达谱,并比较一组患有 DCM 和健康对照的杜宾犬的表达模式。
通过超声心动图和 24 小时心电图检查了 8 只杜宾犬,并将其分为健康(n=4)或患有 DCM(n=4)。从血清中提取总 RNA,并在定制的 8x60k miRNA 微阵列(Agilent)上杂交,该微阵列包含 1368 个单个 miRNA 的探针。尽管血清样本中的总 RNA 浓度非常低,但 miRNA 微阵列上仍可检测到 404 种不同的 miRNA,且信号强度足够。两组间有 22 个 miRNA 表达差异(p<0.05 和倍数变化(FC)>1.5),但经过多次测试校正(调整后的错误发现率 p>0.05)后未达到统计学意义。选择了 5 个 miRNA 进行定量实时 RT-PCR(qPCR)检测。使用特定的 miRNA qPCR 检测未发现显著差异(p>0.05)。
犬血清中可检测到大量的 miRNA。在健康和 DCM 犬之间,可以检测到 miRNA 表达的变化,但由于样本量小,结果未达到统计学意义。miRNAs 是兽医领域有潜力的新型循环生物标志物,应在更大的患者群体和其他犬类疾病中进行研究。