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Decreased expression of c-kit and telomerase in a rat model of chronic endometrial ischemia.

Hu J, Yuan R - Med. Sci. Monit. (2011)

Bottom Line: Pou5f1and c-kit mRNA was examined by qPCR.C-kit, caspase3 and telomerase were detected by Western blot.The expression level of caspase3 was significantly higher in the experimental group compared with that in the control group (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, 1st Affiliated Hospital, Chongqing Medical University, Chongqing, China.

ABSTRACT

Background: It was unclear whether chronic endometrial ischemia contributed to the pathogenesis of thin endometrium and was associated with decreased endometrial stem/progenitor cell. Thus, we explored the role of chronic endometrial ischemia in the pathogenesis of thin endometrium and its effect on endometrial stem/progenitor cells apoptosis.

Material/methods: In vitro, endometrial side population (ESP) cell apoptosis models were built, and apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, pou5f1, and c-kit mRNA was detected by qPCR. In vivo, a rat model of chronic endometrial ischemia was induced by performing bilateral uterine artery ligation. TERT and caspase3 were detected by immunohistochemistry. Pou5f1and c-kit mRNA was examined by qPCR. C-kit, caspase3 and telomerase were detected by Western blot.

Results: In the in vitro endometrial SP (ESP) cells apoptosis model, we found that the apoptotic rate was gradually increased with time, prolonging the expression of TERT, and c-kit mRNA was gradually decreased. In the in vivo endometrial SP (ESP) cells apoptosis model, we found that endometrial thickness, luminal epithelium thickness, gland epithelium thickness and the number of glands in the experiment group were significantly decreased compared with those in the control group (P<0.05). The expression levels of c-kit, pou5f1 and telomerase was significantly lower in the experimental group than those in the control group (P<0.05). The expression level of caspase3 was significantly higher in the experimental group compared with that in the control group (P<0.05).

Conclusions: The present work shows that chronic ischemia and chronic endometrial ischemia-associated stem/progenitor cells apoptosis may be responsible for the pathogenesis of thin endometrium.

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Related in: MedlinePlus

Serum deprivation and hypoxia trigger apoptosis in endometrial SP (ESP) cells. Endometrial SP (ESP) cells were incubated for 2, 4, and 6 hours in serum deprivation and hypoxia. Apoptosis was detected by flow cytometry. (A, B): Apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, as confirmed by Annexin V and propidine iodide (PI). (C, D) The relative expression of TERT and c-kit mRNA was detected by qPCR. Data given as mean ±SEM. *p<0.05, vs. control group; Error bars, SEM.
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f1-medscimonit-17-4-br103: Serum deprivation and hypoxia trigger apoptosis in endometrial SP (ESP) cells. Endometrial SP (ESP) cells were incubated for 2, 4, and 6 hours in serum deprivation and hypoxia. Apoptosis was detected by flow cytometry. (A, B): Apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, as confirmed by Annexin V and propidine iodide (PI). (C, D) The relative expression of TERT and c-kit mRNA was detected by qPCR. Data given as mean ±SEM. *p<0.05, vs. control group; Error bars, SEM.

Mentions: During ischemia many changes contribute to cellular death. To explore the effect of these stimuli, endometrial SP (ESP) cells were exposed to culture conditions including serum deprivation and hypoxia. Figures 1A and B show the percentage of endometrial SP (ESP) cells apoptotic cell number in response to serum deprivation and hypoxia treatment (2–6 hours). The flow cytometric detection indicated that along with increased time the apoptotic rate gradually increased. The real-time qPCR assay revealed that along with time prolonging, the expression of TERT and c-kit mRNA was gradually decreased (P<0.05; Figures 1C and D). These data suggest that serum deprivation and hypoxia, both components of ischemia in vivo, induced apoptosis in endometrial SP (ESP) cells. As a result of these findings, we adopted the following experiments, an in vivo model of endometrial SP (ESP) cells apoptosis to confirm the impact of ischemia on the endometrial stem cell population in vivo.


Decreased expression of c-kit and telomerase in a rat model of chronic endometrial ischemia.

Hu J, Yuan R - Med. Sci. Monit. (2011)

Serum deprivation and hypoxia trigger apoptosis in endometrial SP (ESP) cells. Endometrial SP (ESP) cells were incubated for 2, 4, and 6 hours in serum deprivation and hypoxia. Apoptosis was detected by flow cytometry. (A, B): Apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, as confirmed by Annexin V and propidine iodide (PI). (C, D) The relative expression of TERT and c-kit mRNA was detected by qPCR. Data given as mean ±SEM. *p<0.05, vs. control group; Error bars, SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3539516&req=5

f1-medscimonit-17-4-br103: Serum deprivation and hypoxia trigger apoptosis in endometrial SP (ESP) cells. Endometrial SP (ESP) cells were incubated for 2, 4, and 6 hours in serum deprivation and hypoxia. Apoptosis was detected by flow cytometry. (A, B): Apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, as confirmed by Annexin V and propidine iodide (PI). (C, D) The relative expression of TERT and c-kit mRNA was detected by qPCR. Data given as mean ±SEM. *p<0.05, vs. control group; Error bars, SEM.
Mentions: During ischemia many changes contribute to cellular death. To explore the effect of these stimuli, endometrial SP (ESP) cells were exposed to culture conditions including serum deprivation and hypoxia. Figures 1A and B show the percentage of endometrial SP (ESP) cells apoptotic cell number in response to serum deprivation and hypoxia treatment (2–6 hours). The flow cytometric detection indicated that along with increased time the apoptotic rate gradually increased. The real-time qPCR assay revealed that along with time prolonging, the expression of TERT and c-kit mRNA was gradually decreased (P<0.05; Figures 1C and D). These data suggest that serum deprivation and hypoxia, both components of ischemia in vivo, induced apoptosis in endometrial SP (ESP) cells. As a result of these findings, we adopted the following experiments, an in vivo model of endometrial SP (ESP) cells apoptosis to confirm the impact of ischemia on the endometrial stem cell population in vivo.

Bottom Line: Pou5f1and c-kit mRNA was examined by qPCR.C-kit, caspase3 and telomerase were detected by Western blot.The expression level of caspase3 was significantly higher in the experimental group compared with that in the control group (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, 1st Affiliated Hospital, Chongqing Medical University, Chongqing, China.

ABSTRACT

Background: It was unclear whether chronic endometrial ischemia contributed to the pathogenesis of thin endometrium and was associated with decreased endometrial stem/progenitor cell. Thus, we explored the role of chronic endometrial ischemia in the pathogenesis of thin endometrium and its effect on endometrial stem/progenitor cells apoptosis.

Material/methods: In vitro, endometrial side population (ESP) cell apoptosis models were built, and apoptosis was quantified by fluorescence-activated cell sorter (FACS) analysis, pou5f1, and c-kit mRNA was detected by qPCR. In vivo, a rat model of chronic endometrial ischemia was induced by performing bilateral uterine artery ligation. TERT and caspase3 were detected by immunohistochemistry. Pou5f1and c-kit mRNA was examined by qPCR. C-kit, caspase3 and telomerase were detected by Western blot.

Results: In the in vitro endometrial SP (ESP) cells apoptosis model, we found that the apoptotic rate was gradually increased with time, prolonging the expression of TERT, and c-kit mRNA was gradually decreased. In the in vivo endometrial SP (ESP) cells apoptosis model, we found that endometrial thickness, luminal epithelium thickness, gland epithelium thickness and the number of glands in the experiment group were significantly decreased compared with those in the control group (P<0.05). The expression levels of c-kit, pou5f1 and telomerase was significantly lower in the experimental group than those in the control group (P<0.05). The expression level of caspase3 was significantly higher in the experimental group compared with that in the control group (P<0.05).

Conclusions: The present work shows that chronic ischemia and chronic endometrial ischemia-associated stem/progenitor cells apoptosis may be responsible for the pathogenesis of thin endometrium.

Show MeSH
Related in: MedlinePlus