| ObjectiveTo study the structure of different acupoints located in the skin using traditional histological and immunohistochemical method. The locations and quantities of blood vessels , nerves , fibers and receptors in particular points and non-points are investigated firstly, and the Cx43 expressions of both points and non-points with and without stimulation of needle in the morphological structures of points are partially revealed, are investigated .Materials and methodsTwenty male Wistar rats were selected and divided into two groups randomly: normal control group and group with stimulation of acupoint, in which each has 10 rats. Rats in normal control group were killed directly through air embolism, cleared and fixed with 10% formaldehyde solution for 15 days. For rats in group with stimulation, particular acupoints including "Nieguan" point at upper limbs, "Zusanli" point at lower limbs, "Dazhui" point at backside, "Guanyuan" point at abdominal zone and "Yongquan" at claws were selected to be stimulated by needle。 The stimulating were then given by once per 10 minutes,the frequency was with 1-2 times /sec, and the rats were killed through air embolism 30 minutes after stimulation, the following treated method is same as that in normal control group. Extract 0.5×0.5×0.5cm block of tissue at the point and 1cm away from the point of rats in normal control group, and extract the tissue block of same size at point of rats in group with stimulation. Because the area of bottom of root where "Yongquan" point located at is small, no skin for normal control group was extracted. The extracted tissue was cut into frozen sections with thickness of 30μm. The sections were stained by HE method , Masson 3-Color method, improved Bielschowsky method and immunohistochemical method (CX43) separately. Labeled cells were observed withlight microscope. Statistical analysis between the two groups was performed with one-wayANOVA and q test.Results1. Contrast between points and non-points.In "Zusanli" point group, there were significant differences in quantities of blood vessel, nerves and collagenic fibers between points and non-points. In "Guanyuan" point group, there was significant difference in quantity of nerve branches between points and non-points, while there is no significant difference in quantities of blood vessels and collagenic fibers. In "Neiguan" and "Dazhui" point groups, there was no significant difference between points and non-points. Due to the difficulty of tissue extracting, there is no data for "Yongquan" point.2. Contrast between different pointsAs shown in table 2, the difference varied for all the studied points. There was no significant difference in quantity of branch veins between "Neiguan" and "Zusanli" point groups;While either "Guanyuan" or "Dazhui"point groups show an obviously decrease of vein branches contrasted to "Zusanli" group. There was a significant decrease of branch nerves at "Neiguan","Guanyuan" or "Dazhui" point than "Zusanli" point , while a significant increase at "Yongquan" point. "Neiguan","Guanyuan" or "Dazhui" point has fewer nerve branches than "Zusanli" point, while "Yongquan" point has more nerve branches than "Zusanli". There was no significant difference in quantity of collagenic fibers between "Neiguan", "Guanyuan", "Dazhui" and "Zusanli" point groups, while there was a much rich expression of collagenic fibers at "Yongquan" than the others.In addition, as shown in stained sectors, the type and quantity of receptors is different at studied point groups. At skin level, there were mainly dissociative nerve endings at "Zusanli" and "Neiguan" point, tactile corpuscles and meissner's corpuscles are occasionally observed;Af'Guanyuan" and "Dazhui" point areas, thereare mainly receptors of hair flooicle and dissociative nerve endings;At "Yongquan" point area, pacinian corpuscles, tactile corpuscles and dissociative nerve endings are of large quantity, Ruffini body are only occasionally observed. Because Krause are usually located at conjunctiva or mucous membrane, they are not observed at skin level of those studied points. Due to the limited filed of vision, there are few receptors sometimes even none can be observed. The statistic result of receptors is not very precise, and needs to be improved in the future research work.3. Contrast of Cx43 Expression between points and non-points, before and after stimulationAs shown in the immunohistochemical stain result, Cx43 expression was mainly in cytoplasm and cytolemma of epithelial cells including formative cells that form epidermis and hair flooicles, especially in basal lamina of epidermis and prickle-cell layer, and is less in homy layer. There were significant differences in Cx43 expression between point groups with stimulation and those without stimulation;Cx43 expression at "Zusanli" point in group without stimulation was significantly higher than at non-points in control group, and there was a very significant difference between these two groups;Cx43 expression at "Zusanli" point after stimulation increased significantly, and it was a significantly difference to that at non-points in normal control groups. Cx43 expression increases at points in groups with stimulation contrast to those without stimulation;Cx43 expression increases partially at non-points after stimulation , however, it was not a significant difference to that at non-points without stimulation. In "Neiguan", "Guanyuan" and "Dazhui" point groups, the results were same as those in "Zusanli" point groups. In "Yongquan" point group, Cx43 expression increases significantly after stimulation contrasted to group without stimulation.ConclusionThere is no significant statistically difference in anatomic structure, except forparticular points. However, from view of molecular biology, as shown in results of Connexin expression through immunohistochemical staining method, there are significant differences between points and non-points, which indicates that the points are actually existed and have special characteristics. |
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