穴位组织切片中肥大细胞分布和激活的定量观察

Quantitative Observation of the Distribution and Activation of Mast Cells in Tissue Samples of Acupoints

肥大细胞是穴位功能的重要载体,为实现对组织中肥大细胞启动情况的定量观察,提出了一种计算机辅助方法对组织的平均肥大细胞脱颗粒率进行计算。该方法根据甲苯胺蓝染色中出现的特异性深紫色染色对肥大细胞进行定位,根据形态学特征选择了胞内空泡面积、胞外颗粒数和周长面积比三个参数作为激活特征参数,提出了脱颗粒率计算方法。通过对急性佐剂型关节炎大鼠针刺后足三里穴位组织进行的观察,发现该方法的肥大细胞识别率较高,并检出针刺导致肥大细胞脱颗粒率明显升高(与模型组比较,P<0.01,n=697~858)。结果证明,该计算机辅助方法对肥大细胞激活情况的定量观察可行、有效。

Mast cells play important roles in the function of acupoints. In order to quantitatively observe the activation of mast cells in tissues, we proposed a computer - aided method to calculate the average mast cell de-granulation ratio in tissues. Skin tissues with acupoints from rats were processed into paraffin sections and dyed with toluidine blue, which could well locate mast cells. According to the morphological descriptions of mast cells degranulations, three characteristic parameters, including the area of intracellular cavity, number of extracellular granule and ratio of cell circumference/area, were measured for the characterization of mast cells degranulations. The average values were used as thresholds, cells with at least 2 exceeding parameters were determined as degranu- lated. The tissue of ST 36 from acute adjuvant induced arthritis rats were examined with this method. In a test for recognition, 499 of 501 mast cells were successfully detected. Subjective examinations had confirmed that acu-puncture could induce mast cell degranulation in the acupoint. Additionally, aided with this new method, we have achieved the same results in objective analysis （P 〈 0. 01, vs Model group, n = 697 - 858 ） . These results show that this computer - aided method is practical and valid for quantitative analysis of mast cellos activation.