Indole-3-methanol alleviates cisplatin-induced ovarian damage by inhibiting ovarian fibrosis through the TGF-β1/Smad pathway

Abstract

Background Ovarian injury is one of the side effects of chemotherapy in female patients, which seriously endangers female reproductive health. Indole-3-carbinol (I3C), a natural substance abundant in cruciferous vegetables, has been reported to attenuate tissue damage. This study aimed to investigate whether I3C could prevent from ovarian damage induced by chemotherapy. Methods A mouse model of ovarian damage was established by intraperitoneal injection of cisplatin or co-treatment with I3C. Then the ovarian index and estrous cycle was assessed. Meanwhile, follicles counting was conducted to evaluate the effect of I3C in follicular development. Also, we performed the TUNEL and IHC staining to analyze the level of apoptosis and fibrosis, respectively. Western blot and qRT-PCR was used as quantitative methods to evaluate the expression of relative markers and TGF-β1/Smad pathway. Hela cells and Caski cells was used to investigate the anti-tumor activity of I3C by cell counting kit-8, the wound healing assay and colony formation assay in vitro. Results Our results showed that administration of I3C restored the ovary index and improved estrous cycle disorders. Follicle counting results showed that I3C is able to inhibit primordial follicles over-activation caused by cisplatin treatment, and maintained primordial follicle pool. We also found that I3C can down-regulate the levels of Bax and γH2ax, and inhibit the apoptosis of ovarian granulosa cells. In addition, I3C also reduced ovarian fibrosis and inhibited α-SMA and Collagen I expression levels. Further research revealed that I3C treatment significantly down-regulated the activity of the TGF-β1/smad signaling pathway. Finally, we demonstrated that I3C could inhibit the proliferation, migration and colony formation of cervical cancer cells in vitro. Conclusions In summary, I3C alleviates primordial follicular over-activation, granulosa cell apoptosis and ovarian fibrosis induced by cisplatin, and exhibits antitumor activity. Our study provides an innovative therapeutic strategy for preventing ovarian function from chemotherapy in female cancer patients.