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Long-term progestin contraceptives (LTPOC) induce aberrant angiogenesis, oxidative stress and apoptosis in the guinea pig uterus: A model for abnormal uterine bleeding in humans.

Krikun G, Buhimschi IA, Hickey M, Schatz F, Buchwalder L, Lockwood CJ - J Angiogenes Res (2010)

Bottom Line: The presence of E2 significantly decreased these parameters.These changes were associated with highly elevated of the lipid peroxidation product, 8-isoprostane (8-isoP) content in E2+MPA-treated and by nuclear 8-OH-deoxyguanosine (8oxoG) staining compared to all other groups (p < 0.001).Hence, the GP is an excellent model for the study of LTPOC effects on the uterus and will be extremely useful in determining the mechanistic pathways involved in this process which cannot be conducted on humans.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, 06510, USA. graciela.krikun@yale.edu.

ABSTRACT

Background: Irregular uterine bleeding is the major side effect of, and cause for, discontinuation of long-term progestin-only contraceptives (LTPOCs). The endometria of LTPOC-treated women display abnormally enlarged, fragile blood vessels (BV), decreased endometrial blood flow and oxidative stress. However, obtaining sufficient, good quality tissues have precluded elucidation of the mechanisms underlying these morphological and functional vascular changes.

Methods: The current study assessed the suitability of the guinea pig (GP) as a model for evaluating the uterine effects of LTPOC administration. Thus GPs were treated with a transdermal pellet for 21 days and examined for endometrial histology, angiogenic markers as well as markers of oxidative stress and apoptosis.

Results and discussion: We now demonstrate that GP uteri were enlarged by both estradiol (E2) and medroxyprogesterone acetate (MPA) (p < 0.001). Effects of MPA on uterine weight differed significantly depending on E2 levels (p < 0.001), where MPA opposed the E2 effect in combined treatments. Angiogenesis parameters were similarly impacted upon: MPA alone increased BV density (p = 0.036) and BV average area (p = 0.002). The presence of E2 significantly decreased these parameters. These changes were associated with highly elevated of the lipid peroxidation product, 8-isoprostane (8-isoP) content in E2+MPA-treated and by nuclear 8-OH-deoxyguanosine (8oxoG) staining compared to all other groups (p < 0.001). Abnormalities in the E2+MPA group were consistent with chromatin redistribution, nuclear pyknosis, karyolysis and increased apoptosis as observed by a marked increase in TUNEL labeling.

Conclusions: LTPOC exposure alters endometrial vascular and tissue morphology consistent with oxidative stress and apoptosis in a complex interplay with endogenous estrogens. These findings are remarkably similar to in vivo change observed in the human uterus following LTPOC administration. Hence, the GP is an excellent model for the study of LTPOC effects on the uterus and will be extremely useful in determining the mechanistic pathways involved in this process which cannot be conducted on humans.

No MeSH data available.


Related in: MedlinePlus

Vascular morphology: (Top) vWF staining was conducted on formalin fixed, paraffin embedded tissues for placebo. Bar graphs represents the mean +/- standard error of the mean for placebo control (CTL), E2, MPA or E2+MPA. A) Average blood vessel density, B) blood vessel size and C) blood vessel area heterogeneity. Statistical analysis were conducted by a two way ANOVA followed by Student-Newman-Keuls test for the 4 treatment groups. (Bottom) From left to right, representative IHC of vWF for CTL, E2, MPA and E2+MPA (20×).
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Figure 3: Vascular morphology: (Top) vWF staining was conducted on formalin fixed, paraffin embedded tissues for placebo. Bar graphs represents the mean +/- standard error of the mean for placebo control (CTL), E2, MPA or E2+MPA. A) Average blood vessel density, B) blood vessel size and C) blood vessel area heterogeneity. Statistical analysis were conducted by a two way ANOVA followed by Student-Newman-Keuls test for the 4 treatment groups. (Bottom) From left to right, representative IHC of vWF for CTL, E2, MPA and E2+MPA (20×).

Mentions: Figure 3 displays staining for the endothelial marker, vWF in endometria treated with placebo, E2, MPA or E2+MPA. Based on that staining, the following endpoints were analyzed as follows: Blood vessel density (BVD) = area occupied by blood vessel lumen/area analyzed × 100. Blood vessel size (BVS) = average blood vessel diameter for the field analyzed (microns) and Blood vessel heterogeneity (BVH) = relative SD of blood vessel area for the field analyzed (SD/average area × 100).


Long-term progestin contraceptives (LTPOC) induce aberrant angiogenesis, oxidative stress and apoptosis in the guinea pig uterus: A model for abnormal uterine bleeding in humans.

Krikun G, Buhimschi IA, Hickey M, Schatz F, Buchwalder L, Lockwood CJ - J Angiogenes Res (2010)

Vascular morphology: (Top) vWF staining was conducted on formalin fixed, paraffin embedded tissues for placebo. Bar graphs represents the mean +/- standard error of the mean for placebo control (CTL), E2, MPA or E2+MPA. A) Average blood vessel density, B) blood vessel size and C) blood vessel area heterogeneity. Statistical analysis were conducted by a two way ANOVA followed by Student-Newman-Keuls test for the 4 treatment groups. (Bottom) From left to right, representative IHC of vWF for CTL, E2, MPA and E2+MPA (20×).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2874514&req=5

Figure 3: Vascular morphology: (Top) vWF staining was conducted on formalin fixed, paraffin embedded tissues for placebo. Bar graphs represents the mean +/- standard error of the mean for placebo control (CTL), E2, MPA or E2+MPA. A) Average blood vessel density, B) blood vessel size and C) blood vessel area heterogeneity. Statistical analysis were conducted by a two way ANOVA followed by Student-Newman-Keuls test for the 4 treatment groups. (Bottom) From left to right, representative IHC of vWF for CTL, E2, MPA and E2+MPA (20×).
Mentions: Figure 3 displays staining for the endothelial marker, vWF in endometria treated with placebo, E2, MPA or E2+MPA. Based on that staining, the following endpoints were analyzed as follows: Blood vessel density (BVD) = area occupied by blood vessel lumen/area analyzed × 100. Blood vessel size (BVS) = average blood vessel diameter for the field analyzed (microns) and Blood vessel heterogeneity (BVH) = relative SD of blood vessel area for the field analyzed (SD/average area × 100).

Bottom Line: The presence of E2 significantly decreased these parameters.These changes were associated with highly elevated of the lipid peroxidation product, 8-isoprostane (8-isoP) content in E2+MPA-treated and by nuclear 8-OH-deoxyguanosine (8oxoG) staining compared to all other groups (p < 0.001).Hence, the GP is an excellent model for the study of LTPOC effects on the uterus and will be extremely useful in determining the mechanistic pathways involved in this process which cannot be conducted on humans.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, 06510, USA. graciela.krikun@yale.edu.

ABSTRACT

Background: Irregular uterine bleeding is the major side effect of, and cause for, discontinuation of long-term progestin-only contraceptives (LTPOCs). The endometria of LTPOC-treated women display abnormally enlarged, fragile blood vessels (BV), decreased endometrial blood flow and oxidative stress. However, obtaining sufficient, good quality tissues have precluded elucidation of the mechanisms underlying these morphological and functional vascular changes.

Methods: The current study assessed the suitability of the guinea pig (GP) as a model for evaluating the uterine effects of LTPOC administration. Thus GPs were treated with a transdermal pellet for 21 days and examined for endometrial histology, angiogenic markers as well as markers of oxidative stress and apoptosis.

Results and discussion: We now demonstrate that GP uteri were enlarged by both estradiol (E2) and medroxyprogesterone acetate (MPA) (p < 0.001). Effects of MPA on uterine weight differed significantly depending on E2 levels (p < 0.001), where MPA opposed the E2 effect in combined treatments. Angiogenesis parameters were similarly impacted upon: MPA alone increased BV density (p = 0.036) and BV average area (p = 0.002). The presence of E2 significantly decreased these parameters. These changes were associated with highly elevated of the lipid peroxidation product, 8-isoprostane (8-isoP) content in E2+MPA-treated and by nuclear 8-OH-deoxyguanosine (8oxoG) staining compared to all other groups (p < 0.001). Abnormalities in the E2+MPA group were consistent with chromatin redistribution, nuclear pyknosis, karyolysis and increased apoptosis as observed by a marked increase in TUNEL labeling.

Conclusions: LTPOC exposure alters endometrial vascular and tissue morphology consistent with oxidative stress and apoptosis in a complex interplay with endogenous estrogens. These findings are remarkably similar to in vivo change observed in the human uterus following LTPOC administration. Hence, the GP is an excellent model for the study of LTPOC effects on the uterus and will be extremely useful in determining the mechanistic pathways involved in this process which cannot be conducted on humans.

No MeSH data available.


Related in: MedlinePlus