菟丝子总黄酮对卵巢早衰大鼠卵巢功能的作用研究
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摘要
目的研究菟丝子总黄酮(TFSC)对卵巢早衰大鼠卵巢功能的作用。方法使用一次性环磷酰胺腹腔注射联合皮内注射白消安法制造大鼠卵巢早衰模型。提取与分离TFSC。将90只雌性健康SD大鼠采用随机数字法分为正常组、模型组、雌激素对照组以及TFSC低、中、高剂量(35、70、140 mg/kg)组,每组15只,除正常组外其余均造模,造模成功后开始给药;观察大鼠动情周期,给药10 d后处死大鼠,称重,取血清检测大鼠的卵泡刺激素、雌二醇水平;取大鼠的卵巢组织,称重,计算大鼠卵巢指数; 10%甲醛溶液固定后制片,苏木精-伊红染色法染色,显微镜下观察卵巢的形态学变化;观察卵泡发育,计数卵泡数量。结果模型组表现为持续的动情间期、无动情前期或动情期。正常组、雌激素对照组及TFSC低、中、高剂量组的动情期分别为4～5 d、5～6 d、7～8 d、6～7 d、5～6 d。各组大鼠的雌激素、卵泡刺激素水平均不同程度的降低(P <0. 05)。与正常组比较,模型组、雌激素对照组及TFSC低、中、高剂量组的体重、卵巢指数均降低(P <0. 05),但与模型组比较,TFSC高剂量组大鼠体重、卵巢指数均升高(P <0. 05)。模型组、雌激素对照组及TFSC低、中、高剂量组总卵泡数少于正常组,雌激素对照组及TFSC中、高剂量组多于模型组(P <0. 05);模型组、雌激素对照组及TFSC低、中剂量组生长卵泡少于正常组,雌激素对照组及TFSC中、高剂量组多于模型组(P <0. 05)。病理形态学观察模型组卵巢明显萎缩,生长卵泡数量减少。结论TFSC对卵巢早衰大鼠的卵巢功能有明显的恢复作用,可增加卵巢早衰大鼠的卵巢重量及卵泡数量,提高雌激素水平,对卵巢早衰有明显疗效。
Objective To study the effect of total flavonoids from semen cuscutae( TFSC) on rat model with premature ovarian failure. Methods Ninety rats were randomly divided into six groups: normal group,model group,the estrogen control group,and the TFSC group( high-140 mg,medium-70 mg,and low-35 mg dose groups),15 each group. Except the blank group,all animals were modeled( established by one time intraperitoneal injection of cyclophosphamide combined with intradermal injection of busulfan). We observed the estrous cycle in rats,and after 10 days of administration,the rats were sacrificed and weighed. Serum was taken to detect the levels of follicle stimulating hormone and estradiol in rats; ovarian tissue was taken to weigh and calculate the ovarian index of rats; 10% formaldehyde solution fixation was done to make slices,and hematoxylin-eosin staining was used to observe the morphological changes of ovaries under the microscope; follicle development was observed and the number of follicles was counted. Results The model group showed continuous interestrus,no preestrus or estrus. The estrus period of the normal group,estrogen control group and low-dose,medium-dose and high-dose TFSC group was 4-5 d,5-6 d,7-8 d,6-7 d,5-6 d,respectively. The levels of estrogen and follicular stimulating hormone in each group were decreased in different degrees( P < 0. 05). Compared with the normal group,the body weight and ovarian index of the model group,estrogen control group and TFSC groups were lower( P < 0. 05),but compared with the model group,the body weight and ovarian index of the high dose TFSC group were higher( P < 0. 05). The total numbers of follicles in the model group,estrogen control group and low,medium and high dose TFSC groups were lower than that in the normal group,while in the estrogen control group and medium and high dose TFSC group were more than that in the model group( P < 0. 05). The growing follicles in the model group,estrogen control group and low and medium dose TFSC group were less than those in the normal group,and in the estrogen control group and medium and high dose TFSC group were more than that in the model group( P < 0. 05). As for the pathomorphological observation,the ovaries were significantly atrophied and the number of growing follicles decreased in the model group. Conclusion TFSC can obviously restore the ovarian function of the rats with premature ovarian failure,increase the ovarian volume and weight of the rats with premature ovarian failure and the number of follicles,and improve the level of estrogen,showing obvious curative effect on premature ovarian failure.
Objective To study the effect of total flavonoids from semen cuscutae( TFSC) on rat model with premature ovarian failure. Methods Ninety rats were randomly divided into six groups: normal group,model group,the estrogen control group,and the TFSC group( high-140 mg,medium-70 mg,and low-35 mg dose groups),15 each group. Except the blank group,all animals were modeled( established by one time intraperitoneal injection of cyclophosphamide combined with intradermal injection of busulfan). We observed the estrous cycle in rats,and after 10 days of administration,the rats were sacrificed and weighed. Serum was taken to detect the levels of follicle stimulating hormone and estradiol in rats; ovarian tissue was taken to weigh and calculate the ovarian index of rats; 10% formaldehyde solution fixation was done to make slices,and hematoxylin-eosin staining was used to observe the morphological changes of ovaries under the microscope; follicle development was observed and the number of follicles was counted. Results The model group showed continuous interestrus,no preestrus or estrus. The estrus period of the normal group,estrogen control group and low-dose,medium-dose and high-dose TFSC group was 4-5 d,5-6 d,7-8 d,6-7 d,5-6 d,respectively. The levels of estrogen and follicular stimulating hormone in each group were decreased in different degrees( P < 0. 05). Compared with the normal group,the body weight and ovarian index of the model group,estrogen control group and TFSC groups were lower( P < 0. 05),but compared with the model group,the body weight and ovarian index of the high dose TFSC group were higher( P < 0. 05). The total numbers of follicles in the model group,estrogen control group and low,medium and high dose TFSC groups were lower than that in the normal group,while in the estrogen control group and medium and high dose TFSC group were more than that in the model group( P < 0. 05). The growing follicles in the model group,estrogen control group and low and medium dose TFSC group were less than those in the normal group,and in the estrogen control group and medium and high dose TFSC group were more than that in the model group( P < 0. 05). As for the pathomorphological observation,the ovaries were significantly atrophied and the number of growing follicles decreased in the model group. Conclusion TFSC can obviously restore the ovarian function of the rats with premature ovarian failure,increase the ovarian volume and weight of the rats with premature ovarian failure and the number of follicles,and improve the level of estrogen,showing obvious curative effect on premature ovarian failure.
引文
[1]李茜西,吕德欣.卵巢早衰的病因及治疗研究进展[J].中国医药导报,2014,11(30):164-168.
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[3]徐苓,宋亦军.卵巢早衰的临床表现和诊断标准[J].实用妇产科杂志,2003,19(4):195-196.
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[5]沈炜,陈学奇.调经汤对卵巢早衰模型大鼠卵巢功能影响的实验研究[J].浙江中西医结合杂志,2017,27(12):1040-1043.
[6]潘文灏,许志超,赵余庆.菟丝子的生物活性与临床应用研究进展[J].亚太传统医药,2008,4(4):47-51.
[7]刘新玉,刘昱磊,罗颂平.菟丝子提取物含药血清对人早孕滋养层细胞增殖及凋亡的影响[J].中国生化药物杂志,2016,36(5):43-46,50.
[8]任献青,郑贵珍,苏杭,等.菟丝子黄酮对雷公藤多苷片致生精细胞周期阻滞、凋亡及相关蛋白表达降低的影响[J].药物评价研究,2018,41(1):55-60.
[9]胡道敏.加减苁蓉菟丝子丸治疗卵巢早衰疗效分析[J].西部中医药,2010,23(9):43-44.
[10]赵素霞,刘会丽,江红.菟丝子黄酮通过调节Wnt/β-catenin信号通路干预去卵巢大鼠骨代谢的机制研究[J].临床和实验医学杂志,2018,17(1):25-28.
[11]柯江维,王建红,赵宏.菟丝子黄酮对心理应激雌性大鼠海马-下丘脑-垂体-卵巢轴性激素受体的影响[J].中草药,2006,37(1):90-92.
[12]王晓红.抗衰复巢汤与激素补充疗法结合治疗卵巢早衰的临床观察[J].中国现代药物应用,2015,9(20):134-136.
[13]刘慧莹,任春娥,姜爱芳.两种造模方法导致大鼠卵巢早衰模型的比较研究[J].中国妇幼保健,2015,28(30):4869-4871.
[14]马灵芝,金季玲,夏天.补肾调冲方对半乳糖致卵巢早衰大鼠卵巢功能的影响[J].江苏中医药,2011,43(6):91-92.
[15]李靖,李炫诚,吴云霞,等.确定小鼠动情周期的三种方法[J].实验动物科学,2007,27(1):67-68.
[16]李晓红,闫宏,刘绪红,等.补肾养血方对卵巢早衰小鼠动情周期及卵巢指数的影响[J].解剖学研究,2014,36(3):204-207.
[17]蔡华普,徐晶,孟庆玲.卵巢功能早衰中医药治疗近况[J].山东中医杂志,2004,23(9):574-575.
[18]张立.卵巢早衰36例分析及其治疗[J].中医药学刊,2003,21(4):619.
[19]王建国,张会臣.菟丝子治疗肾虚型男性不育症19例[J].河北中医,2001,23(1):53.
[20]刘小红,霍君怡,罗前,等.吴克明教授运用新加苁蓉菟丝子丸治疗卵巢早衰助孕成功经验撷粹[J].云南中医中药杂志,2016,37(4):19-22.
[21]李炳如,佘运初.补肾中药对下丘脑-垂体-性腺轴功能的影响[J].中医杂志,1984,25(7):63.
[22]王建红,王敏璋,欧阳栋,等.菟丝子黄酮对心理应激雌性大鼠下丘脑β-EP与腺垂体FSH、LH的影响[J].中药材,2002,25(12):886-888.
[23]王建红,王敏璋,伍庆华,等.菟丝子黄酮对应激大鼠卵巢内分泌的影响[J].中草药,2002,33(12):1099-1101.
[2]丰有吉,沈悭.妇产科学[M].北京:人民卫生出版社,2010:252.
[3]徐苓,宋亦军.卵巢早衰的临床表现和诊断标准[J].实用妇产科杂志,2003,19(4):195-196.
[4]Jankowska K.Premature ovarian failure[J].Menopausal Review,2017,16(2):51-56.
[5]沈炜,陈学奇.调经汤对卵巢早衰模型大鼠卵巢功能影响的实验研究[J].浙江中西医结合杂志,2017,27(12):1040-1043.
[6]潘文灏,许志超,赵余庆.菟丝子的生物活性与临床应用研究进展[J].亚太传统医药,2008,4(4):47-51.
[7]刘新玉,刘昱磊,罗颂平.菟丝子提取物含药血清对人早孕滋养层细胞增殖及凋亡的影响[J].中国生化药物杂志,2016,36(5):43-46,50.
[8]任献青,郑贵珍,苏杭,等.菟丝子黄酮对雷公藤多苷片致生精细胞周期阻滞、凋亡及相关蛋白表达降低的影响[J].药物评价研究,2018,41(1):55-60.
[9]胡道敏.加减苁蓉菟丝子丸治疗卵巢早衰疗效分析[J].西部中医药,2010,23(9):43-44.
[10]赵素霞,刘会丽,江红.菟丝子黄酮通过调节Wnt/β-catenin信号通路干预去卵巢大鼠骨代谢的机制研究[J].临床和实验医学杂志,2018,17(1):25-28.
[11]柯江维,王建红,赵宏.菟丝子黄酮对心理应激雌性大鼠海马-下丘脑-垂体-卵巢轴性激素受体的影响[J].中草药,2006,37(1):90-92.
[12]王晓红.抗衰复巢汤与激素补充疗法结合治疗卵巢早衰的临床观察[J].中国现代药物应用,2015,9(20):134-136.
[13]刘慧莹,任春娥,姜爱芳.两种造模方法导致大鼠卵巢早衰模型的比较研究[J].中国妇幼保健,2015,28(30):4869-4871.
[14]马灵芝,金季玲,夏天.补肾调冲方对半乳糖致卵巢早衰大鼠卵巢功能的影响[J].江苏中医药,2011,43(6):91-92.
[15]李靖,李炫诚,吴云霞,等.确定小鼠动情周期的三种方法[J].实验动物科学,2007,27(1):67-68.
[16]李晓红,闫宏,刘绪红,等.补肾养血方对卵巢早衰小鼠动情周期及卵巢指数的影响[J].解剖学研究,2014,36(3):204-207.
[17]蔡华普,徐晶,孟庆玲.卵巢功能早衰中医药治疗近况[J].山东中医杂志,2004,23(9):574-575.
[18]张立.卵巢早衰36例分析及其治疗[J].中医药学刊,2003,21(4):619.
[19]王建国,张会臣.菟丝子治疗肾虚型男性不育症19例[J].河北中医,2001,23(1):53.
[20]刘小红,霍君怡,罗前,等.吴克明教授运用新加苁蓉菟丝子丸治疗卵巢早衰助孕成功经验撷粹[J].云南中医中药杂志,2016,37(4):19-22.
[21]李炳如,佘运初.补肾中药对下丘脑-垂体-性腺轴功能的影响[J].中医杂志,1984,25(7):63.
[22]王建红,王敏璋,欧阳栋,等.菟丝子黄酮对心理应激雌性大鼠下丘脑β-EP与腺垂体FSH、LH的影响[J].中药材,2002,25(12):886-888.
[23]王建红,王敏璋,伍庆华,等.菟丝子黄酮对应激大鼠卵巢内分泌的影响[J].中草药,2002,33(12):1099-1101.