A traditional Chinese medicine formulation known as Dingkun Pill (DKP) reduced ovarian damage in a rat model of chemotherapy-induced premature ovarian failure, according to a research article published in the journal Biological Research on 27 October 2026.
Its effects on premature ovarian failure and the underlying mechanisms had remained unclear, which motivated the new experiments.
To test the formula, the researchers established a cyclophosphamide-induced premature ovarian failure model in Sprague-Dawley rats. The affected animals were divided into several groups: an untreated POF group, three DKP dose groups receiving 0.45, 0.9, or 1.8 grams per kilogram, a positive control group treated with dehydroepiandrosterone (DHEA), a group receiving the PI3K inhibitor LY294002 alone, and a group receiving the medium DKP dose together with LY294002.
The study protocol was detailed and multi-layered. Changes in the estrous cycle were monitored through vaginal smear examination for 14 consecutive days, beginning on the 14th day of drug intervention. After 28 days of treatment, the rats were euthanized, and serum sex hormone and oxidative stress marker levels were measured with ELISA kits. Ovarian tissue was examined with hematoxylin-eosin staining, while Western blot analysis detected proteins related to autophagy, apoptosis, proliferation, and the PI3K/AKT/mTOR axis. Immunofluorescence was used to assess autophagy, proliferation, and apoptosis status in ovarian granulosa cells.
The results showed a dose-dependent pattern. DKP improved the trend of weight loss in the cyclophosphamide-treated rats, reversed sex hormone disorders, and restored normal estrous cycles. The ovarian protective effect of the highest dose, 1.8 grams per kilogram, was reported to be comparable to that of DHEA, the positive control treatment.
At the medium dose of 0.9 grams per kilogram, DKP alleviated ovarian damage, reduced follicular depletion, and promoted follicular development. The treatment also produced marked changes in oxidative stress markers: serum malondialdehyde fell from 7.7 to 4.3 micromoles per liter, reactive oxygen species dropped from 215.3 to 142.3 units per milliliter, and superoxide dismutase activity rose from 2.3 to 3.2 units per milliliter.
Beyond the oxidative stress findings, the treatment influenced cell-level processes in the ovary. DKP inhibited excessive autophagy in the ovaries of the POF rats, decreased levels of pro-apoptosis-related proteins, increased the proliferative activity of granulosa cells, and up-regulated function-related proteins of these cells, according to the reported results.
The mechanistic findings centered on the PI3K/AKT/mTOR pathway. The observed effects were associated with activation of this signaling axis, and when the PI3K inhibitor LY294002 was co-administered with the medium DKP dose, it partially attenuated the formula’s ovarian protection. This suggests, within the limits of the model, that the pathway contributes to the observed benefits.
The authors concluded that DKP alleviates cyclophosphamide-induced premature ovarian failure in rats by reducing oxidative stress and regulating autophagy and apoptosis, with these effects associated with the PI3K/AKT/mTOR axis. It is important to note that these findings come from a rat model of chemically induced ovarian failure, and the article is an early-release, peer-reviewed accepted version that remains subject to further edits before the final version of record. The results therefore describe animal-model outcomes and pathway associations rather than demonstrated clinical benefits in patients, and further research would be needed to establish any effects in humans.
Subject of Research: Effects of Dingkun Pill on cyclophosphamide-induced premature ovarian failure in rats via the PI3K/AKT/mTOR pathway
Article Title: Dingkun Pill attenuates oxidative stress, autophagy, and apoptosis to alleviate cyclophosphamide-induced premature ovarian failure in rats via activation of the PI3K/AKT/mTOR signaling pathway
Article References: Guan, Y., Guo, J., Sui, X., Zhao, Q., Zhang, Z., & Cui, Z. (2026). Dingkun Pill attenuates oxidative stress, autophagy, and apoptosis to alleviate cyclophosphamide-induced premature ovarian failure in rats via activation of the PI3K/AKT/mTOR signaling pathway. Biological Research. https://doi.org/10.1186/s40659-026-00738-2
Image Credits: AI Generated
DOI: 10.1186/s40659-026-00738-2
Keywords: Dingkun Pill, premature ovarian failure, cyclophosphamide, PI3K/AKT/mTOR, oxidative stress, autophagy, apoptosis, granulosa cells, rats, traditional Chinese medicine, LY294002, DHEA
News Source: Drew Townsend. (October 27, 2026). Traditional Chinese Medicine Formula Shows Ovarian Protection in Rat Study. Scienmag.
