软坚散结中药对子宫内膜异位症模型小鼠内膜黏附因子的影响

Effect of Herbs Softening and Resolving Masses on the Adhesion Factors of Endometrium of Endometriesis Model Mice

目的观察软坚散结中药对子宫内膜异位症模型小鼠异位内膜黏附的影响。方法用增强型绿色荧光蛋白(EGFP)转基因和野生型C57BL/6J小鼠建立子宫内膜异位症模型,中药组给予软坚散结中药灌胃,空白组、模型对照组(对照组)则给予等量的生理盐水灌胃,2周后成像并比较两组小鼠荧光表达的强弱;流式细胞术检测异位病灶的凋亡情况;W estern b lot和实时定量PCR等技术对异位内膜的细胞间黏附分子1(ICAM-1)和透明质酸受体CD 44进行检测。结果镜下可见模型小鼠肠管间、腹壁下、肝小叶下或脾周形成绿色团块状或星点状内膜异位病灶,中药组异位灶体积及荧光表达强度明显低于对照组(P<0.05),中药组和对照组异位内膜细胞的凋亡率与空白组差异有统计学意义(均P<0.01),且中药组高于对照组(P<0.05);中药组CD 44、ICAM-1蛋白和mRNA表达显著低于模型对照组(均P<0.01)。对照组、中药组的CD 44和ICAM-1mRNA表达量分别是空白组的16.34、2.75和1.53、1.19倍。结论软坚散结中药可抑制子宫内膜异位症模型小鼠异位内膜病灶的产生,这种作用可能是通过降低黏附因子ICAM-1、CD 44的表达而实现的。

Objective To evaluate the effect of herbal medicines with the functions to soften and resolve masses of ectopic endometrium of mouse model of endometriosis. Methods The enhanced green fluorescent protein （EGFP） transgenic and wild-type C57 BL/6J mice were used to establish the mouse model of endometriosis. For the herb group, the herbal medicine was administered by gavage. For the control and blank groups, the same amount saline was given by gavage. Two weeks later, all model mice were sacrificed and optical density of fluorescence of ectopic endometrium was calculated for comparison. The ectopic endometrium, uterus and ovaries were examined histologically and FCM analysis was performed to access cell apoptosis, Western blot and Real-time PCR was performed to access the expression of ICAM-1 and CD44 in the ectopic endometrium. Results Under fluorescent microscope, the green cake-like or punctiform ectopic endometrium were found between the intestines, under abdominal wall or under hepatic lobules or around spleen in the model mice. The volume and optical density of fluorescence of ectopic endometrium of the herb group were significantly lower than those of the control group （P 〈 0. 05）, the cell apoptosis of the herb group and the control group was significantly different from that of the blank group （P 〈 0.01）, and the herb group was also higher than the control group （P〈0. 05）. The expression of CD44, ICAM-1 and mRNA in the herb group was significantly lower than that of the control group （P 〈 0. 01）. The expression of CD44, ICAM-1 and mRNA of the control and the herb group was 16.34 and 2. 75, and 1.53 and 1.19 times respectively of the blank group. Conclusion The herbs with the functions to soften and resolve masses exert a growth-inhibitory effect on ectopic endometrium of model mice. It is may be realized by inhibiting the expression of CD44 and ICAM-1.