Adipose-derived stem cells promote the repair of chemotherapy-induced premature ovarian failure by inhibiting granulosa cells apoptosis and senescence

Abstract

Background Chemotherapeutic drugs, particularly alkylating cytotoxics such as cyclophosphamide (CTX), play an important role to induce premature ovarian failure (POF). Hormone replacement therapy (HRT) is a widely used treatment to improve hormone secretion. However, the long-term HRT increases the risk of breast cancer and cardiovascular disease are attracting concerns. Therefore, there is an urgent need to develop a safe and effective treatment for POF. Method Adipose-derived stem cells (ADSCs) were isolated and identified from human adipose tissue. For POF modeling, CTX were intraperitoneal injected into CTX-acute group, CTX-chronic group, CTX-acute + ADSCs group and CTX-chronic + ADSCs group rats; For transplantation, ADSCs were transplanted into POF rats through tail-vein. The control group rats were injected with PBS. The effects of POF modeling and transplantation were determined by estrous cycle analysis, histopathological analysis, immunohistochemical staining and apoptosis-related marker. To evaluate the effects of ADSC on granulosa cells in vitro, CTX-induced senescent KGN cells were co-cultured with ADSCs, and senescent-related marker expression was investigated by immunofluorescent staining. Results In vivo studies revealed that ADSCs transplantation reduced the apoptosis of ovarian granulosa cells and secretion of follicle-stimulating hormone. The number of total follicles, primordial follicles, primary follicles, and mature follicles and secretion of anti-Müllerian hormone and estradiol (E2) were also increased by ADSCs. The estrous cycle was also improved by ADSC transplantation. Histopathological analysis showed that CTX-damaged ovarian microenvironment was improved by ADSCs. Furthermore, TUNEL staining indicated that apoptosis of granulosa cells was decreased by ADSCs. In vitro assay also demonstrated that ADSC markedly attenuated CTX-induced senescence and apoptosis of granulosa cell. Mechanistically, both in vivo and in vitro experiments proved that ADSC transplantation suppressed activation of the PI3K/Akt/mTOR axis. Conclusion Our experiment demonstrated that a single injection of high-dose CTX was a less damaging chemotherapeutic strategy than continuous injection of low-dose CTX, and tail-vein injection of ADSCs was a potential approach to promote the restoration of CTX-induced POF.

Cellular senescence in the pathogenesis of ovarian dysfunction

“…Consistent with this, the ovaries harvested from adult mice administered cisplatin showed a senescent phenotype. These findings have been replicated by other groups 39,40 . Moreover, senotherapeutics targeting cellular senescence prevent this cisplatin‐induced ovarian injury and improve fertility, which is accompanied by a reduction in the number of senescent cells in the ovary 35 .…”

“…These findings have been replicated by other groups. 39,40 Moreover, senotherapeutics targeting cellular senescence prevent this cisplatininduced ovarian injury and improve fertility, which is accompanied by a reduction in the number of senescent cells in the ovary. 35 The senotherapeutics used in this study were a combination of dasatinib and quercetin (DQ), metformin, and DQ plus metformin, and the results showed that DQ plus metformin was the most effective.…”

Metformin protects ovarian granulosa cells in chemotherapy-induced premature ovarian failure mice through AMPK/PPAR-γ/SIRT1 pathway

“…Generally speaking, after natural drying and using Swiss-Giemsa staining, according to the type and proportion of cells in the vaginal fluid (especially leukocytes, nucleated epithelial cells, and cornified squamous epithelial cells) that had been just collected by an optical microscope, we may infer which estrous phase the mice was in. Normally, the estrous cycle consists of 4 phases, including proestrus, estrus, metestrus, and diestrus 36 . Proestrus smears were dominated by nucleated epithelial cells, while estrus smears were predominantly composed of nucleated keratinocytes.…”

Current Status and Future Prospects of Stem Cell Therapy for Infertile Patients with Premature Ovarian Insufficiency

“…Various studies have explored the mechanisms of action of hADSCs in POI, with paracrine regulation being frequently postulated [72,119,120]. These investigations suggest that downregulation of the phosphoinositide 3-kinases (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) axis and SMAD pathways contributes to the repair of chemotherapy-induced POI in ovaries.…”