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

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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
- Published：05 March 2023，
  
  Published Online：06 April 2022，
  
  Received：07 January 2022，
- 稿件说明：
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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

雷公藤多苷片菟丝子总黄酮配伍卵巢生殖干细胞神经源性位点缺口同源蛋白（Notch）信号通路

Keywords

Tripterygium wilfordii polyglycoside tablets（TWPT）Cuscutae Semen total flavonoidscombinationovarian germline stem cellsneurogenic locus notch homolog protein （Notch） signaling pathway

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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