Ikeda Ayami, Kai Hisashi, Kajimoto Hidemi, Yasuoka Suguru, Kage Masayoshi, Imaizumi Tsutomu
Division of Cardio-Vascular Medicine, Department of Internal Medicine, Kurume University School of Medicine, 67 Asahimachi, Kurume, Fukuoka 830-0011, Japan.
Int J Vasc Med. 2012;2012:863410. doi: 10.1155/2012/863410. Epub 2012 Jun 12.
Background. The heart consists of various kinds of cell components. However, it has not been feasible to separately analyze the gene expression of individual components. The laser microdissection (LMD) method, a new technology to collect target cells from the microscopic regions, has been used for malignancies. We sought to establish a method to selectively collect the muscular and vascular regions from the heart sections and to compare the marker gene expressions with this method. Methods and Results. Frozen left ventricle sections were obtained from Wistar-Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHR-SP) at 24 weeks of age. Using the LMD method, the muscular and vascular regions were selectively collected under microscopic guidance. Real-time RT-PCR analysis showed that brain-type natriuretic peptide (BNP), a marker of cardiac myocytes, was expressed in the muscular samples, but not in the vascular samples, whereas α-smooth muscle actin, a marker of smooth muscle cells, was detected only in the vascular samples. Moreover, SHR-SP had significantly greater BNP upregulation than WKY (P < 0.05) in the muscular samples. Conclusions. The LMD method enabled us to separately collect the muscular and vascular samples from myocardial sections and to selectively evaluate mRNA expressions of the individual tissue component.
背景。心脏由多种细胞成分组成。然而，单独分析各个成分的基因表达一直不可行。激光显微切割（LMD）方法是一种从微观区域收集目标细胞的新技术，已用于恶性肿瘤研究。我们试图建立一种从心脏切片中选择性收集肌肉和血管区域的方法，并比较该方法中标记基因的表达情况。方法与结果。从24周龄的Wistar-Kyoto大鼠（WKY）和易中风自发性高血压大鼠（SHR-SP）获取冷冻左心室切片。使用LMD方法，在显微镜引导下选择性收集肌肉和血管区域。实时逆转录聚合酶链反应（RT-PCR）分析表明，心肌细胞标记物脑钠肽（BNP）在肌肉样本中表达，但在血管样本中不表达，而平滑肌细胞标记物α-平滑肌肌动蛋白仅在血管样本中检测到。此外，在肌肉样本中，SHR-SP的BNP上调明显高于WKY（P < 0.05）。结论。LMD方法使我们能够从心肌切片中分别收集肌肉和血管样本，并选择性评估各个组织成分的mRNA表达。
