妇痛宁对子宫内膜异位症COX2-PGE2作用途径的影响

Effect on the COX2-PGE2 pathways of endometriosis by Fu Tong Ning

[目的]通过研究化瘀散结中药妇痛宁对子宫内膜异位症(EMs)病灶中环氧化酶2(COX2)-前列腺素E2(PGE2)作用途径中相关分子基因和蛋白的表达,探讨妇痛宁对EMs中血管生成、侵袭和转移方面的影响。[方法]收集EMs患者的异位病灶组织,原代消化培养子宫内膜细胞,将25×106/m L浓度混合培养的异位子宫内膜细胞注射到裸鼠腹部皮下,成功建立模型后,将成模裸鼠分为空白对照组、低剂量组、中剂量组、高剂量组和阳性对照组,分别予无菌生理盐水、低剂量中药、中剂量中药、高剂量中药和孕三烯酮灌胃,3周后处死裸鼠,取病灶,测量病灶体积,免疫组化法检测各组病灶中血管内皮生长因子(VEGF)的表达和微血管密度(MVD),逆转录-聚合酶链反应(RT-PCR)法检测病灶中COX2、PGE2、VEGF、Snail和E-cadherin m RNA的表达,Western-blot法检测病灶中Snail和E-cadherin蛋白的表达。[结果]中剂量及高剂量妇痛宁治疗组裸鼠异位病灶体积减小、VEGF表达及MVD均降低,与空白对照组比较差异有统计学意义(P<0.05)。各剂量治疗组裸鼠异位病灶中COX2、PGE2、VEGF、Snail m RNA表达降低,Ecadherin m RNA表达升高,且Snail蛋白表达降低,E-cadherin蛋白表达升高,与空白对照组比较差异有统计学意义(P<0.05)。[结论]妇痛宁可影响COX2-PGE2作用途径中与血管生成、侵袭和转移相关分子的表达,通过抑制异位病灶的血管生成、侵袭和转移能力而抑制病灶的生长。

[Objective] To study the expressions of related molecules gene and protein in COX2-PGE2 pathways of endometriosis lesions by removing blood stasis and dispersing phlegm Chinese medicine, investigate the effects on angiogenesis, invasion and metastasis of EMs by Fu Tong Ning. [Methods] Ectopic endometrial samples were obtained and endometrial cells were cultured respectively. The concentration of 25×106m L co-cultured ectopic endometrial cells of EMs patients were injected into the abdominal subcutaneous tissue of nude mice. After the models were established succeffuly, the model mice were randomly divided into the blank control group, lower dose treatment group, intermediate dose treatment group, high dose treatment group and positive control group. The mice were given sterilized saline solution, lower dose Fu Tong Ning Chinese medicine, intermediate dose Fu Tong Ning Chinese medicine, high dose Fu Tong Ning Chinese medicine and gestrinone capsules respectively. Mice were euthanized after three weeks and taken the lesions. The volumes of lesions were calculated. The expressions of VEGF and CD34 were detected by immunohistochemical examination, then the MVD was calculated. The expressions of COX2, PGE2, VEGF, Snail and E-cadherin m RNA in each group were detected by RT-PCR. The expressions of Snail and E-cadherin protein were detected by Western-blot. [Results] The volumes of lesions were smaller, the expressions of VEGF and MVD were decreased in intermediate and high dose treatment groups, there were statistically significant differences compared with the blank control group(P<0.05). The expressions of COX2, PGE2, VEGF and Snail m RNA were decreased,but the expressions of E-cadherin m RNA were increased in each treatment groups, meanwhile. The expressions of Snail protein were decreased and the expressions of E-cadherin protein were increased in each treatment groups, and there were statistically significant differences compared with the blank control group(P<0.05). [Conclusion] Fu Tong Ning Chinese medicine could affect the expression of molecules related to angiogenesis, invasion and metastasis in COX2-PGE2 pathways, and the growth of the lesions were restrained by inhibiting the ability of angiogenesis, invasion and metastasis.