形态学指标测量与人脾破裂损伤时间推断的观察

Observation of morphological indexes measurement and wound age estimation in human spleen rupture

目的观察人脾破裂和大鼠脾锐器创损伤后不同时间形态学指标的变化，寻求推断损伤时间的形态学方法和分子指标。方法选取118例外伤性脾破裂标本，按破裂损伤时间分为10组；建立大鼠脾锐器创模型。HE染色后分别观察人和大鼠脾破裂损伤中央区、纤维素渗出带和损伤外周区的组织形态学变化；采用免疫组化和图像分析法检测人脾破裂损伤中央区、纤维素渗出带和损伤外周区MPO、CD68、α-SMA阳性细胞的数量变化。结果脾破裂损伤组织形态学变化经历炎症期、增生期、肉芽组织及瘢痕组织形成期。免疫组化标记示MPO阳性细胞数在脾伤后6-12h开始升高，12-24h阳性细胞数最多，随后逐渐下降，0～5天相邻各损伤组之间比较差异均有统计学意义（P〈0．05）；CD68阳性细胞数在伤后12-24h开始升高，第7天阳性细胞数最高，之后开始下降，12～24h以后相邻各损伤组之间比较差异均有统计学意义（P〈0．05）；α-SMA在损伤24h之内无阳性表达，第2天开始出现少量表达，随后依次升高，损伤24h后各损伤组之间差异均有统计学意义（P〈0．05）。结论MPO、CD68及α-SMA在脾破裂损伤后组织中表达呈时序性改变；多指标综合应用结合脾破裂组织形态学改变有望更加精确推断人脾破裂损伤时间。

Purpose To observe the changes of morphological indexes in human spleen rupture and sharp injury of rat spleen at differ- ent intervals, and then to find out morphological methods and molecular indexes for estimating wound age. Methods 118 cases of traumatic spleen rupture specimens were selected and divided into 10 groups according to the rupture time; the sharp injury spleen model of rat was established. The morphological changes of central area, fibrin exudation zone and peripheral area of spleen rupture can be observed using HE staining; changes of the number of MPO, CD68 and et-SMA positive cells around the central area, fibrin exuda- tion zone and peripheral area of human spleen rupture can be observed using immunohistochemistry staining and image analysis method. Results Morphological characteristics of spleen rupture have experienced three changing stages： inflammatory stage, the proliferative stage, granulation tissue and scar tissue formative stage. The result of immunostaining revealed that MPO-positive cells in 6 - 12 hours began to increase, reached their peak within 12 - 24 hours and then declined gradually. And the differences of the adjacent injury groups during the injury time 0 - 5 days were statistically significant （P 〈 0.05 ）. CD68-positive cells began to increase in 12 - 24 hours group, reached the highest number of positive cells on the 7 days and then declined. And the differences of the adjacent injury groups after the 12 - 24 hours group were statistically significant （P 〈 0.05 ）. α-SMA found no expression within 24 hours of injury, maintained at a low level of α-SMA on the 2 days and then increased gradually. After 24 hours of injury, the differences of the adjacent injury groups were statistically significant （P 〈 0.05）. Conclusions Spleen tissue after injury can induce a chronological expression of MPO, CD68 and α-SMA. The results can be hoped that it is more accurate for age estimation of human spleen rupture using muhi- indexes in combination with morphological characteristics of spleen rupture.