Effect of Bushenantai Recipe on the Expression of Endometrial LIF in Mice with Embryonic Implantation Dysfunction

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were randomized into three groups： normal control group, model group and traditional Chinese medicine group （TCM group） （n=40 in each group）. Uterus was collected on the pregnancy day （Pd） 4, 5, 6 after an intravenous injection of Evan＇s blue. The endometrium was dyed by Evan＇s blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.

Mechanisms of Traditional Chinese Medicine Bushenantai granules(补肾安胎颗粒) in promoting angiogenesis at the maternal-fetal interface of recurrent spontaneous abortion mice

OBJECTIVE:To assess the effects of Bushenantai(BSAT)granule(补肾安胎颗粒)on angiogenesis-related factors[E2,P,and vascular endothelial growth factor(VEGF)]at the maternal-fetal interface of recurrent spontaneous abortion(RSA)mice,and to evaluate the role of BSAT in promoting angiogenesis at the maternal-fetal interface by influencing the expression of sex hormones,and VEGF.METHODS:A mouse model with normal pregnancy and another with Clark’s classic RSA were established.The RSA mice were randomly assigned to six groups:normal,model,progesterone,high-doseBSAT granule(BSAT-H),medium-dose-BSAT granule(BSAT-M),and low-dose-BSAT granule(BSAT-L)(n=10 for each group).The embryo loss rate and the histopathological changes in the decidual tissues were measured.Serum levels of estrogen(E2),progesterone(P),and VEGF were detected by enzyme-linked immunosorbent assay.The m RNA and protein expressions of estradiol receptor(ER),progesterone receptor(PR),VEGF,and vascular endothelial growth factor receptor 2(VEGFR2)in the decidual tissues were identified by immunohistochemistry,Western blotting,and quantitative reverse transcription polymerase chain reaction.RESULTS:The embryo loss rate in all groups that received BSAT treatment was reduced,while the number of blood vessels at decidual tissues was increased.The serum levels of E2,P and VEGF were elevated,and the m RNA and protein expressions of ER,PR,VEGF,and VEGFR2 in the decidual tissues were enhanced.CONCLUSION:BSAT can improve angiogenesis at the maternal-fetal interface and reduce the embryo loss rate,which may be associated with its ability to increase the serum levels of estrogen,progesterone,and VEGF,in addition to up-regulation of m RNA and protein expression of ER,PR,VEGF,and VEGFR2 in the decidual tissues.

补肾安胎方对胚泡着床障碍小鼠子宫内膜HB-EGF及其受体EGFR表达的影响

目的观察中药补肾安胎方对胚泡着床障碍模型小鼠胚泡着床情况及对类肝素结合样表皮生长因子(HB-EGF)及其受体(EGFR)表达的影响。方法将妊娠雌鼠随机分为正常组、模型组、中药组,每组30只,分别于妊娠第5、6天(Pd5、6)用伊文思蓝尾静脉注射后取妊娠子宫。观察妊娠第5天(Pd5)各组对伊文思蓝的着色反应;运用RT-PCR方法检测HB-EGFmRNA的含量,采用免疫组织化学法测定HB-EGF及其受体EGFR的表达。结果Pd5,模型组对伊文思蓝反应和平均反应点数显著低于正常组,中药组与模型组有显著差异。Pd5、6,模型组子宫内膜HB-EGF及其受体EGFR的表达在时间上明显滞后于正常组并且表达量减少,中药组与模型组有显著差异(P<0.05)。结论补肾安胎方改善胚泡着床障碍小鼠胚胎着床可能与提高着床位点的HB-EGF及其受体EGFR的表达有关。

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

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。

补肾安胎方及其拆方对控制性超排卵小鼠胚胎着床干预的比较

目的观察补肾安胎方及其补肾、活血组分对控制性超排卵(controlled ovarian hyperstimulation,COH)小鼠胚泡围着床期类肝素结合表皮生长因子(heparin-binding epidermal growth factorlike growthfactor,HB-EGF)及其受体的影响,探讨补肾安胎方及其拆方对着床环节的作用机制。方法将雌性小鼠随机分为正常组(N)、模型组(M)、补肾安胎组(BH)、补肾组(BS)和活血组(HX)。观察妊娠第1天阴栓率、妊娠第6天妊娠率和孕鼠着床位点数;采用反转录聚合酶链反应(RT-PCR)方法测定妊娠第5天、第6天子宫内膜HB-EGF mRNA的表达,采用免疫组织化学法测定同期子宫内膜表皮生长因子受体(epidermal growth factor receptor,EGFR)蛋白的表达。结果模型组阴栓率、妊娠率较正常组显著降低(P<0.01),着床位点数显著升高(P<0.01);中药各组治疗后,阴栓率、妊娠率均较模型组显著升高(P<0.05,P<0.01),但着床位点数均显著降低(P<0.05,P<0.01),以补肾安胎组作用最为明显;活血组阴栓率高于补肾组(P<0.05),着床位点数较补肾组降低(P<0.01),妊娠率两组比较,差异无统计学意义。正常组HB-EGF mRNA表达最高,模型组显著低于正常组(P<0.01),各中药组均显著高于模型组(P<0.01);各中药组HB-EGF mRNA表达比较,补肾安胎组明显高于活血组和补肾组(P<0.05),活血组高于补肾组(P<0.05);妊娠第6天,HB-EGF mRNA表达量较妊娠第5天降低。EGFR蛋白在围着床期子宫内膜腺体和基质细胞胞质中表达,各组小鼠EGFR蛋白表达空间分布差异无统计学意义。正常组EGFR蛋白表达丰富;模型组表达较正常组显著减少(P<0.01),部分缺失;中药各组EGFR表达量均高于模型组(P<0.01);其中补肾活血组高于活血组和补肾组(P<0.05,P<0.01),活血组高于补肾组(P<0.05,P<0.01)。结论补肾安胎方及补肾、活血组分均能够改善控制性超排卵小鼠的着床率和妊娠率,促进胚泡生长和HB-EGF及其受体的表达,其中以补肾安胎全方作用最佳,补肾、活血组分在改善胚泡着床过程中具有协同作用。

补肾安胎法治疗复发性自然流产的思考与运用

复发性自然流产严重影响了妇女的生殖健康和生活质量。本病病因复杂,机制尚未明确,中医理论认为本病责之于肾虚,以补肾安胎法为基本方法治疗本病,多取得满意疗效,值得临床推广。笔者总结近5年补肾安胎法的实验及临床研究,思考补肾安胎治疗复发性流产的存在问题及研究前景。

补肾安胎方联合黄体酮治疗复发性流产的疗效分析

目的 探讨黄体酮联合补肾安胎方治疗抗心磷脂抗体阳性复发性流产的临床效果.方法 选取抗心磷脂抗体阳性复发性流产患者98例,采用随机数字表法分为观察组和对照组各49例,对照组采用黄体酮胶丸治疗,观察组在对照组的基础上联合补肾安胎方治疗.比较两组的治疗时间、活产率和抗体清除率.结果 观察组治疗时间（179.5±12.8）d,明显短于对照组的（288.6±14.5）d,差异有统计学意义（t=6.249,P ＜0.05）;观察组活产率（91.8％）、抗体清除率（79.6％）明显高于对照组（79.6％、36.7％）,差异均有统计学意义（χ2=5.247、4.998,均P＜0.05）;观察组发生不良反应3例（6.0％）,对照组发生不良反应15例（30.6％）,两组不良反应发生率差异有统计学意义（χ2=8.250,P＜0.05）.结论 黄体酮联合补肾安胎方治疗抗心磷脂抗体阳性复发性流产可缩短治疗时间,改善妊娠结局,提高抗体清除率.