急性肠黏膜损伤大鼠腹部伊文氏蓝渗出点与穴位分布特征的多元统计分析

Multivariate statistical analysis of Evans blue exudation spots and acupoint sensitization distribution in rats with acute intestinal mucosal injury

目的:结合经纬网格计数法与多元统计分析探讨急性肠黏膜损伤(AIMI)大鼠腹部伊文氏蓝(EB)渗出点的体表分布特征,并探讨该方法的可行性。方法:24只SD大鼠随机分为空白组、7.5%芥子油组、10%芥子油组、12.5%芥子油组,每组6只。采用芥子油灌肠法建立大鼠模型。大鼠尾静脉注射EB(0.1 mL/100 g),分别于注射后5、10、15、20、25 h观察并拍摄记录大鼠腹部体表EB渗出点。采用HE染色法观察结肠黏膜组织病理。构建经纬网格,记录网格位置及其渗出点个数,并采用多元统计分析对数据进行处理。结果:①镜下空白组大鼠结肠黏膜结构清晰;各模型组可见结肠黏膜水肿及炎性细胞浸润。②腹部EB渗出点的聚类分析结果表明其特征网格内的渗出点无时间变化趋势(P>0.05);且各时间点渗出点分布与浓度变化无明显相关性(P>0.05)。③7.5%浓度各时间组腹部EB渗出点因子分析及轮廓分析结果表明,特征区域Ⅱ渗出趋势随时间变化显著(P<0.05),且渗出点位置相当于"天枢"穴附近。④造模后5 h各浓度组腹部EB渗出点因子分析及轮廓分析结果表明,"区域C"渗出程度随浓度变化显著(P<0.05),且渗出点位置亦相当于"天枢"穴附近。结论:基于多元统计分析方法能进一步明确AIMI大鼠EB渗出点的特征区域,并发现该特征区域存在与时间及病情程度相关的穴位分布特征。

Objective To observe the distribution characters of Evans Blue(EB) exudation spots in the abdominal area in acute intestinal mucosal injury(AIMI) rats by using latitude and longitude grid counting and multivariate statistical analysis and to explore the feasibility of these methods. Methods Twenty-four SD rats were randomly divided into blank control, 7.5%, 10%, and 12.5% mustard oil groups(n=6 rats in each group). The AIMI model was established by mustard oil enema, followed by injection of EB(0.1 mL/100 g) into the tail vein. At 5, 10, 15, 20 and 25 h after EB injection, the rats under anesthesia were fixed in supine position for observing and photographing the abdominal subcutaneous EB exudation spots. The H.E. staining was used to observe histopathological changes of colonic mucosa. The longitude and latitude grids of the abdominal region were constructed(by taking the midpoint of the superior sternum as the origin) to determine the position of the blue spots. That the coordinate grids of the two regions can be connected geometrically is termed as "characteristic region". The data were processed by using multivariate statistical analysis. Results ① H.E. staining showed edema and inflammatory cell infiltration after colonic enema of different concentrations of mustard oil. ② Clustering analysis indicated that the distribution of exudation points in the "characteristic grid" had no temporal variation trend, and was not related with the concentration of mustard oil(P>0.05). ③ Factor analysis and contour analysis about the exudation spots of EB at 7.5% concentration showed that the "characteristic region Ⅱ" of different factors presented a tendency of time-dependent exudation, i.e. reduction of exudation degree along with time(P<0.05), and it is located near "Tianshu" acupoint. ④At 5 h after injection of EB, the 8 "characteristic regions" presented an EB-concentration-dependent tendency(reduction in exudation degree along with the increase of EB-concentration), among which the exudation degree of region C(near "Tianshu" acupoint) of the abdomen was higher(P<0.05). Conclusion The multivariate statistical analysis method can be used to identify the abdominal "characteristic regions" of exudation spots of EB in rats with AIMI, and the characteristic region has acupoint sensitization characteristics related to the time and severity of mucosal injury.