摘要
目的基于Ang/Tie-2信号通路,探讨补肾活血方孕前导法给药调控肾虚血瘀-胚胎着床障碍病证,结合模型大鼠种植窗期子宫组织血管生成的助孕机制。方法阴道涂片HE染色法观察,选取动情周期规律的健康未交配雌性SD大鼠60只,随机分为5组:空白组、模型组、阿司匹林组、补肾活血方高剂量组、补肾活血方低剂量组,动情前期时入组,分组给药并合笼,于妊娠第5天处死,腹主动脉采血后,迅速剖取子宫并制片,采用光镜观察各组大鼠子宫内膜组织形态,采用免疫组化法(IHC)检测各组大鼠子宫组织中VEGF、FLK-1表达水平及螺旋动脉数量,采用Western-Blot检测各组大鼠子宫组织中Ang-1、Ang-2、Tie-2蛋白表达水平,采用Real-Time PCR检测各组大鼠子宫组织中Ang-1、Ang-2、Tie-2 mRNA表达水平。结果给药组能明显改善肾虚血瘀-胚胎着床障碍病证结合模型大鼠种植窗期子宫内膜组织形态,并接近空白组水平;模型组大鼠子宫湿重、子宫脏器指数、螺旋动脉数量、VEGF、FLK-1及子宫组织Tie-2蛋白表达、Ang-1mRNA、Ang-2mRNA和Tie-2mRNA表达等均较空白组降低(P<0.05);补肾活血方高剂量组大鼠妊娠第5天子宫组织VEGF、FLK-1、螺旋动脉数量、Ang-1蛋白、Tie-2蛋白及Ang-1mRNA、Tie-2mRNA表达量较模型组明显增加(P<0.01)。结论补肾活血方孕前导法给药,改善肾虚血瘀-胚胎着床障碍病证结合模型大鼠种植窗期子宫内膜组织形态,促进子宫组织血管生成,改善组织血瘀状态,促进胚胎着床,可能与其提高大鼠子宫组织VEGF、FLK-1、Ang-1、Tie-2表达水平,并维持Ang-2正常表达有关。
Objective To investigate the effect on the pre-pregnancy administration with Bushen Huoxue Recipe enema with the influence on the expression of uterus tissue Ang-1,Ang-2,Tie-2,VEGF and FLK-1 during implantation window of rats with kidney-deficiency and blood stasis-embryo implantation dysfunction based on Ang/Tie-2 signaling pathway to interpretate the mechanism on assisting pregnancy.Methods The vaginal smear was observed by HE staining of sixty healthy female unconjugated SD rats with estrous cycle were randomly divided into 5 groups:blank group,model group,control group,Bushen Huoxue Recipe high-dose group and Bushen Huoxue Recipe low-dose group.All rats were enrolled in the early stage of estrus,grouped and caged with drug administration,and operation on the fifth day of pregnancy.The uterus was quickly dissected and prepared after blood collection from the abdominal aorta.Morphological structure of the endometrial tissues of each group were observed by light microscopy.Immunohistochemistry(IHC)was used to detect the expression level of VEGF,FLK-1 and spiral arteries count in the uterus of each group.Western-Blot test was used to analysis the protein expression level of Ang-1,Ang-2 and Tie-2 in the uterus tissues of each group.Real-Time PCR test was used to analysis the expression level of Ang-1,Ang-2 and Tie-2 mRNA in uterus of each group.Results The endometrial tissue cells in the blank group and the drug-treated group showed superiority in morphology and arrangement compared with the model group.Compared with the blank group,the model group uterine wet weight,uterine organs coefficient,spiral arteries count,VEGF,FLK-1 and uterine tissue Tie-2 protein expression,Ang-1 mRNA,Ang-2 mRNA and Tie-2 mRNA expression were significantly decreased(P<0.05).Compared with the model group,the expression level of VEGF,FLK-1,spiral arteries count,Ang-1 protein,Tie-2 protein,Ang-1 mRNA and Tie-2 mRNA in the uterus tissue of rats from the high-dose group with Bushen Huoxue Recipe was significantly increased.The difference wa
作者
严莉
郭珮
周航
夏宛廷
黄金珠
曾鹏飞
曾倩
Yan Li;Guo Pei;Zhou Hang;Xia Wanting;Huang Jinzhu;Zeng Pengfei;Zeng Qian(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第6期1862-1870,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81973901):补肾活血导法调控LXA4信号轴重塑子宫内膜种植窗期“炎症-免疫稳态”的助孕机制研究,负责人:曾倩
成都市科学技术局科技惠民技术研发项目(2016-HM01-00343-SF):补肾活血方直肠导入调控肾虚血瘀不孕大鼠子宫内膜LXA4表达水平的助孕机制研究,负责人:曾倩。
