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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

Chemotherapy reduced body weight of mice and the number of ovarian follicles. A Bar graph illustrating the body weight of mice in the sham and chemoablated (Cy) groups. B H&E staining of ovaries in sham and treatment groups, 7 and 10 days post-induction. Blue arrow indicated primordial follicle; red arrow indicated primary follicle; black arrow indicated secondary follicle; white arrow indicated mature follicle in B-b. C Bar graph representing the number of follicles at different stages of development. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 500 μm (a, c and d), 200 μm in b. Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
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Fig1: Chemotherapy reduced body weight of mice and the number of ovarian follicles. A Bar graph illustrating the body weight of mice in the sham and chemoablated (Cy) groups. B H&E staining of ovaries in sham and treatment groups, 7 and 10 days post-induction. Blue arrow indicated primordial follicle; red arrow indicated primary follicle; black arrow indicated secondary follicle; white arrow indicated mature follicle in B-b. C Bar graph representing the number of follicles at different stages of development. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 500 μm (a, c and d), 200 μm in b. Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)

Mentions: Ovaries were collected from sham and chemoablated groups at 3, 7, and 10 days after chemotherapy, and at 7 and 28 days after hAEC transplantation. Ovaries were fixed in Bouin's solution (containing 5 % acetic acid, 9 % formaldehyde and 0.9 % picric acid), paraffin-embedded and serially sectioned at a thickness of 5 μm. Hematoxylin-eosin (H&E) staining was used to evaluate the morphological structure of the ovaries, which was evaluated using light microscopy. Follicles were categorized and counted in every fifth section of the ovary, in a method as previously described [19, 20]. Briefly, a primordial follicle was defined as GCs surrounding a single fusiform oocyte. A primary follicle was surrounded by at least three granule cells, resulting in a cubic shape, and a secondary follicle appeared surrounded by at least two layers of GCs with no follicular cavity. Mature follicles (Antral follicles) contain at least two GCs and demonstrated evidence of follicular cavity (Fig. 1B-b).Fig. 1


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)

Chemotherapy reduced body weight of mice and the number of ovarian follicles. A Bar graph illustrating the body weight of mice in the sham and chemoablated (Cy) groups. B H&E staining of ovaries in sham and treatment groups, 7 and 10 days post-induction. Blue arrow indicated primordial follicle; red arrow indicated primary follicle; black arrow indicated secondary follicle; white arrow indicated mature follicle in B-b. C Bar graph representing the number of follicles at different stages of development. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 500 μm (a, c and d), 200 μm in b. Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4549019&req=5

Fig1: Chemotherapy reduced body weight of mice and the number of ovarian follicles. A Bar graph illustrating the body weight of mice in the sham and chemoablated (Cy) groups. B H&E staining of ovaries in sham and treatment groups, 7 and 10 days post-induction. Blue arrow indicated primordial follicle; red arrow indicated primary follicle; black arrow indicated secondary follicle; white arrow indicated mature follicle in B-b. C Bar graph representing the number of follicles at different stages of development. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 500 μm (a, c and d), 200 μm in b. Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
Mentions: Ovaries were collected from sham and chemoablated groups at 3, 7, and 10 days after chemotherapy, and at 7 and 28 days after hAEC transplantation. Ovaries were fixed in Bouin's solution (containing 5 % acetic acid, 9 % formaldehyde and 0.9 % picric acid), paraffin-embedded and serially sectioned at a thickness of 5 μm. Hematoxylin-eosin (H&E) staining was used to evaluate the morphological structure of the ovaries, which was evaluated using light microscopy. Follicles were categorized and counted in every fifth section of the ovary, in a method as previously described [19, 20]. Briefly, a primordial follicle was defined as GCs surrounding a single fusiform oocyte. A primary follicle was surrounded by at least three granule cells, resulting in a cubic shape, and a secondary follicle appeared surrounded by at least two layers of GCs with no follicular cavity. Mature follicles (Antral follicles) contain at least two GCs and demonstrated evidence of follicular cavity (Fig. 1B-b).Fig. 1

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