中药活血补肾序贯治疗对子宫内膜异位症大鼠卵泡发育与颗粒细胞凋亡的影响
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摘要
目的:研究不同中药对子宫内膜异位症大鼠卵泡发育与卵巢颗粒细胞凋亡的影响。方法:采用异体子宫内膜移植法建立子宫内膜异位症大鼠模型,随机分为空白组、假手术组、模型组、活血消异方组、补肾助孕组、序贯组,于大鼠不同动情周期取材,计算性腺指数,HE染色计数各级卵泡,TUNEL染色比较各组不同动情周期间凋亡率的差异。结果:①窦前卵泡计数:模型组明显低于其余5组,与空白组、假手术组、补肾助孕组、序贯组比较,差异有统计学意义(P<0.05,P<0.01);②窦状卵泡计数:模型组显著低于其余5组(P<0.01);③4个动情周期模型组颗粒细胞凋亡率最高,动情前期显著高于其余5组(P<0.05,P<0.01)。结论:子宫内膜异位症影响卵泡发育,导致颗粒细胞凋亡,中药可提高子宫内膜异位症模型大鼠窦前卵泡与窦状卵泡数目,减少卵巢颗粒细胞凋亡,且于动情前期作用最显著。
Objective: To study the effects of different Chinese medicine on follicular development and ovarian granulosa cell apoptosis in rats with endometriosis. Methods: Endometriosis rat models were established by allogeneic endometrial transplantation. They were randomly divided into blank group, sham operation group, model group, Huoxue Xiaoyi Decoction group, Bushen Zhuyun Decoction group and sequential group. The gonadal index was calculated. Ovaries were counted by HE staining for the follicles at various levels. TUNEL staining to compare the differences in apoptotic rates between different groups of estrous cycles. Results: ①Preantral follicle counts: the model group was significantly lower than the other 5 groups,compared with the blank group, the sham operation group. Bushen Zhuyun Decoction group, and the Sequential treatment group, the difference was statistically significant(P<0.05, P<0.01). ②Antral follicle counts: the model group was significantly lower than the other 5 groups, the difference was statistically significant(P<0.01). ③Granulosa cell apoptosis rate, model group was the highest in the four estrous cycles, and was significantly higher than the other 5 models in proestrus, the difference was statistically significant(P<0.05, P<0.01). Conclusion: Endometriosis affects follicular development and leads to granulosa cell apoptosis. The number of preantral follicle and antral follicle are increased with the treatment of Chinese medicine. Ovarian granulosa cell apoptosis can be improved by treating with Chinese medicine, and the function of Chinese medicine has the most significant effect in proestrus.
Objective: To study the effects of different Chinese medicine on follicular development and ovarian granulosa cell apoptosis in rats with endometriosis. Methods: Endometriosis rat models were established by allogeneic endometrial transplantation. They were randomly divided into blank group, sham operation group, model group, Huoxue Xiaoyi Decoction group, Bushen Zhuyun Decoction group and sequential group. The gonadal index was calculated. Ovaries were counted by HE staining for the follicles at various levels. TUNEL staining to compare the differences in apoptotic rates between different groups of estrous cycles. Results: ①Preantral follicle counts: the model group was significantly lower than the other 5 groups,compared with the blank group, the sham operation group. Bushen Zhuyun Decoction group, and the Sequential treatment group, the difference was statistically significant(P<0.05, P<0.01). ②Antral follicle counts: the model group was significantly lower than the other 5 groups, the difference was statistically significant(P<0.01). ③Granulosa cell apoptosis rate, model group was the highest in the four estrous cycles, and was significantly higher than the other 5 models in proestrus, the difference was statistically significant(P<0.05, P<0.01). Conclusion: Endometriosis affects follicular development and leads to granulosa cell apoptosis. The number of preantral follicle and antral follicle are increased with the treatment of Chinese medicine. Ovarian granulosa cell apoptosis can be improved by treating with Chinese medicine, and the function of Chinese medicine has the most significant effect in proestrus.
引文
[1]张惜阴.实用妇产科学.2版.北京:人民卫生出版社,2003:764
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[3]Brosens I.Endometriosis and the outcome of in vitro fertilization.Fertility&Sterility,2004,81(5):1198-1200
[4]Olivennes F.Results of IVF in women with endometriosis.J Gynecd Obstet Bid Reprod,2003,32(2):45-47
[5]Hauzman E E,Garcia-Velasco J A,Pellicer A.Oocyte donation and endometriosis:What are the Lessons?Seminars in Reproductive Medicine,2013,31(2):173-177
[6]Assou S,Haouzi D,Vos J D,et al.Human cumulus cells as biomarkers for embryo and pregnancy outcomes.Molecular Human Reproduction,2010,16(8):531
[7]He Y,Deng H,Jiang Z,et al.Effects of melatonin on follicular atresia and granulosa cell apoptosis in the porcine.Molecular Reproduction&Development,2016,83(8):692-700
[8]Nakahara K,Saito H,Saito T,et al.Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program.Fertility&Sterility,1997,67(2):302
[9]Cora M C,Kooistra L,Travlos G.Vaginal cytology of the laboratory rat and mouse:Review and criteria for the staging of the estrous cycle using stained vaginal smears.Toxicologic Pathology,2015,43(6):776
[10]Guo,Junqi,Gao,et al.BMSCs reduce rat granulosa cell apoptosis induced by cisplatin and perimenopause.BMC Cell Biology,2013,14(1):18
[11]张泰魏,李燕,许露,等.逍遥散加减方对HPRL模型大鼠卵巢颗粒细胞凋亡及相关蛋白的影响.中华中医药杂志,2018,33(9):3909-3912
[12]Agarwal A,Saleh R A,Bedaiwy M A.Role of reactive oxygen species in the pathophysiology of human reproduction.Fertility&Sterility,2003,79(4):829-843
[13]Augoulea A,Creatsa M,Vrachnis N,et al.Pathogenesis of endometriosis:The role of genetics,inflammation and oxidative stress.Archives of Gynecology&Obstetrics,2012,286(1):99-103
[14]Ruder E H,Hartman T J,Blumberg J,et al.Oxidative stress and antioxidants:exposure and impact on female fertility.Human Reproduction Update,2008,14(4):345
[15]Jarkovska K,Martinkova J,Liskova L,et al.Proteome mining of human follicular fluid reveals a crucial role of complement cascade and key biological pathways in women undergoing in vitro fertilization.Journal of Proteome Research,2010,9(3):1289-1301
[16]Broi Michele Gomes Da,Jordao Alceu,Rui A Ferriani,et al.Oocyte oxidative DNA damage may be involved in minimal/mild endometriosis related infertility.Molecular Reproduction&Development,2017,85(2):128-136
[17]Shen M,Lin F,Zhang J,et al.Involvement of the up-regulated FoxO1 expression in follicular granulosa cell apoptosis induced by oxidative stress.Journal of Biological Chemistry,2012,287(31):25727-25740
[18]白俊,吴克明,郜然然.基于“肾主生殖”探赜细胞自噬机制与卵泡发育的微观联系.中国中药杂志,2018,43(5):921-925
[2]郎景和.重视疑难性子宫内膜异位症的临床与基础研究.中国实用妇科与产科杂志,2009,25(9):641-642
[3]Brosens I.Endometriosis and the outcome of in vitro fertilization.Fertility&Sterility,2004,81(5):1198-1200
[4]Olivennes F.Results of IVF in women with endometriosis.J Gynecd Obstet Bid Reprod,2003,32(2):45-47
[5]Hauzman E E,Garcia-Velasco J A,Pellicer A.Oocyte donation and endometriosis:What are the Lessons?Seminars in Reproductive Medicine,2013,31(2):173-177
[6]Assou S,Haouzi D,Vos J D,et al.Human cumulus cells as biomarkers for embryo and pregnancy outcomes.Molecular Human Reproduction,2010,16(8):531
[7]He Y,Deng H,Jiang Z,et al.Effects of melatonin on follicular atresia and granulosa cell apoptosis in the porcine.Molecular Reproduction&Development,2016,83(8):692-700
[8]Nakahara K,Saito H,Saito T,et al.Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program.Fertility&Sterility,1997,67(2):302
[9]Cora M C,Kooistra L,Travlos G.Vaginal cytology of the laboratory rat and mouse:Review and criteria for the staging of the estrous cycle using stained vaginal smears.Toxicologic Pathology,2015,43(6):776
[10]Guo,Junqi,Gao,et al.BMSCs reduce rat granulosa cell apoptosis induced by cisplatin and perimenopause.BMC Cell Biology,2013,14(1):18
[11]张泰魏,李燕,许露,等.逍遥散加减方对HPRL模型大鼠卵巢颗粒细胞凋亡及相关蛋白的影响.中华中医药杂志,2018,33(9):3909-3912
[12]Agarwal A,Saleh R A,Bedaiwy M A.Role of reactive oxygen species in the pathophysiology of human reproduction.Fertility&Sterility,2003,79(4):829-843
[13]Augoulea A,Creatsa M,Vrachnis N,et al.Pathogenesis of endometriosis:The role of genetics,inflammation and oxidative stress.Archives of Gynecology&Obstetrics,2012,286(1):99-103
[14]Ruder E H,Hartman T J,Blumberg J,et al.Oxidative stress and antioxidants:exposure and impact on female fertility.Human Reproduction Update,2008,14(4):345
[15]Jarkovska K,Martinkova J,Liskova L,et al.Proteome mining of human follicular fluid reveals a crucial role of complement cascade and key biological pathways in women undergoing in vitro fertilization.Journal of Proteome Research,2010,9(3):1289-1301
[16]Broi Michele Gomes Da,Jordao Alceu,Rui A Ferriani,et al.Oocyte oxidative DNA damage may be involved in minimal/mild endometriosis related infertility.Molecular Reproduction&Development,2017,85(2):128-136
[17]Shen M,Lin F,Zhang J,et al.Involvement of the up-regulated FoxO1 expression in follicular granulosa cell apoptosis induced by oxidative stress.Journal of Biological Chemistry,2012,287(31):25727-25740
[18]白俊,吴克明,郜然然.基于“肾主生殖”探赜细胞自噬机制与卵泡发育的微观联系.中国中药杂志,2018,43(5):921-925