Erzhi Tiangui formula ameliorates ovarian aging by enhancing antioxidant defense via Nrf2/HO-1 signaling.
EZTG shows promise in improving ovarian function in mice through antioxidant pathways, but further research is needed to assess its relevance in humans.
Where it sits
this study against the rest of the hcg (human chorionic gonadotropin) corpusSummary and findings
This study investigated the effects of Erzhi Tiangui Formula (EZTG) on age-related ovarian aging in mice, focusing on the Nrf2/HO-1 antioxidant pathway. A total of 100 C57BL/6J mice were treated with varying doses of EZTG for 10 days, followed by hormonal injections. The study reported improvements in ovarian morphology and oxidative stress markers.
Abstract
Objective This study aimed to investigate the mechanisms through which Erzhi Tiangui Formula (EZTG) mitigates age-related ovarian aging in mice through the Nrf2/HO-1 antioxidant pathway. Methods A total of 100 C57BL/6J mice were divided into young control (YC, 6-8 weeks, n = 20) and aged groups (8-9 months, n = 80). Aged mice were randomized into four subgroups: control, low-(0.05 g/mL), medium- (0.2 g/mL), and high-dose (0.8 g/mL) EZTG groups (n = 20/group). After 10 days of treatment, ovarian morphology and follicular counts were assessed. On day 11, the remaining mice received an intraperitoneal injection of human menopausal gonadotropin (hMG, 15 IU), followed by human chorionic gonadotropin (hCG, 12 IU) 48 h later. 14 h after hCG administration, the mice were euthanized under deep anesthesia via cervical dislocation, and granulosa cells were collected. The following parameters were evaluated: oxidative stress markers (ROS), Nrf2/HO-1 pathway components (Nrf2, HO-1, Keap1), apoptosis factors (Bax, Bcl-2, cleaved Caspase-3), antioxidant enzymes (SOD, CAT, GPX1), and mitochondrial function (mitochondrial membrane potential, MMP; ATP). In vitro, KGN human granulosa cells were subjected to H₂O₂-induced oxidative stress with or without EZTG-containing serum, and Nrf2 was knocked down by siRNA to validate pathway specificity. Results EZTG treatment significantly improved multiple parameters in a dose-dependent manner. Treated mice exhibited restored estrous cyclicity and enhanced follicular development with reduced atresia. At the cellular level, EZTG effectively decreased ROS accumulation while upregulating antioxidant enzymes, including SOD, CAT, and GPX1. Molecular analyses revealed activation of the Nrf2/HO-1 signaling pathway, as evidenced by increased Nrf2 and HO-1 expression alongside Keap1 downregulation. Additionally, EZTG exerted an anti-apoptotic effect, demonstrated by upregulated Bcl-2 expression and downregulated Bax and cleaved Caspase-3. Mitochondrial function assays further revealed that EZTG significantly improved MMP and ATP production in granulosa cells. Notably, the medium- and high-dose groups consistently outperformed the low-dose group across all measured outcomes, reinforcing a clear dose-response relationship. Conclusion EZTG attenuates ovarian aging by coordinated activation of the Nrf2/HO-1 pathway, enhancing antioxidant defenses, inhibiting apoptosis, and restoring mitochondrial function in a dose-dependent manner. These findings provide mechanistic insights into EZTG's potential as a therapeutic intervention for age-related ovarian decline. Clinical trial number not applicable.
Background
The paper addresses the biological question of ovarian aging and the potential mechanisms to ameliorate its effects. Prior research has indicated that oxidative stress plays a significant role in ovarian aging, and Nrf2/HO-1 signaling is a known pathway for enhancing antioxidant defenses. This study is relevant as it explores a traditional formula's effects on this signaling pathway, potentially offering insights into non-pharmacological interventions.
Methods
Not reported in abstract.
Results
Not reported in abstract.
Interpretation
Not reported in abstract.
Key findings
- Not reported in abstract.
Limitations
- Not reported in abstract.