Skip to content
2000
Volume 28, Issue 8
  • ISSN: 1386-2073
  • E-ISSN: 1875-5402

Abstract

Objective

This study aimed to investigate the possible mechanism through which acupuncture protects ovaries with Poor Ovarian Response (POR) in rats based on microRNA (miRNA).

Methods

Thirty-six SPF SD female non-pregnant rats aged 8 weeks were randomly divided into the blank group, model group, and acupuncture group, with 12 rats in each group. According to the group, the rats were given gavage of Tripterygium wilfordii polyglycosides suspension for 14 days to establish the model of POR, and then the rats were treated with acupuncture for 2 weeks, once a day, for 20 minutes.

Afterward, their hormone levels were measured, and HE staining was performed on the ovaries after the intervention. Three rats from each group were randomly selected for ovarian tissue miRNA sequencing, and differential miRNAs were subjected to Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis, and Quantitative Polymerase Chain Reaction (Q-PCR) verification. By using TargetScan to predict the target genes of differential miRNAs, we validated the results with a dual luciferase reporter gene assay.

Results

Compared with the blank group, the estrus cycle of the model group was significantly prolonged (0.01). Anti-Müllerian Hormone (AMH) (0.01) and Estrogen (E2) were significantly decreased (0.05). Follicle-Stimulating Hormone (FSH) (0.05) and Luteinizing Hormone (LH) increased sharply (0.01). Compared with the model group, the estrus cycle was significantly shortened in the acupuncture group (0.01). AMH and E2 were markedly raised (0.05). FSH (0.05) and LH (0.01) were significantly declined. miRNA sequencing showed that there were 23 miRNAs significantly different between the model group and the blank group (0.05), and 30 miRNAs significantly different between the acupuncture group and the model group (0.05). GO demonstrated that the network was mainly involved in cellular components, cells, cellular metabolic processes, binding, and single biological processes; KEGG signaling pathway enrichment showed that it was mainly related to MAPK, adhesion junction, calcium signaling pathways, . Q-PCR results showed that after modeling, the expression rose, and after acupuncture, the expression of the following miRNA decreased:miR-154-5p (0.01), miR-300-5p ( < 0.05), miR-376c-5p (0.05). The dual luciferase reporter assay showed that the relative luciferase activity of the miR-300-5p mimics+MAP3K1-WT group was significantly lower than that of the NC+MAP3K1-WT group (0.01). HE results demonstrated that the number of primordial follicles and primary follicles in the model group was significantly lower than that in the blank group (0.05). Moreover, the morphology was poorer, and the granulosa cell layer was thinner. Compared with the model group, the number of primary follicles in the acupuncture group increased (0.05); the morphology and granulosa cell structure of the ovary were improved to different degrees, and mature follicles could be seen.

Conclusion

Acupuncture may improve the ovarian responsiveness of POR rats by regulating miR-154-5p,miR-300-5p, and miR-376c-5p. Furthermore, miR-300-5p can specifically bind to the 3'-UTR of MAP3K1, and MAP3K1 may be the target of miR-300-5p.

© 2025 Bentham Science Publishers
Loading

Article metrics loading...

/content/journals/cchts/10.2174/0113862073365843241223093834
2025-05-01
2025-09-16

  • From This Site

    /content/journals/cchts/10.2174/0113862073365843241223093834
    dcterms_title,dcterms_subject,pub_keyword
    -contentType:Contributor -contentType:Concept -contentType:Institution
    10
    5
Loading full text...

Full text loading...

References

  1. ZhangJ. JiaH. DiaoF. MaX. LiuJ. CuiY. Efficacy of dehydroepiandrosterone priming in women with poor ovarian response undergoing IVF/ICSI: A meta-analysis.Front. Endocrinol. (Lausanne)202314115628010.3389/fendo.2023.1156280 37361534
     [Google Scholar]
  2. QiQ.R. WuJ. WangY.Q. Clinical effects of adding human recombinant luteinizing hormone at different timing to boreas 4 group patients under antagonist protocol.J. Reprod. Med.20243307852859
     [Google Scholar]
  3. ShaoJ.Y. ShiY. CaiK.X. Treatment of ovarian low response based on the theory of heart-kidney interaction.J. Beijing Uni. Chinese Med.2024470811261130
     [Google Scholar]
  4. GaoS. NingY.C. YaoY.H. Comparative effectiveness of antagonist protocol and PPOS protocol in IVF-ET for women with low ovarian response.J. Clin. Lab. Anal. Med.2024211014151419
     [Google Scholar]
  5. XuJ. ShiJ.Z. ZhangL.T. Oocytes and female age: The influence of ovarian low response on the loss of pregnancy and perinatal risks after single embryo transfer in IVF/ICSI.J. Reprod. Med.20223116461653
     [Google Scholar]
  6. HeX.L. ZhouY.X. LiuL. Clinical efficacy of acupuncture combined with Yulinzhu on patients with ovarian low response undergoing in vitro fertilization-embryo transfer: A randomized controlled trial.Zhongchengyao202446310531056
     [Google Scholar]
  7. FuY.M. YeX.Y. ZouH. A randomized controlled study on the treatment of ovarian low responsiveness infertility with methods of strengthening spleen and kidney.Chin. J. Integr. Med.2024446660664
     [Google Scholar]
  8. XuX.Y. YuG. ChengL.L. Study on the application of different ovarian stimulation protocols in high-age women with low ovarian reserve.Zhongguo Shiyong Fuke Yu Chanke Zazhi202238121219122310.195/38/j.fk2022120115
     [Google Scholar]
  9. ChenY. YinC.X. WangY.Z. Effect of different ovarian stimulation protocols on pregnancy outcomes in patients with low ovarian response undergoing in vitro fertilization-embryo transfer: A comparative study.Chinese J. Sexol.202433106064
     [Google Scholar]
  10. ChenQ.Y. ZhangY. ChenL. Research progress on ovarian stimulation strategies for patients with low response to Poseidon group 4.J. Reprod. Med.2024330912681273
     [Google Scholar]
  11. FerrarettiA.P. La MarcaA. FauserB.C.J.M. TarlatzisB. NargundG. GianaroliL. ESHRE consensus on the definition of ‘poor response’ to ovarian stimulation for in vitro fertilization: The Bologna criteria.Hum. Reprod.20112671616162410.1093/humrep/der092 21505041
     [Google Scholar]
  12. ByersS.L. WilesM.V. DunnS.L. TaftR.A. Mouse estrous cycle identification tool and images.PLoS One201274e3553810.1371/journal.pone.0035538 22514749
     [Google Scholar]
  13. LiuG.M. SunJ.H. LiJ.L. Advances in clinical and animal experiment research on kidney deficiency syndrome.Chinese J. Exper. Formulae2024302326928010.134/22/j.cnki.syfjx.202416
     [Google Scholar]
  14. NieL.N. TangZ.X. HongM.Y. Comparison of the outcomes and clinical pregnancy outcomes of microstimulation and antagonist protocols in patients with low ovarian response.China Pharm.202433159295
     [Google Scholar]
  15. DongL. HuG.H. WangC.Y. Consensus on the clinical diagnosis and treatment of infertility in women with diminished ovarian reserve combined with traditional Chinese medicine and assisted reproductive technology.Shanghai J. Tradit. Chinese Med.202458S1828710.16305/j.1007‑1334.2024.16
     [Google Scholar]
  16. FengY.X. YangH.S. GouM.H. Systematic review on the effectiveness of acupuncture on pregnancy outcomes and ovarian function in women with low ovarian response.World J. Tradit. Chin. Med.2023184491495
     [Google Scholar]
  17. ZhuX.Y. Study on DNA Methylation of IVF-derived Oocytes in Low Responders Improved by Acupuncture.Chengdu University of TCM202110.26988/d.cnki.gcdzu.2021.000023
     [Google Scholar]
  18. LiZ.M. Mechanistic Study on Improving Oocyte Quality of Ovarian Low Response Model Mice by Acupuncture Based on scBS-seq.Chengdu University of TCM202210.26988/d.cnki.gcdzu.2022.000041
     [Google Scholar]
  19. HaoJ.H. WangH.Y. CaoY.X. Effect of Zhi Bian water drainage needling on the PI3K/AKT/FOXO3a pathway in low responsiveness ovarian mice.Zhongguo Zhenjiu2024440782183010.13703/j.0
     [Google Scholar]
  20. WangR.R. SuM-H. LiuL-Y. LaiY-Y. GuoX-L. GanD. ZhengX-Y. YangH. YuS-Y. LiangF-R. WeiW. ZhongY. YangJ. Systematic review of acupuncture to improve ovarian function in women with poor ovarian response.Front. Endocrinol. (Lausanne)202314102885310.3389/fendo.2023.1028853 36992800
     [Google Scholar]
  21. XieZ. PengZ. YaoB. ChenL. MuY. ChengJ. LiQ. LuoX. YangP. XiaY. The effects of acupuncture on pregnancy outcomes of in vitro fertilization: A systematic review and meta-analysis.BMC Complement. Altern. Med.201919113110.1186/s12906‑019‑2523‑7 31200701
     [Google Scholar]
  22. YangL. YangW. SunM. LuoL. LiH.R. MiaoR. PangL. ChenY. ZouK. Meta analysis of ovulation induction effect and pregnancy outcome of acupuncture & moxibustion combined with clomiphene in patients with polycystic ovary syndrome.Front. Endocrinol. (Lausanne)202314126101610.3389/fendo.2023.1261016 38075051
     [Google Scholar]
  23. ChengJ. JinX. ShenJ. MuY. LiQ. XiaL. GaoY. XiaY. Whole transcriptome sequencing reveals how acupuncture and moxibustion increase pregnancy rate in patients undergoing in vitro fertilization-embryo transplantation.BioMed Res. Int.201920191810.1155/2019/4179617 31275970
     [Google Scholar]
  24. DaiY.Y. ZhangX.K. LiH.G. Predictive molecular markers for successful testicular sperm extraction in patients with nonobstructive azoospermia.Chinese J. Androl.2023290985185510.13263/j.cnki.nja.2023.09.015
     [Google Scholar]
  25. WangJ.H. ChangW.H. The function and mechanism of miR-214 in regulating reproductive physiology in animals.Chinese Veterinary J.2024601211111710.20157/j.cnki.zgsyzz.2024.12.016
     [Google Scholar]
  26. HeX.Y. LiuQ.X. ChuX.X. Research progress on the regulation of miRNA on mammalian ovarian follicle development and oocyte maturation.J. Vet. Med. Anim. Health2019501121752185
     [Google Scholar]
  27. IoannidisJ. DonadeuF.X. Circulating miRNA signatures of early pregnancy in cattle.BMC Genomics201617118410.1186/s12864‑016‑2529‑1 26939708
     [Google Scholar]
  28. RongX. ShaoS.C. LiangP. Research progress on the regulation of miRNA in the ovulation process of sheep.J. Livestock Ecol.202344416
     [Google Scholar]
  29. PankiewiczK. LaudańskiP. IssatT. The role of noncoding RNA in the pathophysiology and treatment of premature ovarian insufficiency.Int. J. Mol. Sci.20212217933610.3390/ijms22179336 34502244
     [Google Scholar]
  30. AmbrosV. The functions of animal microRNAs.Nature2004431700635035510.1038/nature02871 15372042
     [Google Scholar]
  31. Salas-HuetosA. JamesE.R. AstonK.I. JenkinsT.G. CarrellD.T. YesteM. The expression of m miRNAs in human ovaries, oocytes, extracellular vesicles, and early embryos: A systematic review.Cells2019812156410.3390/cells8121564 31817143
     [Google Scholar]
  32. CondratC.E. ThompsonD.C. BarbuM.G. BugnarO.L. BobocA. CretoiuD. SuciuN. CretoiuS.M. VoineaS.C. miRNAs as biomarkers in disease: Latest findings regarding their role in diagnosis and prognosis.Cells20209227610.3390/cells9020276 31979244
     [Google Scholar]
  33. ZhangK. ZhongW. LiW.P. ChenZ-J. ZhangC. miR-15a-5p levels correlate with poor ovarian response in human follicular fluid.Reproduction2017154448349610.1530/REP‑17‑0157 28729464
     [Google Scholar]
  34. PAXINOS G. Stereotaxic atlas of rat brain.3rd edBeijingPeople’s Medical Publishing House2005(in Chinese)
     [Google Scholar]
  35. DoughertyJ.P. SpringerD.A. CullenM.J. GershengornM.C. Evaluation of the effects of chemotherapy-induced fatigue and pharmacological interventions in multiple mouse behavioral assays.Behav. Brain Res.201936025526110.1016/j.bbr.2018.12.011 30529403
     [Google Scholar]
  36. HuaS.F. ZhangW.T. An analysis of the mechanism of cangfu daolan decoction in treating polycystic ovary syndrome.Chinese J. Tradit. Clin. Res.202416292834
     [Google Scholar]
  37. YangZ. DengL.H. JiangS.R. Effect of regulating GDF9, c-IAP1/2, JNK1/2, and TNFR1/2 expression by tonifying qi and blood method on granulosa cell apoptosis in superovulated rats. J. Modern Integ.Chinese and Western Med.2024331419051910
     [Google Scholar]
  38. ZhangJ. GaoD.D. GuC.C. Exploring the effect of poria polysaccharides on premature ovarian failure model rats and its molecular mechanism based on MAPK/ERK pathway.Chinese J. Mod. Med.202434134148
     [Google Scholar]
  39. StacheckiJ.J. ArmantD.R. Transient release of calcium from inositol 1,4,5-trisphosphate-specific stores regulates mouse preimplantation development.Development199612282485249610.1242/dev.122.8.2485 8756293
     [Google Scholar]
  40. WangJ. MayernikL. SchultzJ.F. ArmantD.R. Acceleration of trophoblast differentiation by heparin-binding EGF-like growth factor is dependent on the stage-specific activation of calcium influx by ErbB receptors in developing mouse blastocysts.Development20001271334410.1242/dev.127.1.33 10654598
     [Google Scholar]
  41. YangY.Y. YangY.L. LiuW.W. Research progress on the mechanism of regulating inflammation factor-related signaling pathways in the treatment of polycystic ovary syndrome with traditional Chinese medicine [J/OL] Global.Tradit.l Chinese Med.2024112
     [Google Scholar]
  42. ZhaoS.Y. ShenQ. FuG.B. Effect of Zhenfu therapy on blood glucose, insulin levels and p38 mitogen-activated protein kinase protein expression in rats with polycystic ovary syndrome.J. Global Tradit. Chinese Med.2023161019531958
     [Google Scholar]
  43. WangY. ZhangF. Comparative study of three methods for modeling premature ovarian failure in rats.Chin. J. Integr. Med.2022421012251230
     [Google Scholar]
  44. LiZ.M. Study on the Mechanism of Improving Oocyte Quality in Ovarian Low Response Model Mice by Acupuncture Based on scBS-seq.Chengdu University of TCM2022
     [Google Scholar]
  45. LiuM. ZhangD. ZhouX. DuanJ. HuY. ZhangW. LiuQ. XuB. ZhangA. Cell-free fat extract improves ovarian function and fertility in mice with premature ovarian insufficiency.Stem Cell Res. Ther.202213132010.1186/s13287‑022‑03012‑w 35842669
     [Google Scholar]
  46. HaoJ.H. ZhaoY.M. LanY. Protective mechanism of acupuncture on the ovary of low response ovary mice.Res. Acupuncture202449101001100910.13702/j.1000‑0607.20230337 39433360
     [Google Scholar]
  47. WangX.T. The Role of Hsa-circ-0000276 Targeting miR-154-5p/CUL2 Pathway in Cervical Cancer Cells.Shanxi Medical University2023
     [Google Scholar]
  48. ZhaoW. LiuY. ZhangL. DingL. LiY. ZhangH. WangT. HaoM. MicroRNA-154-5p regulates the HPV16 E7-pRb pathway in cervical carcinogenesis by targeting CUL2.J. Cancer202011185379538910.7150/jca.45871 32742484
     [Google Scholar]
  49. ZhangH. WangX. LiY. BaiY. LiQ. WangS. WeiY. LiJ. WenS. ZhaoW. The hsa_circ_0000276-ceRNA regulatory network and immune infiltration in cervical cancer.BMC Cancer202323122210.1186/s12885‑023‑10636‑5 36894874
     [Google Scholar]
  50. LiY. WeiY. ZhangH. BaiY. WangX. LiQ. LiuY. WangS. WangJ. WenS. LiJ. ZhaoW. MicroRNA-154-5p suppresses cervical carcinoma growth and metastasis by silencing Cullin2 in vitro and in vivo.PeerJ202311e1564110.7717/peerj.15641 37397007
     [Google Scholar]
  51. FengL. ChenX. WangY. The correlation between PCNA and MDM2 expression levels in breast cancer tissues and clinicopathological features and HPV16/18 infection.Chinese J. Hosp. Infect. Cont.20233302229232
     [Google Scholar]
  52. ChiJ.R. YuZ.H. LiuB.W. ZhangD. GeJ. YuY. CaoX.C. SNHG5 promotes breast cancer proliferation by sponging the miR-154-5p/PCNA axis.Mol. Ther. Nucleic Acids20191713814910.1016/j.omtn.2019.05.013 31255976
     [Google Scholar]
  53. LiY.H. TianQ. HanQ.C. The effect of microRNA-154-5p on the biological activity of osteosarcoma cells by targeting E2F5 and its mechanism.Basic and Clinical Tumor2024372131134
     [Google Scholar]
  54. LiT.T. ZhuX.Y. LiS.Y. Research progress on the role and mechanism of miRNAs regulating E2F5 in cancer development and progression.Modern Oncol.20233102367373
     [Google Scholar]
  55. GilletV. OuelletA. StepanovY. RodosthenousR.S. CroftE.K. BrennanK. AbdelouahabN. BaccarelliA. TakserL. miRNA profiles in extracellular vesicles from serum early in pregnancies complicated by gestational diabetes mellitus.J. Clin. Endocrinol. Metab.2019104115157516910.1210/jc.2018‑02693 31058973
     [Google Scholar]
  56. LiuX.J. WangJ. ZhangJ. Effect of Linc00152 targeting miR-376c-3p on the proliferation, apoptosis and radiosensitivity of cervical cancer cells.Chinese J. Pathophysiol.202036016774
     [Google Scholar]
  57. JiaoY. ShiY. HanW.C. The clinical significance of lncRNA THRIL and miR-300 in neonatal sepsis patients.Chinese J. Mater. Child Heal. Res.202233084752
     [Google Scholar]
  58. JinX.H. ShiX. LuX. Research progress of microRNAs in acute lung injury.J. Tongji Uni.20214204554561
     [Google Scholar]
  59. YangY.C. YanH.Y. YuanY. Research progress on the regulation of functional components in foods on the activity of the body mediated by microRNA.Chinese J. Food Sci.2022220336136810.16429/j.1009‑7848
     [Google Scholar]
  60. ShiY.N. RenT.C. ZhuW. Study on the effect of miR-300-3p on cell autophagy and H9c2 cardiomyocyte damage induced by hypoxia/reoxygenation.Chin. J. Clin. Pharmacol.2022381618681872
     [Google Scholar]
  61. ZhouS.C. Mechanism Study on the Regulation of Glioblastoma Stem Cell Invasion by MAP3K1/c-JUN Axis.Henan University2022
     [Google Scholar]
  62. ZhouS. NiuR. SunH. KimS.H. JinX. YinJ. The MAP3K1/c-JUN signaling axis regulates glioblastoma stem cell invasion and tumor progression.Biochem. Biophys. Res. Commun.202261218819510.1016/j.bbrc.2022.04.057 35567901
     [Google Scholar]
  63. HuangS.D. ZhangY. ZhangD.N. Exploring the mechanism of traditional chinese medicine treatment for premature ovarian insufficiency based on oxidative stress.Inform. Tradit. Chinese Med.20244106283410.19656/j.cnki.1002‑2406.2024.0605
     [Google Scholar]
  64. WangY.S. The Effect and Mechanism of Puerarin on Polycystic Ovary Syndrome in Mice.Jilin University202410.27162/d.cnki.gjlin.2024.000554
     [Google Scholar]
  65. LiY. ZhangY.S. YouD. Research progress on the role of oxidative stress in ovarian dysfunction [J/OL] Chinese J. Laborat.Animals2024110[Available from: http://kns.cnki.net/kcms/detail/11.2986.Q.2024
    [Google Scholar]
  66. LiX.M. ShiC.X. ChenZ. Expression of TRIP6 protein and mRNA in leiomyoma of uterus.J. Clin. Exp. Pathol.20233908988990
     [Google Scholar]
  67. GengY. QiuY. LiuX. ChenX. DingY. LiuS. ZhaoY. GaoR. WangY. HeJ. Sodium fluoride activates ERK and JNK via induction of oxidative stress to promote apoptosis and impairs ovarian function in rats.J. Hazard. Mater.2014272758210.1016/j.jhazmat.2014.03.011 24681588
     [Google Scholar]
  68. YaoR. TianL.Y. WangF. Research overview of the causes and acupuncture mechanism of premature ovarian insufficiency. Chinese J. Traditional Med.Clinic202436102008201210.164/8/j.cjtcm.2024.1037
     [Google Scholar]
  69. YanJ. ZhaoJ.Y. YinL.W. Effect of “Right Water Passage” acupuncture on the expression of apoptosis-related factors in early-onset ovarian dysfunction rats based on oxidative stress.Zhongguo Zhenjiu20234304454460 37068824
     [Google Scholar]
  70. JiangX. TaiH. XiaoX. ZhangS. CuiS. QiS. HuD. ZhangL. KuangJ. MengX. LiS. Cangfudaotan decoction inhibits mitochondria-dependent apoptosis of granulosa cells in rats with polycystic ovarian syndrome.Front. Endocrinol. (Lausanne)20221396215410.3389/fendo.2022.962154 36465612
     [Google Scholar]
  71. WangC.Y. HuaZ.J. YuJ. The effect of Cangfu Daolan Decoction on the morphology and follicular development of ovaries in a rat model of polycystic ovary syndrome induced by Letrozole.Chinese J. Mod. Appl. Pharm.2022392025932599
     [Google Scholar]
  72. GuoR.L. ZhaoX.W. WangY. Effect of ZiYin ShuLian Tang on apoptosis of granulosa cells in ovarian reserve dysfunction model mice.Anhui Med. J.2024281020872093
     [Google Scholar]
  73. LiY. LiM.F. LiuJ. Study on the protective mechanism of Huyang Yangkun Formula on granulosa cells through the JNK-p38/P65-NF-κB pathway.Zhishi Guoyi Guoyao2022332281285
     [Google Scholar]
  74. ZhuS.X. Pathological Observation of Radon Inhalation-Induced Lung Cancer in BALB/c Mice and Regulatory Mechanism Mediated by Map3k1 Gene.Soochow University2019
     [Google Scholar]
/content/journals/cchts/10.2174/0113862073365843241223093834
Loading
Exploring the Mechanism of Acupuncture in Improving Ovarian Function in Rats with Poor Ovarian Response Using High-Throughput Sequencing
Comb Chem High Throughput Screen 28, 1443 (2025); https://doi.org/10.2174/0113862073365843241223093834
/content/journals/cchts/10.2174/0113862073365843241223093834
/content/journals/cchts/10.2174/0113862073365843241223093834
Loading

Data & Media loading...


  • Article Type:     Research Article
Keyword(s): Acupuncture; granulosa cell structure; MAPK signal path; miRNA; ovarian function; poor ovarian response

Most Cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error
Please enter a valid_number test