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用免疫组织化学相邻切片多重标记技术，观察了ＣＣＫ－８与Ｍ－ＥＮＫ在小肠ＥＣ细胞中的共存，并结合图像分析技术研究同一细胞中共存物质含量的关系。实验取猪十二指肠（ｎ＝５），Ｂｏｕｉｎ液固定，制备１μｍ厚连续石蜡切片。在相邻切片上分别用兔抗５－ＨＴ、ＣＣＫ－８和Ｍ－ＥＮＫ抗体进行ＰＡＰ法免疫组织化学染色。结果见５－ＨＴ、ＣＣＫ－８和Ｍ－ＥＮＫ免疫反应细胞的形态、分布相似，主要分布于肠腺的基底部。比较观察所见，大部分５－ＨＴ阳性细胞也同时显示ＣＣＫ－８和Ｍ－ＥＮＫ阳性。ＣＣＫ－８与Ｍ－ＥＮＫ免疫染色强度有共深或共浅的倾向。将所测得的各细胞５－ＨＴ、ＣＣＫ－８和Ｍ－ＥＮＫ免疫染色灰度进行相关与回归分析，显示ＣＣＫ－８与Ｍ－ＥＮＫ免疫反应强度呈正线性相关（ｎ＝３５，ｒ＝０．５６，Ｐ＜０．０１）。而ＣＣＫ－８与５－ＨＴ、Ｍ－ＥＮＫ与５－ＨＴ的免疫反应强度之间未显示出明显的相关性。本研究证明ＣＣＫ－８和Ｍ－ＥＮＫ共存于猪十二指肠ＥＣ细胞中。同一细胞中这两种神经肽在含量上的相关性提示其在合成、释放和功能上可能相互影响。
The immunohistochemical adjacent sections multiplex labeling technique was used to observe the coexistence of CCK-8 and M-ENK in EC cells of small intestine. The relationship between CCK-8 and M-ENK in EC cells of small intestine was also studied. Pig duodenum (n = 5) was taken experimentally and fixed with Bouin solution to prepare 1μm thick continuous paraffin sections. Rabbit anti-5-HT, CCK-8 and M-ENK antibodies were respectively used for PAP immunohistochemical staining on adjacent sections. The results showed that the morphology of 5-HT, CCK-8 and M-ENK immunoreactive cells were similar and distributed mainly in the basal part of the glandular gland. In comparison, most of the 5-HT positive cells also showed CCK-8 and M-ENK positive. The intensity of CCK-8 and M-ENK immunostaining tended to be co-deep or shallow. Correlation and regression analysis of the measured immunostaining intensity of 5-HT, CCK-8 and M-ENK showed that there was a positive linear correlation between CCK-8 and M-ENK immunoreactivity (n = 35, r = 0 .56, P <0.01). There was no significant correlation between the immunoreactivity of CCK-8 and 5-HT, M-ENK and 5-HT. This study demonstrates that CCK-8 and M-ENK coexist in porcine duodenal EC cells. The correlation between the content of these two neuropeptides in the same cell suggests that they may interact with each other in their synthesis, release and function.