摘要
目的探讨益肾疏肝方促进胚胎着床的可能作用机制。方法雌、雄ICR小鼠按照3∶1的比例合笼饲养,次日清晨8:00检查雌鼠阴道栓,以见栓当日作为妊娠第1天。将见栓小鼠50只随机分为正常对照组、模型组、黄体酮组和中药低、高剂量组,每组10只。除正常对照组外其余各组小鼠妊娠第4天颈背部皮下注射米非司酮溶液(0.03 mg/kg)建立胚胎着床障碍模型。从妊娠第1天开始,模型组、正常对照组每日给予生理盐水0.2 ml/只灌胃,黄体酮组给予黄体酮溶液(浓度2 mg/ml)每日皮下注射20μl/只,中药低、高剂量组每日分别给予3.9 g/ml、15.6 g/ml的益肾疏肝方灌胃0.2 ml/只,连续给药7天。妊娠第8天处死小鼠,记录孕鼠数和着床总胚泡数,计算着床率和平均着床胚泡数;取下子宫,免疫组化法和Western blot法对子宫中MMP-2和MMP-9表达和蛋白含量进行检测。结果与正常对照组比较,模型组孕鼠数明显减少,着床率显著降低,平均着床胚泡数明显减少(P<0.05);与模型组比较,中药低、高剂量组和黄体酮组的平均着床胚泡数增高(P<0.05)。中药高剂量组、黄体酮组子宫中MMP-2的表达和蛋白含量较模型组显著升高(P<0.05);中药低、高剂量组和黄体酮组子宫中MMP-9的表达和蛋白含量明显高于模型组(P<0.05)。结论益肾疏肝方可以增加胚胎着床障碍小鼠的平均着床胚泡数,提高受孕率,可能与上调子宫中MMP-2和MMP-9的表达有关。
Objective To study the possible mechanism of Yishen Shugan Fang（ YSSGF） in promoting the embryo implantation. Methods Female and male ICR mice（ in proportion of 3： 1） were fed in the same cage. At8： 00 of the next morning,vaginal plug of the female mice were checked,and the day was regarded as the first day of pregnancy when saw the plug. Then 50 mice with plug were randomly divided into normal control group,model group,progesterone group,and the TCM low and high dose groups,with ten in each group. Except the normal control group,the other four groups were given subcutaneous injection with mifepristone solution（ 0. 03 mg / kg） into neck and back to establish a model of obstacle of embryo implantation on the fourth day of pregnancy. Starting from the first day of pregnancy,the model group and the normal control group were given by gavage with 0. 2 ml of normal saline every day,the progesterone group was given subcutaneous injection with progestin solution 20 μl（ 2 mg / ml） every day,the TCM low and high dose groups were given by gavage with 3. 9g / ml and 15. 6 g / ml of YSSGF 0. 2 ml,respectively every day,all for seven days. On the eighth day of pregnancy,the mice were sacrificed,and the number of pregnant mice and total blastocyst with implantation were recorded to calculate implantation rate and average number of blastocyst with implantation. The uteri were also removed to detect the expression of MMP-2 and MMP-9 and protein content by the immunohistochemical method and the Western blot method. Results Compared with the normal control group,the number of pregnant mice,the implantation rate and the average number of blastocyst with implantation decreased significantly in the model group（ P〈 0. 05）. Compared with the model group,the average number of blastocyst with implantation increased significantly in the TCM low and high dose groups and the progesterone group（ P 〈0. 05）,the expression of MMP-2 and protein content in uterus increased significantly in the TCM high dose group and the progesterone group（ P 〈0. 05）,and the expression of MMP-9 and protein content in uterus were significantly higher in the TCM low and high dose groups and the progesterone group（ P 〈0. 05）. Conclusion YSSGF may increase the average number of blastocyst with implantation of mice with obstacle of embryo implantation and improve the conception rate,and the underline mechanism may be related to increase the expression of MMP-2 and MMP-9 in uterus.
出处
《中医杂志》
CSCD
北大核心
2015年第18期1597-1600,共4页
Journal of Traditional Chinese Medicine
基金
国家中医药管理局重点专科(zk0901fk012)
上海市卫生局课题(ZYSNXD-CC-HPGC-JD-009)
关键词
胚胎着床障碍
益肾疏肝方
基质金属蛋白酶
obstacle of embryo implantation
Yishen Shugan Fang （益肾疏肝方）
matrix metalloproteinases
