Takemura G, Fujiwara H, Mukoyama M, Saito Y, Matsuda M, Kawamura A, Ishida M, Kida M, Uegaito T, Nakao K
Department of Internal Medicine, Faculty of Mediciine, Kyoto University, Japan.
Jpn Circ J. 1989 Jul;53(7):686-94. doi: 10.1253/jcj.53.686.
The presence of atrial natriuretic polypeptide (ANP) was immunohistochemically demonstrated in the formalin-fixed and paraffin-embedded tissues of 25 autopsied normal human hearts using monoclonal antibody. The ANP amounts were immunohistochemically semiquantified and compared with amounts measured by radioimmunoassay (RIA). The 25 autopsied hearts were divided into 5 groups according to the interval of formalin fixation or the length of time between death and fixation. Formalin-fixation intervals were one week in group 1A and 1B; 1 year in group 2; 4 to 5 years in group 3 and 10 to 12 years in group 4. The hearts of group 1A, 2, 3 and 4 were fixed within 5 hours after death. Those of group 1B were fixed 14 to 18 hours at 4 degrees C. After fixation, the left and right atrial appendages (LAA and RAA), the left and right atrial free walls (LA and RA), the left and right ventricular free walls (LV and RV) and the ventricular septum (VS) were transmurally dissected from each heart. They were embedded in paraffin, cut into 4 microns sections and immunohistochemically stained by the avidin-biotin-peroxidase complex (ABC) method using monoclonal antibody against alpha-human ANP. Under a light microscope, they were evaluated semiquantitatively according to the incidence of ANP-positive cells and the intensity of immunostaining. For every heart in group 1A, the tissue concentrations of ANP in the different parts were also measured separately by RIA before fixation. ANP-positive myocytes were noted in the atria of all hearts of all groups, but no in any ventricular myocytes. Both their incidence and grade in the atria were similar among groups 1A, 1B and 2. However, they were less in group 3, and least in group 4 among all groups. For all groups, they decreased in the following order: LAA greater than RAA not equal to LA greater than RA; the inner 1/3 greater than the middle 1/3 greater than the outer 1/3 of the atrial walls. The order in LAA, RAA, LA and RA.(ABSTRACT TRUNCATED AT 250 WORDS)
使用单克隆抗体，通过免疫组织化学方法在25例正常人体尸检心脏的福尔马林固定石蜡包埋组织中证实了心钠素（ANP）的存在。对ANP含量进行免疫组织化学半定量，并与放射免疫测定（RIA）测得的含量进行比较。根据福尔马林固定间隔或死亡与固定之间的时间长度，将25例尸检心脏分为5组。第1A组和1B组的福尔马林固定间隔为1周；第2组为1年；第3组为4至5年；第4组为10至12年。第1A组、第2组、第3组和第4组的心脏在死亡后5小时内固定。第1B组的心脏在4℃下固定14至18小时。固定后，从每个心脏中全层切下左、右心耳（LAA和RAA）、左、右心房游离壁（LA和RA）、左、右心室游离壁（LV和RV）以及室间隔（VS）。将它们包埋在石蜡中，切成4微米厚的切片，并用抗α-人ANP单克隆抗体通过抗生物素蛋白-生物素-过氧化物酶复合物（ABC）法进行免疫组织化学染色。在光学显微镜下，根据ANP阳性细胞的发生率和免疫染色强度进行半定量评估。对于第1A组的每颗心脏，在固定前还通过RIA分别测量不同部位的ANP组织浓度。在所有组的所有心脏的心房中均发现了ANP阳性心肌细胞，但在任何心室心肌细胞中均未发现。第1A组、第1B组和第2组心房中的发生率和分级相似。然而，在所有组中，第3组较少，第4组最少。对于所有组，它们按以下顺序降低：LAA大于RAA不等于LA大于RA；心房壁的内1/3大于中1/3大于外1/3。LAA、RAA、LA和RA中的顺序。（摘要截断于250字）
