解郁育胞方对慢性应激致大鼠子宫内膜低容受性的机制研究

Effect of Jieyuyubao Prescription on endometrial receptivity of the chronic stress rats

目的:研究解郁育胞方对慢性应激大鼠妊娠率、平均着床点数的影响,以及对血清β-内非肽(β-EP)、子宫内膜血管内皮生长因子(VEGF)及胞饮突的影响,探讨其对慢性应激大鼠子宫内膜容受性的调节机制。方法:80只SPF级SD大鼠按体重分层法随机分为5组:正常对照组、应激组、应激+5.6g/kg解郁育胞方组、应激+11.2g/kg解郁育胞方组、应激+22.4g/kg解郁育胞方组。除正常对照组外,其余4组大鼠建立慢性应激模型,于刺激第15日开始,应激+11.2g/kg解郁育胞方组每日9:00给予解郁育胞方溶液(相当于生药1g/ml)灌胃,给药容量为11.2g/kg;正常对照组、应激组灌服等体积纯化水(11.2g/kg);应激+5.6g/kg解郁育胞方组、应激+22.4g/kg解郁育胞方组分别按应激+11.2g/kg解郁育胞方组折算临床有效剂量的1/2和2倍给药,给药容量5.6g/kg和22.4g/kg,每日1次。5组大鼠分别于第28日16:00按照雌:雄=2:1合笼,经阴道涂片确认妊娠后第4日停药,当日16:00处死大鼠,检测血清β-EP、子宫内膜VEGF含量及子宫内膜胞饮突成熟度。结果:平均妊娠率和着床点数:各组经过10日合笼,应激组和应激+5.6g/kg解郁育胞方组各有1只大鼠仍未妊娠;与应激组相比,应激+11.2g/kg解郁育胞方组和应激+22.4g/kg解郁育胞方组的平均着床点数明显增高。2血清β-EP含量:与正常对照组相比,应激组、应激+5.6g/kg解郁育胞方组的β-EP含量明显升高;应激+11.2g/kg解郁育胞方组、应激+22.4g/kg解郁育胞方组与应激组比较有显著差异;应激+11.2g/kg解郁育胞方组组、应激+22.4g/kg解郁育胞方组的β-EP含量明显低于应激+5.6g/kg解郁育胞方组。VEGF的表达:VEGF主要位于子宫内膜腺上皮细胞的胞浆和部分基质血管内皮细胞的胞浆,部分间质细胞有弱着色,各试验组表达强弱不一。与A正常对照组相比,应激组VEGF的表达明显降低;应激+22.4g/kg解郁育胞方组与应激组相比有显著统计学意义。4透射电镜子宫内膜上皮细胞胞饮突表达:与正常对照组相比,应激组、应激+5.6g/kg解郁育胞方组评分明显降低;与应激组比较,应激+11.2g/kg解郁育胞方组、应激+22.4g/kg解郁育胞方组的胞饮突评分明显高于应激组;应激+11.2g/kg解郁育胞方组、应激+22.4g/kg解郁育胞方组的胞饮突评分明显高于应激+5.6g/kg解郁育胞方组。结论:解郁育胞方能有效的改善或降低慢性应激大鼠血清β-EP值,降低应激对机体的损伤,增加机体抗应激能力,且呈现一定剂量依赖性。2解郁育胞方高剂量能明显提高慢性应激大鼠子宫内膜中VEGF表达水平以及子宫内膜上皮细胞的胞饮突成熟度。我们推测,解郁育胞方通过平衡体内由于紧张应激导致的β-EP分布,提高子宫内膜VEGF表达水平及内膜上皮细胞的胞饮突成熟度,从而改善子宫内膜容受性,增加大鼠妊娠着床点数。

Objective： To study the impact of Jieyuyubao prescription in rats with chronic stress for serum β-EP,endometrial VEGF and pinopodes expression,revealing the chronic stress rats on endometrial receptivity regulatory mechanisms. Methods： 80 SPF SD rats were stratified randomly into 5 groups： normal control group,stress group and stress ＋ 5. 6g / kg Jieyuyubao prescription solution group,stress ＋ 11. 2g / kg Jieyuyubao prescription solution group,stress ＋ 22. 4g / kg Jieyuyubao prescription solution group. In addition to the normal control group,the remaining four groups of rats were established drug chronic stress model. stress ＋ 11. 2g / kg Jieyuyubao prescription solution group daily 9： 00 give Jieyuyubao prescription solution（ equivalent to crude drug 1g / ml） by gavage,dosing capacity for 11. 2g / kg; a normal control group,stress group filling service such as volume purified water（ 11. 2g / kg）; stress ＋ 5. 6g / kg Jieyuyubao prescription solution group and stress ＋22. 4g / kg Jieyuyubao prescription solution group were in stress ＋ 11. 2g / kg Jieyuyubao prescription solution group conversion clinically effective doses of 1 /2 and 2 times to medicine and drug delivery capacity 5. 6g / kg and 22. 4g / kg,daily 1 times. Five groups of rats respectively in the 28 th 16： 00 in accordance with the female： male = 2： 1） caged and the vaginal smear pregnancy is confirmed after the 4th withdrawal.At 16： 00 the same day killed rats,serum β-EP,uterine endometrial VEGF levels and endometrial pinocytic process maturity. Results： The average pregnancy rate and the number of implantation： 10 rats in each group were not pregnant at each group of 1 rats. The average number of implantation points in group stress ＋ 22. 4g / kg Jieyuyubao prescription solution group and stress ＋ 11. 2g / kg Jieyuyubao prescription solution group was significantly higher than that in stress ＋ 5. 6g / kg Jieyuyubao prescription solution group（ P 0. 01）. the serum levels of beta endorphine： A and compared to the normal control group,stress group and stress ＋ 5. 6g / kg Jieyuyubao prescription solution group,the levels of beta endorphine increased significantly（ P 0. 05）; stress ＋ 11. 2g / kg Jieyuyubao prescription solution group,stress ＋ 22. 4g / kg Jieyuyubao prescription solution group and stress group compared with significant difference（ P 0. 05）; stress ＋ 11. 2g / kg Jieyuyubao prescription solution group,the β-EP content was significantly lower than that ofstress ＋ 5. 6g / kg Jieyuyubao prescription solution group（ P 0. 05）. The expression of VEGF： VEGF was mainly located in the cytoplasm of endometrial glandular epithelial cell cytoplasm and some stromal vascular endothelial cells and some stromal cells had weak coloring. The test group expression strength. Compared with the normal control group,the expression of VEGF in B group was significantly lower than that in normal control group（ P 0. 05）,and stress ＋ 22. 4g /kg Jieyuyubao prescription solution groupwas significantly significantly（ P 0. 05） compared with the stress group. 4Transmission electron microscopy of endometrial epithelial cells pinopod expression： compared with the normal group of a,stress group,the score of stress ＋ 11.2g / kg Jieyuyubao prescription solution group decreased significantly（ P 0. 05）; compared with the stress of stress ＋ 5. 6g / kg Jieyuyubao prescription solution group,stress ＋ 11. 2g / kg Jieyuyubao prescription solution groupand stress ＋ 22. 4g / kg Jieyuyubao prescription solution groupof pinopodes score was significantly higher than that of stress group（ P 0. 05）; stress ＋ 11. 2g / kg Jieyuyubao prescription solution group,stress ＋ 22. 4g / kg Jieyuyubao prescription solution group pinopodes score was significantly higher than that of stress ＋ 5. 6g / kg Jieyuyubao prescription solution group（ P 0. 05）. Conclusion： the effective improvement of the serum levels of β-EP in rats with depression can decrease the damage to the body and increase the anti stress ability of the body,and it will be dose dependent. The expression levels of VEGF in the endometrium of rats with chronic stress and the maturity of the cytoplasm of the endometrial epithelial cells could be significantly increased by the high dose of the rats.. We hypothesized that Jieyuyubao prescription by balancing the body due to the stress caused byβ-EP distribution,improve the endometrial VEGF expression level and endometrial epithelial cells of pinopodes maturity,so as to improve the endometrial receptivity.