主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2020, Vol. 43 ›› Issue (1): 35-42.doi: 10.3969/j.issn.1006-2157.2020.01.008

• 中药药理 • 上一篇    下一篇

补肾法对控制性超促排小鼠卵丘扩展分级及相关因子HAS2、TNFAIP6的影响*

梁潇1, 佟雪1, 杜惠兰1,2,3, 段彦苍1,2,3#   

  1. 1 河北中医学院中西医结合学院 河北 050091;
    2 河北省中西医结合生殖疾病协同创新中心;
    3 河北省中西医结合肝肾病证研究重点实验室
  • 收稿日期:2019-10-24 发布日期:2020-02-21
  • 通讯作者: 段彦苍,女,教授,博士生导师,主要研究方向:中西医结合妇科生殖内分泌,E-mail:duanyancang@163.com
  • 作者简介:梁潇,女,在读硕士生
  • 基金资助:
    *国家自然科学基金资助项目(No.81973893),国家大学生创新训练资助项目(No.201914432019),河北中医学院科研能力重点提升资助项目(No.KTZ 2019004)

Effects of kidney-invigorating method on the levels of cumulus expansion, HAS2 and TNFAIP6 in controlled ovarian hyperstimulation mice*

Liang Xiao1, Tong Xue1, Du huilan1,2,3, Duan Yancang1,2,3#   

  1. 1 College of Integrated Traditional Chinese and Western Medicine, Hebei University of Chinese Medicine, Hebei 050091, China;
    2 Hebei Collaborative Innovation Center of Integrated Chinese and Western Medicine on Reproductive Disease, Hebei 050091, China;
    3 Hebei Key Laboratory of Integrative Medicine on Liver-kidney Patterns, Hebei 050091, China
  • Received:2019-10-24 Published:2020-02-21
  • Contact: Prof. Duan Yancang, Ph. D., Doctoral Supervisor. College of Integrated Traditional Chinese and Western Medicine, Hebei University of Chinese Medicine. No.3, Xingyuan Road, Luquan District, Shijiazhuang city, Hebei Province 0500200, China. E-mail: duanyancang@163.com
  • Supported by:
    National Natural Science Foundation of China (No.81973893), National University students innovation training proiects (No.201914432019)

摘要: 目的 研究补肾调经Ⅱ号方、Ⅲ号方序贯给药对小鼠卵丘卵母细胞复合物(COCs)卵丘扩展分级情况及卵丘扩展相关因子透明质酸合成酶2(HAS2)、肿瘤坏死因子α刺激因子-6(TNFAIP6)表达的影响。 方法 将120小鼠随机分为正常组、模型组、补肾组。模型组、补肾组第1~9天腹腔注射醋酸丙氨瑞林(GnRHa),第9天同时注射血清促性腺激素(PMSG),48 h后注射绒毛膜促性腺激素(hCG),正常组注射等体积生理盐水;补肾组第1~9天灌胃补肾调经Ⅱ号方,第10~11天灌胃补肾调经Ⅲ号方,正常组与模型组灌胃等体积蒸馏水。注射生理盐水或hCG 6、9、12 h后在体式显微镜下取出卵丘卵母细胞复合物(COCs),观察卵丘扩展分级并计数,收集COCs,免疫荧光染色、实时荧光定量PCR检测卵丘扩展相关因子蛋白及mRNA的表达。结果 与正常组相比,模型组的卵丘扩展分级情况及卵丘扩展相关因子的mRNA表达水平差异无统计学意义(P>0.05);与正常组和模型组相比,补肾组的卵丘扩展分级呈现低级别扩展率降低、高级别扩展率升高的特点(P<0.05);注射生理盐水或hCG 6 h,正常组、模型组和补肾组3组间HAS2 mRNA的表达水平差异无统计学意义(P>0.05);注射生理盐水或hCG 9、12 h,与正常组和模型组相比,补肾组HAS2 mRNA的表达水平升高(P<0.05);注射生理盐水或hCG 6、9、12 h,补肾组TNFAIP6 mRNA的表达水平升高(P<0.05)。结论 补肾调经Ⅱ号方、Ⅲ号方序贯给药可促进小鼠COCs的卵丘扩展,推测其可能的机制与上调卵丘扩展相关因子HAS2、TNFAIP6 mRNA的表达有关。

关键词: 补肾调经方Ⅱ号方, 补肾调经Ⅲ号方, 卵丘卵母细胞复合物, 卵丘扩展, 透明质酸合成酶2, 肿瘤坏死因子α刺激因子-6, 小鼠

Abstract: Objective To study the effects of sequential administration of Bushen Tiaojing Fang II and III (Kidney-invigorating and Menstruation-regulating Decoction II and III, BSTJF II & BSTJF III) on cumulus-oocyte complex (COCs), the levels of cumulus expansion and related parameters of cumulus expansion (HAS2 and TNFAIP6) in mice. Methods 120 mice were randomly divided into normal group, model group and BSTJF group (n=40 in each group). From the 1st day to the 9th day, model group and BSTJF group were injected with GnRHa intraperitoneally, and on the 9th day, PMSG was injected simultaneously, and hCG was injected after 48 hours, while equal volume of normal saline was injected to the normal group. BSTJF group was given BSTJF II through gavage administration from the 1st day to the 9th day, and was given BSTJF III from the 10th day to the 11th day, while equal volume of distilled water was given to the normal group and the model group. The COCs were taken out under the microscope at 6 h, 9 h and 12 h after saline or hCG injection. The cumulus expansion was graded and counted, and COCs were collected. Protein expression of cumulus expansion related factors (HAS2 and TNFAIP6) was detected by immunofluorescence staining, and the mRNA expression of cumulus expansion related factors (HAS2 and TNFAIP6) was measured by using real-time fluorescent quantitative PCR. Results Compared with the normal group, there was no statistically significant difference in the level of cumulus expansion and the mRNA expression level of HAS2 and TNFAIP6 in the model group (P>0.05). Compared with the normal group and the model group, the cumulus expansion level of the BSTJF group showed a reduction of low-level expansion rate and an increase of high-level cumulus expansion rate (P<0.05). There was no statistically significant difference in the mRNA expression level of HAS2 among the normal group, model group and BSTTF group at 6 h after saline or hCG injection (P>0.05). The mRNA expression levels of HAS2 in the BSTJF group were higher than those in the normal group and the model group at 9 h and 12 h after saline or hCG injection (P<0.05). The mRNA expressions of TNFAIP6 in the BSTJF group at 6 h, 9 h, and 12 h after saline or hCG injection were higher than those of the normal group and the model group(P<0.05). Conclusion The sequential administration of Bushen Tiaojing Fang II and III could promote the cumulus expansion of COCs in mice, and it is speculated that the possible mechanism is related to the up-regulation of mRNA expression levels of cumulus expansion related factors HAS2 and TNFAIP6.

Key words: Bushen Tiaojing Fang II, Bushen Tiaojing Fang III, COCs, cumulus expansion, HAS2, TNFAIP6, mice

中图分类号: 

  • R285.5