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Human amniotic epithelial cells inhibit granulosa cell apoptosis induced by chemotherapy and restore the fertility.

Zhang Q, Xu M, Yao X, Li T, Wang Q, Lai D - Stem Cell Res Ther (2015)

Bottom Line: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients.Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation. hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation.These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

View Article: PubMed Central - PubMed

Affiliation: International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, 145, Guang-Yuan Road, Shanghai, 200030, People's Republic of China. 10111010022@fudan.edu.cn.

ABSTRACT

Introduction: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients. Recently, stem cell transplantation has been identified as a promising treatment for premature ovarian failure and insufficiency. We have previously demonstrated that human amniotic epithelial cells (hAECs) migrate into injured tissue and promote the recovery of ovarian function in chemoablated mice. However, the molecular mechanism guiding this process remains unclear.

Methods: To further investigate the effect of hAECs on chemotherapy-induced apoptosis, cultured primary hAECs were injected intravenously into mice treated with cyclophosphamide and busulphan. Apoptosis of granulosa cells was observed by TUNEL staining, and apoptosis-related gene expression was performed on ovarian tissue by real-time PCR and Western blot 7 days after hAEC transplantation. Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation.

Results: hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation. In addition, 4 weeks after transplantation, hAECs promoted the development of follicles and increased the number of cumulus oocyte complexes in chemoablated mice. Furthermore, hAECs improved ovarian mass and increased the number of follicles compared to those of the chemoablated group, and hAEC transplantation partially rescued the fertility of chemoablated mice.

Conclusions: hAEC transplantation promotes ovarian function by inhibiting tumor necrosis factor-alpha-mediated cell apoptosis and reducing inflammation in chemotherapy-induced premature ovarian failure. These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

No MeSH data available.


Related in: MedlinePlus

hAEC transplantation reduced chemotherapy-induced inflammation in ovarian tissue. A Section of dissected hAECs illustrated a cobble stone-like epithelial cell morphology. B Relative expression of mRNA levels detected by real-time PCR in cultured hAECs. C Image of fixed ovarian tissue in different groups at 7 days after hAEC transplantation. D Bar graph demonstrated the ovarian weights in different groups. E H&E staining results displaying ovarian morphology in different groups at 7 days after hAEC transplantation. F Relative expression of inflammatory cytokine mRNA levels in injured ovarian tissue analyzed by real-time PCR at 7 days after hAEC transplantation. Data represent means ± SEM; *p < 0.05 versus Sham, #p < 0.05 versus Cy. Scale bar = 200 μm in (E). Cy chemoablated group (n = 6), Cy + hAECs hAEC-treated group (n = 6), Sham sham group (n = 4)
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Fig3: hAEC transplantation reduced chemotherapy-induced inflammation in ovarian tissue. A Section of dissected hAECs illustrated a cobble stone-like epithelial cell morphology. B Relative expression of mRNA levels detected by real-time PCR in cultured hAECs. C Image of fixed ovarian tissue in different groups at 7 days after hAEC transplantation. D Bar graph demonstrated the ovarian weights in different groups. E H&E staining results displaying ovarian morphology in different groups at 7 days after hAEC transplantation. F Relative expression of inflammatory cytokine mRNA levels in injured ovarian tissue analyzed by real-time PCR at 7 days after hAEC transplantation. Data represent means ± SEM; *p < 0.05 versus Sham, #p < 0.05 versus Cy. Scale bar = 200 μm in (E). Cy chemoablated group (n = 6), Cy + hAECs hAEC-treated group (n = 6), Sham sham group (n = 4)

Mentions: To eluciate the impact of hAECs on chemotherapy-induced inflammation, we separated hAECs from the fresh amnion of placenta tissue in vitro. Under light microscopy, the primary hAECs appeared as cobble stone-like epithelial cells (Fig. 3A). Real-time PCR demonstrated that cultured hAECs expressed high mRNA levels of Cytokeratin 19 (CK19), Vimentin, E-cadherin (E-cad) and Octamer-binding transcription factor (Oct-4), indicating that hAECs have the character of epithelia cells and stem cells (Fig. 3B). Although hAEC transplantation had no significant effect on the body weight of mice, ovarian weight in the hAEC-treated group was higher than those of the chemoablated group at 7 days (Fig. 3C and D). Moreover, hAEC-treated mice displayed more follicles than chemoablated mice at 7 days after hAEC transplantation (Fig. 3E). Interleukin (IL)-1ß, a pro-inflammatory cytokine, was significantly increased (by real-time PCR analysis) following chemotherapy, while the anti-inflammatory cytokine (IL-10) was significantly reduced in injured ovarian tissue. However, the increase in IL-1ß expression was inhibited in the hAEC-treated group. These results indicate that grafted hAECs may partially inhibit the chemotherapy-induced inflammatory response to achieve the goal of reduced of ovarian injury.Fig. 3


Human amniotic epithelial cells inhibit granulosa cell apoptosis induced by chemotherapy and restore the fertility.

Zhang Q, Xu M, Yao X, Li T, Wang Q, Lai D - Stem Cell Res Ther (2015)

hAEC transplantation reduced chemotherapy-induced inflammation in ovarian tissue. A Section of dissected hAECs illustrated a cobble stone-like epithelial cell morphology. B Relative expression of mRNA levels detected by real-time PCR in cultured hAECs. C Image of fixed ovarian tissue in different groups at 7 days after hAEC transplantation. D Bar graph demonstrated the ovarian weights in different groups. E H&E staining results displaying ovarian morphology in different groups at 7 days after hAEC transplantation. F Relative expression of inflammatory cytokine mRNA levels in injured ovarian tissue analyzed by real-time PCR at 7 days after hAEC transplantation. Data represent means ± SEM; *p < 0.05 versus Sham, #p < 0.05 versus Cy. Scale bar = 200 μm in (E). Cy chemoablated group (n = 6), Cy + hAECs hAEC-treated group (n = 6), Sham sham group (n = 4)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4549019&req=5

Fig3: hAEC transplantation reduced chemotherapy-induced inflammation in ovarian tissue. A Section of dissected hAECs illustrated a cobble stone-like epithelial cell morphology. B Relative expression of mRNA levels detected by real-time PCR in cultured hAECs. C Image of fixed ovarian tissue in different groups at 7 days after hAEC transplantation. D Bar graph demonstrated the ovarian weights in different groups. E H&E staining results displaying ovarian morphology in different groups at 7 days after hAEC transplantation. F Relative expression of inflammatory cytokine mRNA levels in injured ovarian tissue analyzed by real-time PCR at 7 days after hAEC transplantation. Data represent means ± SEM; *p < 0.05 versus Sham, #p < 0.05 versus Cy. Scale bar = 200 μm in (E). Cy chemoablated group (n = 6), Cy + hAECs hAEC-treated group (n = 6), Sham sham group (n = 4)
Mentions: To eluciate the impact of hAECs on chemotherapy-induced inflammation, we separated hAECs from the fresh amnion of placenta tissue in vitro. Under light microscopy, the primary hAECs appeared as cobble stone-like epithelial cells (Fig. 3A). Real-time PCR demonstrated that cultured hAECs expressed high mRNA levels of Cytokeratin 19 (CK19), Vimentin, E-cadherin (E-cad) and Octamer-binding transcription factor (Oct-4), indicating that hAECs have the character of epithelia cells and stem cells (Fig. 3B). Although hAEC transplantation had no significant effect on the body weight of mice, ovarian weight in the hAEC-treated group was higher than those of the chemoablated group at 7 days (Fig. 3C and D). Moreover, hAEC-treated mice displayed more follicles than chemoablated mice at 7 days after hAEC transplantation (Fig. 3E). Interleukin (IL)-1ß, a pro-inflammatory cytokine, was significantly increased (by real-time PCR analysis) following chemotherapy, while the anti-inflammatory cytokine (IL-10) was significantly reduced in injured ovarian tissue. However, the increase in IL-1ß expression was inhibited in the hAEC-treated group. These results indicate that grafted hAECs may partially inhibit the chemotherapy-induced inflammatory response to achieve the goal of reduced of ovarian injury.Fig. 3

Bottom Line: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients.Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation. hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation.These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

View Article: PubMed Central - PubMed

Affiliation: International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, 145, Guang-Yuan Road, Shanghai, 200030, People's Republic of China. 10111010022@fudan.edu.cn.

ABSTRACT

Introduction: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients. Recently, stem cell transplantation has been identified as a promising treatment for premature ovarian failure and insufficiency. We have previously demonstrated that human amniotic epithelial cells (hAECs) migrate into injured tissue and promote the recovery of ovarian function in chemoablated mice. However, the molecular mechanism guiding this process remains unclear.

Methods: To further investigate the effect of hAECs on chemotherapy-induced apoptosis, cultured primary hAECs were injected intravenously into mice treated with cyclophosphamide and busulphan. Apoptosis of granulosa cells was observed by TUNEL staining, and apoptosis-related gene expression was performed on ovarian tissue by real-time PCR and Western blot 7 days after hAEC transplantation. Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation.

Results: hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation. In addition, 4 weeks after transplantation, hAECs promoted the development of follicles and increased the number of cumulus oocyte complexes in chemoablated mice. Furthermore, hAECs improved ovarian mass and increased the number of follicles compared to those of the chemoablated group, and hAEC transplantation partially rescued the fertility of chemoablated mice.

Conclusions: hAEC transplantation promotes ovarian function by inhibiting tumor necrosis factor-alpha-mediated cell apoptosis and reducing inflammation in chemotherapy-induced premature ovarian failure. These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

No MeSH data available.


Related in: MedlinePlus