论文部分内容阅读
本实验通过观察一氧化氮合酶(nNOS)与细胞内钙结合蛋白(calbindin, CB)共存情况,旨在研究大鼠食管内NO类神经元的神经化学特性并试图寻找出引起食管NO类神经元在衰老过程中丢失的可能原因。取自不同年龄组及种系(Wistar和S-D)大鼠的食管组织,制成肌间神经丛铺片,经nNOS与CB免疫组化双重染色后在荧光显微镜下观察。结果显示,nNOS与CB的免疫阳性反应物均见于大鼠食管肌间神经元胞体内。根据二者共存情况,大鼠食管内nNOS免疫阳性神经元可分为两大亚类:nNOS+/CB+及nNOS+/CB-。而且我们首次发现在大鼠食管腹腔段含有大量nNOS+/CB+神经元,约占NO类神经元总体的30% ~40%,与食管其它部位相比有显著差异(P<0.001)。另外,大鼠食管腹腔段绝大多数CB免疫阳性神经元同时也为nNOS免疫阳性(约占95% ~100%)。上述特征均见于本实验所观察的所有不同年龄及不同种系的大鼠食管内。在大鼠衰老过程中,nNOS+/CB+神经元的相对百分比的变化具有种系特异性。线形回归实验分析表明:年轻SD大鼠食管内nNOS+/CB+神经元的百分比含量与衰老过程中NO类神经元丢失数量的百分比之间存在显著负相关性(correlation coefficient 0.99;P<0.05)。结果提示在S-D大鼠衰老过程中其食管内NO类神经元的丢失可能与其细胞内钙结合蛋白含量的变化有关。但nNOS/CB共存神经元在分布上有明显差异的生物学意义尚有待于进一步研究。
In this study, we observed the coexistence of nitric oxide synthase (nNOS) and calbindin (CB) in order to study the neurochemical characteristics of NO neurons in the esophagus and try to find out the neurotrophic factor Possible causes of the loss of meta-elements during aging. Esophageal tissues from different age groups and germline (Wistar and S-D) rats were made into myenteric plexuses and were observed under a fluorescence microscope after double staining with nNOS and CB immunohistochemistry. The results showed that nNOS and CB immunoreactive reactants are found in the rat esophageal myenteric neurons. According to the coexistence of the two, nNOS immunoreactive neurons in the rat esophagus can be divided into two major categories: nNOS + / CB + and nNOS + / CB-. Moreover, we found for the first time that a large amount of nNOS + / CB + neurons were found in the peritoneal cavity of the esophagus, accounting for about 30-40% of the total number of NO neurons, which was significantly different from other parts of the esophagus (P <0.001). In addition, the vast majority of CB immunoreactive neurons in the esophageal peritoneal cavity of rats are also immunologically positive for nNOS (about 95% to 100%). The above characteristics are found in the experiment observed in all different ages and different strains of rat esophagus. The relative percentage change in nNOS + / CB + neurons is germ-line specific during rat senescence. The linear regression analysis showed that there was a significant negative correlation between the percentages of nNOS + / CB + neurons in the esophagus and the percentages of NO-like neurons in aging rats (correlation coefficient 0.99; P <0.05). The results suggest that the loss of NO neurons in the esophagus may be related to the change of intracellular calcium-binding protein content in S-D rats during aging. However, the biological significance of the significant differences in the distribution of nNOS / CB co-existing neurons remains to be further studied.