补肾安胎方改善胚胎着床障碍小鼠着床局部血管形成及其机制

Bushenantai Recipe Improving Angiogenesis at Implantation Site in Mice with Embryonic Implantation Dysfunction and the Mechanism

目的观察补肾安胎方对吲哚美辛诱导着床障碍小鼠着床局部血管形成的影响并探讨其机制。方法将妊娠小鼠随机分为正常组、吲哚美辛组(模型组)、补肾安胎方组(中药组)。正常组于妊娠第3、4天每天2次皮下注射溶剂,模型组和中药组均于妊娠第3、4天每天2次皮下注射吲哚美辛(4.33 mg/kg)造成小鼠胚胎着床障碍。自妊娠第1天开始,正常组和模型组于每日上午灌服自来水0.3 ml,中药组则灌服中药0.3 ml。比较3组小鼠在妊娠第8天妊娠率、妊娠小鼠胚胎着床位点数;同时,取小鼠子宫,进行苏木精-伊红染色及用免疫组化技术观察CD34、血管内皮生长因子(VEGF)及其受体表达。结果模型组小鼠妊娠率和胚胎着床位点数(27.3%、5.3±0.7)显著低于正常组(90.9%、13.3±2.8)(均P<0.01);中药组小鼠妊娠率和胚胎着床位点数(72.7%、10.7±2.2)较模型组显著增高(均P<0.05)。苏木精-伊红染色及CD34免疫组织化学染色结果显示:正常组血管丰富,模型组血管稀少,中药组血管较模型组丰富,与正常组相当。免疫组织化学结果显示,妊娠第8天正常组子宫内膜中VEGF及其受体表达明显,模型组中的表达显著减少,中药组介于两组之间。结论补肾安胎方提高吲哚美辛诱导着床障碍小鼠妊娠率和胚胎着床位点数,其机制可能是通过改善子宫内膜VEGF及其受体的表达有关。

Objective To investigate the effect of Bushenantai recipe on pregnancy rate and number of implanted embryo of mice with embryonic implantation dysfunction induced by indomethacin and its mechanism.Methods All pregnancy mice were randomly divided into 3 groups: normal group,indomethacin group(model group) and Bushenantai recipe group(CHM group).The mice in the normal group was injected subcutaneously with gingili twice on the pregnancy day 3 and 4,while indomethacin(4.33 mg/kg) was administrated subcutaneously twice on the pregnancy day 3 and 4 to establish animal models of embryonic implantation dysfunction in model group and CHM group.The pregnancy rate and number of implanted embryo were compared among 3 groups on the 8th day of pregnancy,while the uteri were dyed by hematoxylin and eosin(HE),and the expression of the CD34,vascular endothelial growth factor(VEGF) and its receptor-1(VEGFR-1) in endometrium was stained by immunohistochemistry.Results The pregnancy rate and number of implanted embryo were significantly lower in indomethacin group(27.3% and 5.3±0.7) than in normal group(90.9% and 13.3±2.8)(both P<0.01),but was higher in CHM group(72.7% and 10.7±2.2) than in indomethacin group(both P<0.05).HE staining and immunohistochemistry for CD34 revealed that endometrial blood vessels were abundant in normal group,but less in indomethacin group,and those in CHM group were more than those in indomethacin group,and similar with normal group.Immunohistochemistry revealed that VEGF and VEGFR on the 8th day in endometrium of mice were significantly decreased in indomethacin group and CHM group as compared with normal group.Conclusion Bushenantai recipe could increase pregnancy rate and number of implanted embryo of mice with embryonic implantation dysfunction induced by indomethacin,which might be contributed to the regulation of the expression of VEGF and VEGFR in endometrium.