Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with <italic style="font-style: italic">Tripterygium wilfordii</italic> Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway

YANG Yibo; XU Ying; WANG Lu; ZOU Zhao; XU Panyu; DU Hanqian; LI Jiashan; LIN Na

doi:10.13422/j.cnki.syfjx.20220944

您当前的位置：

首页 >

文章列表页 >

Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway

更新时间：2023-02-07

- Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 5, Pages: 9-15(2023)
- 作者机构：
  
  中国中医科学院中药研究所，北京 100700
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20220944
  CLC： R2-0;R33;R289;R593.22
- Received：07 January 2022，
  
  Published Online：06 April 2022，
  
  Published：05 March 2023
- 稿件说明：
移动端阅览
杨一博,徐颖,王璐等.基于Notch信号通路探索菟丝子总黄酮配伍雷公藤多苷对生理小鼠卵巢生殖干细胞的作用及机制[J].中国实验方剂学杂志,2023,29(05):9-15.

YANG Yibo,XU Ying,WANG Lu,et al.Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):9-15.
杨一博,徐颖,王璐等.基于Notch信号通路探索菟丝子总黄酮配伍雷公藤多苷对生理小鼠卵巢生殖干细胞的作用及机制[J].中国实验方剂学杂志,2023,29(05):9-15. DOI： 10.13422/j.cnki.syfjx.20220944.

YANG Yibo,XU Ying,WANG Lu,et al.Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):9-15. DOI： 10.13422/j.cnki.syfjx.20220944.

摘要

目的

探讨菟丝子总黄酮配伍雷公藤多苷片对雌性生理小鼠卵巢生殖干细胞的作用，并从神经源性位点缺口同源蛋白（Notch）信号通路探讨其作用机制。

方法

将60只5周龄雌性昆明种小鼠随机分为正常组，1、2倍临床等效剂量雷公藤多苷片组（低、高剂量组13.65、27.3 mg·kg

-1

·d

-1

），菟丝子总黄酮（低、高剂量组150、300 mg·kg

-1

·d

-1

），配伍组（雷公藤多苷片低剂量+菟丝子总黄酮低剂量组），每组10只，连续给药3周后取材，计算子宫、卵巢脏脑指数，光镜下观察卵巢组织病理形态学改变；酶联免疫吸附测定法（ELISA）检测血清中雌二醇含量；免疫荧光法观察卵巢上皮的卵巢生殖干细胞标志物果蝇VASA基因同源类似物（Mvh）、八聚体结合转录因子4（Oct4）、酪氨酸激酶受体（c-kit）、同源框蛋白质（Nanog）及Notch信号通路分子神经源性基因Notch同源蛋白1（Notch1）、Hes家族BHLH转录因子1（Hes1）、锯齿典型Notch配体1（Jagged1）的表达。

结果

与正常组比较，雷公藤多苷片低剂量组和高剂量组对小鼠子宫、卵巢的脏脑系数及形态学差异无统计学意义，仅可致小鼠闭锁卵泡数显著增加（

0.01）；雷公藤多苷片低剂量对卵巢生殖干细胞标志物和Notch信号通路分子的表达有降低趋势，雷公藤多苷片高剂量下Mvh、c-kit、Nanog的表达水平明显降低（

0.05，

0.01），Notch1、Hes1表达水平显著降低（

0.01）；菟丝子总黄酮低剂量组上述指标均明显升高（

0.05，

0.01）；与雷公藤多苷片低剂量组比较，配伍组闭锁卵泡数显著降低（

0.01），Mvh、Nanog表达水平显著升高（

0.01），Notch1、Hes1表达水平明显升高（

0.05，

0.01）。

结论

卵巢生殖干细胞是雷公藤多苷片生殖毒性的源头靶点，菟丝子总黄酮可参与调控卵巢生殖干细胞途径缓解雷公藤多苷片所致的生殖损伤，其作用机制可能与Notch信号通路相关。

Abstract

Objective

To observe the effect of Cuscutae Semen total flavonoids combined with

Tripterygium wilfordii

polyglycoside tablets （TWPT） on ovarian germline stem cells of female physiological mice through neurogenic locus notch homolog （Notch） signaling pathway.

Method

Sixty female Kunming mice （5 weeks old） were randomly divided into normal group， Tripterygium wilfordii polyglycoside tablets low-， high-dose groups （13.65 mg·kg

-1

·d

-1

and 27.3 mg·kg

-1

·d

-1

，

1 and 2 times clinical equivalent dose）， Cuscutae Semen total flavonoids low- and high-dose groups （150 mg·kg

-1

·d

-1

and 300 mg·kg

-1

·d

-1

）， and combination group （13.65 mg·kg

-1

·d

-1

TWPT and 150 mg·kg

-1

·d

-1

Cuscutae Semen total flavonoids）， with 10 in each group. After 3 weeks of continuous administration， the uterus/brain and ovarian/brain indexes were calculated， and the pathological changes of ovarian tissue were observed under light microscope. The content of estradiol in serum was determined by enzyme linked immunosorbent assay （ELISA）. Immunofluorescence assay was performed to observe the expressions of germline stem cell markers in ovarian epithelium， including mouse vasa homologue （Mvh）， octamer-binding transcription factor 4 （Oct4）， tyrosine-protein kinase receptor （c-kit）， Nanog， Notch signaling pathway molecules， neurogenic locus notch homolog protein 1 （Notch1）， hes family BHLH transcription factor 1（Hes1）， and jagged canonical Notch ligand 1 （JAG1）.

Result

Compared with the normal group， low and high doses of TWPT had no significant effect on the uterus/brain and ovary/brain indexes and the uterus and ovary morphologies of mice， while only the number of atretic follicles was increased （

0.01）. The expressions of ovarian germline stem cell markers and Notch signaling pathway molecules had a decreasing trend in TWPT low-dose group， while the expressions of Mvh， c-kit， and Nanog were down-regulated （

0.05，

0.01） and the expressions of Notch1 and Hes1 were also reduced （

0.01） in TWPT high-dose group. However， the above indexes were increased in Cuscutae Semen total flavonoids low-dose group （

0.05，

0.01）. Compared with the low does of TWPT group， the combination group had a decrease in the increased number of atretic follicles （

0.01）， an improvement in the down-regulated expressions of Mvh and Nanog （

0.01）， and an increase in the expressions of Notch1 and Hes1 （

0.05，

0.01）.

Conclusion

Ovarian germline stem cells are the source target of the reproductive toxicity of TWPT. Cuscutae Semen total flavonoids participate in the regulation of the germline stem cell pathways to alleviate the reproductive toxicity caused by TWPT， and its mechanism of action may be related to the Notch signaling pathway.

关键词

Keywords

references

徐颖，樊媛芳，赵元，等 . 近40年雷公藤生殖毒性研究概述［J］. 中国中药杂志， 2019 ， 44 （ 16 ）： 3406 - 3414 .

崔瑞琴，丁樱 . 菟丝子总黄酮对雷公藤多苷片所致生殖损伤雌鼠卵巢损伤表达的影响［J］. 辽宁中医药大学学报， 2009 ， 11 （ 8 ）： 246 - 247 .

JOHNSON J ， CANNING J ， KANEKO T ， et al . Germline stem cells and follicular renewal in the postnatal mammalian ovary ［J］. Nature ， 2004 ， 428 （ 6979 ）： 145 - 150 .

TANG Z ， WEI J ， YU Y ， et al . Gamma-secretase inhibitor reverts the Notch signaling attenuation of osteogenic differentiation in aged bone marrow mesenchymal stem cells ［J］. Cell Biol Int ， 2016 ， 40 （ 4 ）： 439 - 447 .

ZYGMUNT D A ， SINGHAL N ， KIM M L ， et al . Deletion of pofut1 in mouse skeletal myofibers induces muscle aging-related phenotypes in cis and in trans ［J］. Mol Cell Biol ， 2017 ， doi： 10.1128/MCB.00426-16 http://dx.doi.org/10.1128/MCB.00426-16 .

MANOKAWINCHOKE J ， NATTASIT P ， THONGNGAM T ， et al . Indirect immobilized Jagged1 suppresses cell cycle progression and induces odonto/osteogenic differentiation in human dental pulp cells ［J］. Sci Rep ， 2017 ， 7 （ 1 ）： 10124 .

苗明三，彭孟凡，闫晓丽，等 . 菟丝子总黄酮对来曲唑致多囊卵巢综合征大鼠模型的影响［J］. 中国实验方剂学杂志， 2019 ， 25 （ 16 ）： 17 - 23 .

陶玲，白敦耀，臧振中，等 . 雷公藤甲素阿魏酸醇质体凝胶经皮给药的长期毒性［J］. 中国实验方剂学杂志， 2022 ， 28 （ 7 ）： 95 - 104 .

樊媛芳，徐颖，苏晓慧，等 . 雷公藤多苷片片对Ⅱ型胶原诱导性关节炎雌性大鼠生殖毒性的影响［J］. 中国中药杂志， 2019 ， 44 （ 16 ）： 3486 - 3493 .

YANG H ， YAO X ， TANG F ， et al . Characterization of female germline stem cells from adult mouse ovaries and the role ofrapamycin on them ［J］. Cytotechnology ， 2018 ， 70 （ 2 ）： 843 - 854 .

WANG N ， SATIRAPOD C ， OHGUCHI Y ， et al . Genetic studies in mice directly link oocytes produced during adulthood to ovarian function and natural fertility ［J］. Sci Rep ， 2017 ， 7 （ 1 ）： 10011 .

CELIK O ， CELIK E ， TURKCUOGLU I ， et al . Germline cells in ovarian surface epithelium of mammalians：A promising notion ［J］. Reprod Biol Endocrinol ， 2012 ， 10 ： 112 .

WHITE Y A ， WOODS D C ， TAKAI Y ， et al . Oocyte formation by mitotically active germ cells purified from ovaries of reproductiveage women ［J］. Nat Med ， 2012 ， 18 （ 3 ）： 413 - 421 .

PATEL H ， BHARTIYA D ， PARTE S . Further characterization of adult sheep ovarian stem cells and their involvement in neo-oogenesis and follicle assembly ［J］. J Ovarian Res ， 2018 ， 11 （ 1 ）： 3 .

WOODS D C ， WHITE Y A ， TILLY J L . Purification of oogonial stem cells from adult mouse and human ovaries： An assessment of the literature and a view toward the future ［J］. Repord Sci ， 2013 ， 20 （ 1 ）： 7 - 15 .

SONG X ， CALL G B ， KIRILLY D ， et al . Notch signaling controls germline stem cell niche formation in the drosophila ovary ［J］. Development ， 2007 ， 134 （ 6 ）： 1071 - 1080 .

CHEN C L ， FU X F ， WANG L Q ， et al . Primordial follicle assembly was regulated by Notch signaling pathway in the mice ［J］. Mol Biol Rep ， 2014 ， 41 （ 3 ）： 1891 - 1899 .

NYSTUL T ， SPRADLING A . Regulation of epithelial stem cell replacement and follicle formation in the drosophila ovary ［J］. Genetics ， 2010 ， 184 （ 2 ）： 503 - 515 .

GALVÃO A M ， FERREIRA-DIAS G ， SKARZYNSKI D J . Cytokines and angiogenesis in the corpus luteum ［J］. Mediators Inflamm ， 2013 ： 420186 .

LIU J ， LU W G ， YE F ， et al . Hes1/Hes5 gene inhibitsdifferentiation via down-regulating Hash1 and promotes proliferation in cervical carcinoma cells ［J］. Int J Gynecol Cancer ， 2010 ， 20 （ 7 ）： 1109 - 1116 .

梁夏，潘泽政，叶海峰，等 . Notch信号通路参与卵巢生殖干细胞的调控及卵巢的衰老进程［J］. 中国生物化学与分子生物学报， 2019 ， 35 （ 4 ）： 428 - 435 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method

Mechanism of Yiqi Jiedu Prescriptions in Protection of Mitochondria in PC12 Cells Against Hypoxia Injury Based on Energy Metabolism

Prescription Compatibility Effect on Absorption of Major Components in Yanhusuo Extract

Related Author

LI Yuan

DU Hanqian

LI Jiashan

GUO Li

LI Zehui

LIN Na

Yu-hao DAI

Li-ming LIU

Related Institution

Institute of Chinese Materia Medica， Shanghai University of Traditional Chinese Medicine

Anhui University of Chinese Medicine

Key Laboratory of Resource and Compound Prescription of Traditional Chinese Medicine, Ministry of Education,Hubei University of Chinese Medicine

Sichuan Academy of Chinese Medicine Sciences

Department of Chinese Pharmacy,Beijing Jishuitan Hospital

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway

DOI：10.13422/j.cnki.syfjx.20220944

摘要

Abstract

关键词

Keywords

references