补肾化瘀方对多囊卵巢模型大鼠子宫内膜胞饮突及ER、PR蛋白表达的影响
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摘要
目的:观察补肾化瘀方对多囊卵巢模型大鼠子宫内膜胞饮突及雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)蛋白表达的影响。方法:100只SD大鼠随机选取30只为空白对照组;另70只为模型制备组,均以颈背部皮下注射脱氢表雄酮(dehydroepiandrostexone,DHEA)溶液建立多囊卵巢模型大鼠。造模成功后,随机分为模型对照组、达英-35组、补肾化瘀方组,分别给予蒸馏水、达英-35及补肾化瘀方对大鼠灌胃给药,连续灌胃21 d,停药7 d为1个周期,3个周期后停药。按雌雄鼠11合笼,并于妊娠第4天处死大鼠。采用扫描电镜观察各组多囊卵巢模型大鼠子宫内膜胞饮突表达,免疫组化法检测大鼠子宫内膜ER、PR蛋白的表达。结果:补肾化瘀方可提高多囊卵巢模型大鼠胞饮突的表达,与空白对照组、模型对照组、达英-35组比较,补肾化瘀方组大鼠子宫内膜表面微绒毛低矮,局部消失,可见大量胞饮突,表面光滑,形态基本规则,边界清楚。ER表达:与空白对照组比较,模型对照组、达英-35组、补肾化瘀方组分别0.491±0.020、0.379±0.014、0.383±0.019,高于空白对照组,差异有显著统计学意义(P<0.01);与模型对照组比较,达英-35组、补肾化瘀方组低于模型对照组,差异具有显著统计学意义(P<0.01)。PR表达:与空白对照组比较,模型对照组、补肾化瘀方组分别为0.292±0.02、0.215±0.012,显著高于空白对照组,差异有统计学意义(P<0.01),达英-35组为0.210±0.014,高于空白对照组,差异有统计学意义(P<0.05);与模型对照组比较,达英-35组、补肾化瘀方组低于模型对照组,差异有统计学意义(P<0.05)。结论:补肾化瘀方可提高多囊卵巢模型大鼠子宫内膜胞饮突的表达,降低ER、PR蛋白水平,这可能是其改善子宫内膜容受性的机制之一。
Objective:To observe the effects of Bushen Huayu Recipe on endometrial cytoplasm and estrogen receptor(ER) and progesterone receptor(PR) protein expression in polycystic ovary model rats.Methods:Thirty SD rats were randomly selected from the blank control group.The other 70 rats were model preparation groups.Polycystic ovary model rats were established by subcutaneous injection of dehydroepiandrostexone(DHEA) solution.After successful modeling,they were randomly divided into model control group,Daying-35 group,and Bushen Huayu Recipe group.Distilled water,Daying-35 and Bushen Huayu Recipe were administered to rats for intragastric administration for 21 days.The drug was stopped for 7 days,and the drug was stopped after 3 cycles.The male and female rats were 1:1 caged,and the rats were sacrificed on the fourth day of pregnancy.The expression of ER and PR protein in endometrium of rats in polycystic ovary model rats were observed by scanning electron microscopy.The expression of ER and PR protein in rat endometrium was detected by immunohistochemistry.Results:Bushen Huayu Recipe could increase the expression of pinocytosis in rats with polycystic ovary model.Compared with the blank control group,model control group and Daying-35 group,the rat endometrial surface microvilli in Bushen Huayu Recipe group was low,partially disappeared,with a large number of pinocytosis,smooth surface,basic shape and clear boundaries.ER expression:Compared with the blank control group,the model control group,Daying-35 group,and Bushen Huayu Recipe group were 0.491±0.020,0.379±0.014,and 0.383±0.019,respectively,which was higher than the blank control group,and the difference was statistically significant(P<0.01);Compared with the model control group,the Da Ying-35 group and the Bushen Huayu Recipe group were lower than the model control group,and the difference was statistically significant(P<0.01).PR expression:Compared with the blank control group,the model control group and the Bushen Huayu Recipe group were 0.292±0.02 and 0.215±0.012,respectively,which were significantly higher than the blank control group,and the difference was statistically significant(P<0.01).The 35 groups were 0.210±0.014,which was higher than the blank control group,and the difference was statistically significant(P<0.05).Compared with that the model control group,the Daying-35 group and the Bushen Huayu recipe group were significantly lower than the model control group(P<0.05).Conclusion:Bushen Huayu Recipe can improve the expression of endometrial pinocytosis and decrease the levels of ER and PR protein in polycystic ovary model rats,which may be one of the mechanisms to improve endometrial receptivity.
Objective:To observe the effects of Bushen Huayu Recipe on endometrial cytoplasm and estrogen receptor(ER) and progesterone receptor(PR) protein expression in polycystic ovary model rats.Methods:Thirty SD rats were randomly selected from the blank control group.The other 70 rats were model preparation groups.Polycystic ovary model rats were established by subcutaneous injection of dehydroepiandrostexone(DHEA) solution.After successful modeling,they were randomly divided into model control group,Daying-35 group,and Bushen Huayu Recipe group.Distilled water,Daying-35 and Bushen Huayu Recipe were administered to rats for intragastric administration for 21 days.The drug was stopped for 7 days,and the drug was stopped after 3 cycles.The male and female rats were 1:1 caged,and the rats were sacrificed on the fourth day of pregnancy.The expression of ER and PR protein in endometrium of rats in polycystic ovary model rats were observed by scanning electron microscopy.The expression of ER and PR protein in rat endometrium was detected by immunohistochemistry.Results:Bushen Huayu Recipe could increase the expression of pinocytosis in rats with polycystic ovary model.Compared with the blank control group,model control group and Daying-35 group,the rat endometrial surface microvilli in Bushen Huayu Recipe group was low,partially disappeared,with a large number of pinocytosis,smooth surface,basic shape and clear boundaries.ER expression:Compared with the blank control group,the model control group,Daying-35 group,and Bushen Huayu Recipe group were 0.491±0.020,0.379±0.014,and 0.383±0.019,respectively,which was higher than the blank control group,and the difference was statistically significant(P<0.01);Compared with the model control group,the Da Ying-35 group and the Bushen Huayu Recipe group were lower than the model control group,and the difference was statistically significant(P<0.01).PR expression:Compared with the blank control group,the model control group and the Bushen Huayu Recipe group were 0.292±0.02 and 0.215±0.012,respectively,which were significantly higher than the blank control group,and the difference was statistically significant(P<0.01).The 35 groups were 0.210±0.014,which was higher than the blank control group,and the difference was statistically significant(P<0.05).Compared with that the model control group,the Daying-35 group and the Bushen Huayu recipe group were significantly lower than the model control group(P<0.05).Conclusion:Bushen Huayu Recipe can improve the expression of endometrial pinocytosis and decrease the levels of ER and PR protein in polycystic ovary model rats,which may be one of the mechanisms to improve endometrial receptivity.
引文
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[10]李燕,张庆文,王红彬.脱氢表雄酮诱导SD大鼠多囊卵巢综合征模型的实验研究[J].中国医药科学,2013,4(7):35-37.
[11]张晓薇,邝健全,曾爱群,等.脱氢表雄酮诱导多囊卵巢综合征动物模型的研究[J].广州医学院学报,2000,30(3):14-17.
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[13]贺石林,王健,王净净.中医科研设计与统计学[M].长沙:湖南科学技术出版社,2007:48.
[14]AGHAJANOVA L,STAVREUS EVERS A,NIKAS Y,et al.Co-expression of pinopodes and leukemia inhibitory factor,as well as its receptor,in hunman endometrium[J].Fertil Steril,2003,79(3):808-814.
[15]STEPHANIE A,FISHER M,ROBERT L,et al.A randomized double-blind comparison of the effects of clomiphene citrate and thear-omatase inhibitor letrozole on ovulatory function in normal women[J].Fertil Steril,2002,78(2):280-285.
[16]单书健,陈子华.古今名医临证金鉴妇科卷(下)[M].北京:中国中医药出版社,1998:288.
[17]陈莹,吴玥,宋金春.补骨脂化学成分及生物活性研究进展[J].实用药物与临床,2016,21(9):1184-1188.
[2]MITRA S,NAYAK P K,AGRAWAL S.Laparoscopic ovarian drilling:An alternative but not the ultimate in the management of polycystic ovary syndrome[J].Nat Sci Biol Med,2015,6(1):40-48.
[3]VITEK W,ALUR S,HOEGER K M.Off-label drug use in the treatment of polycystic ovary syndrome[J].Fertil Steril,2015,103(3):605-611.
[4]谢幸,苟文丽.妇产科学[M].北京:人民卫生出版社,2013:361.
[5]乐杰.妇产科学[M].北京:人民卫生出版社,2008:313-316.
[6]BALEN A.Polycystic ovary syndrome and cancer[J].Hum Keprod Update,2011,7(5):522-525.
[7]YOUNIS A,HAWKINS K,DIZER S,et al.Ovarian stimulation for IUI-IVF alter1 cytokine,chemokine,and antioxidant levels in women with endometriosis,PCOS,or unexplained infertility[J].Ferti Steri,2013,25(14):1003-1004.
[8]姚绚.补肾化瘀方对PCOS患者孕激素受体表达的影响[D].长沙:湖南中医药大学,2013.
[9]余曦明.补肾化瘀方对PCO大鼠卵巢颗粒细胞上ER和AR基因表达的影响[D].长沙:湖南中医药大学,2011.
[10]李燕,张庆文,王红彬.脱氢表雄酮诱导SD大鼠多囊卵巢综合征模型的实验研究[J].中国医药科学,2013,4(7):35-37.
[11]张晓薇,邝健全,曾爱群,等.脱氢表雄酮诱导多囊卵巢综合征动物模型的研究[J].广州医学院学报,2000,30(3):14-17.
[12]徐叔云,卞如镰,陈修.药理实验方法学[M].北京:人民卫生出版社,2006:1861.
[13]贺石林,王健,王净净.中医科研设计与统计学[M].长沙:湖南科学技术出版社,2007:48.
[14]AGHAJANOVA L,STAVREUS EVERS A,NIKAS Y,et al.Co-expression of pinopodes and leukemia inhibitory factor,as well as its receptor,in hunman endometrium[J].Fertil Steril,2003,79(3):808-814.
[15]STEPHANIE A,FISHER M,ROBERT L,et al.A randomized double-blind comparison of the effects of clomiphene citrate and thear-omatase inhibitor letrozole on ovulatory function in normal women[J].Fertil Steril,2002,78(2):280-285.
[16]单书健,陈子华.古今名医临证金鉴妇科卷(下)[M].北京:中国中医药出版社,1998:288.
[17]陈莹,吴玥,宋金春.补骨脂化学成分及生物活性研究进展[J].实用药物与临床,2016,21(9):1184-1188.