The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency
Abstract Background Premature ovarian insufficiency (POI) is a refractory gynecological endocrine disorder. Ningxin-Tongyu-Zishen formula (NTZF), developed based on the ‘simultaneous heart-kidney regulation’ principle, exhibits therapeutic efficacy in treating POI, potentially through regulating pro...
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| Format: | Article |
| Language: | English |
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BMC
2025-08-01
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| Series: | Journal of Ovarian Research |
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| Online Access: | https://doi.org/10.1186/s13048-025-01769-1 |
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| author | Jiawen Ma Chaofan Zhu Lifang Xie Shuaiqi An Zaiyang Zhang Keying Wang Yizhou Zhang |
| author_facet | Jiawen Ma Chaofan Zhu Lifang Xie Shuaiqi An Zaiyang Zhang Keying Wang Yizhou Zhang |
| author_sort | Jiawen Ma |
| collection | DOAJ |
| description | Abstract Background Premature ovarian insufficiency (POI) is a refractory gynecological endocrine disorder. Ningxin-Tongyu-Zishen formula (NTZF), developed based on the ‘simultaneous heart-kidney regulation’ principle, exhibits therapeutic efficacy in treating POI, potentially through regulating proBDNF/mBDNF balance. This study aimed to elucidate the molecular mechanism by which NTZF treats POI via proBDNF/mBDNF modulation. Methods POI rat models were established using cyclophosphamide (CTX). The therapeutic effects of NTZF were evaluated by analyzing estrous cycles, ovarian indices, follicular development, serum sex hormone levels (FSH, E2, AMH), and ovarian granulosa cells (OGCs) apoptosis. Following immunofluorescence staining to localize BDNF receptors, proBDNF/mBDNF protein expression was quantified in brain and ovarian tissues. The active metabolite of CTX, phosphoramide mustard (PM), was employed to induce damage in KGN cells. The regulatory effect of NTZF on proBDNF/mBDNF was investigated and compared with recombinant mBDNF protein. tPA and PAI-1 was screened, and their interactions with NTZF were analyzed. mRNA and protein expression of tPA, PAI-1, and tPA-PAI-1 complexes were assessed via q-PCR and Western Blot. Results NTZF composition was characterized and shown to improve ovarian function in POI rats. Its mechanism involves correcting proBDNF/mBDNF imbalance in both brain and ovarian tissues. NTZF achieved this correction through the PAI-1/tPA signaling pathway, thereby inhibiting apoptosis in damaged KGN cells. Conclusion Our findings demonstrate that NTZF attenuates PAI-1 expression, diminishes tPA-PAI-1 complex formation, and potentiates tPA-dependent proteolysis of proBDNF into mBDNF, thereby restoring their balance. This restores proBDNF/mBDNF balance, suppresses OGCs apoptosis, and ultimately ameliorates POI. |
| format | Article |
| id | doaj-art-0052d96bcdb54f178a347b4235bd12f1 |
| institution | DOAJ |
| issn | 1757-2215 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Ovarian Research |
| spelling | doaj-art-0052d96bcdb54f178a347b4235bd12f12025-08-20T03:05:55ZengBMCJournal of Ovarian Research1757-22152025-08-0118111410.1186/s13048-025-01769-1The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiencyJiawen Ma0Chaofan Zhu1Lifang Xie2Shuaiqi An3Zaiyang Zhang4Keying Wang5Yizhou Zhang6School of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversitySchool of Basic Medical Sciences, Zhejiang Chinese Medical UniversityAbstract Background Premature ovarian insufficiency (POI) is a refractory gynecological endocrine disorder. Ningxin-Tongyu-Zishen formula (NTZF), developed based on the ‘simultaneous heart-kidney regulation’ principle, exhibits therapeutic efficacy in treating POI, potentially through regulating proBDNF/mBDNF balance. This study aimed to elucidate the molecular mechanism by which NTZF treats POI via proBDNF/mBDNF modulation. Methods POI rat models were established using cyclophosphamide (CTX). The therapeutic effects of NTZF were evaluated by analyzing estrous cycles, ovarian indices, follicular development, serum sex hormone levels (FSH, E2, AMH), and ovarian granulosa cells (OGCs) apoptosis. Following immunofluorescence staining to localize BDNF receptors, proBDNF/mBDNF protein expression was quantified in brain and ovarian tissues. The active metabolite of CTX, phosphoramide mustard (PM), was employed to induce damage in KGN cells. The regulatory effect of NTZF on proBDNF/mBDNF was investigated and compared with recombinant mBDNF protein. tPA and PAI-1 was screened, and their interactions with NTZF were analyzed. mRNA and protein expression of tPA, PAI-1, and tPA-PAI-1 complexes were assessed via q-PCR and Western Blot. Results NTZF composition was characterized and shown to improve ovarian function in POI rats. Its mechanism involves correcting proBDNF/mBDNF imbalance in both brain and ovarian tissues. NTZF achieved this correction through the PAI-1/tPA signaling pathway, thereby inhibiting apoptosis in damaged KGN cells. Conclusion Our findings demonstrate that NTZF attenuates PAI-1 expression, diminishes tPA-PAI-1 complex formation, and potentiates tPA-dependent proteolysis of proBDNF into mBDNF, thereby restoring their balance. This restores proBDNF/mBDNF balance, suppresses OGCs apoptosis, and ultimately ameliorates POI.https://doi.org/10.1186/s13048-025-01769-1Premature ovarian insufficiencyNingxin-Tongyu-Zishen formulaProBDNFmBDNFPAI-1tPA |
| spellingShingle | Jiawen Ma Chaofan Zhu Lifang Xie Shuaiqi An Zaiyang Zhang Keying Wang Yizhou Zhang The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency Journal of Ovarian Research Premature ovarian insufficiency Ningxin-Tongyu-Zishen formula ProBDNF mBDNF PAI-1 tPA |
| title | The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency |
| title_full | The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency |
| title_fullStr | The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency |
| title_full_unstemmed | The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency |
| title_short | The mechanism of Ningxin-Tongyu-Zishen formula regulating probdnf/mbdnf balance through PAI-1/tPA signaling pathway in the treatment of premature ovarian insufficiency |
| title_sort | mechanism of ningxin tongyu zishen formula regulating probdnf mbdnf balance through pai 1 tpa signaling pathway in the treatment of premature ovarian insufficiency |
| topic | Premature ovarian insufficiency Ningxin-Tongyu-Zishen formula ProBDNF mBDNF PAI-1 tPA |
| url | https://doi.org/10.1186/s13048-025-01769-1 |
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