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Ovarian senescence increases liver fibrosis in humans and zebrafish with steatosis.

Turola E, Petta S, Vanni E, Milosa F, Valenti L, Critelli R, Miele L, Maccio L, Calvaruso V, Fracanzani AL, Bianchini M, Raos N, Bugianesi E, Mercorella S, Di Giovanni M, Craxì A, Fargion S, Grieco A, Cammà C, Cotelli F, Villa E - Dis Model Mech (2015)

Bottom Line: In the entire cohort, at multivariate logistic regression, male gender [odds ratio (OR): 1.408, 95% confidence interval (95% CI): 0.779-2.542, P=0.25] vs women at reproductive age was not associated with F2-F4 fibrosis, whereas a trend was observed for menopause (OR: 1.752, 95% CI: 0.956-3.208, P=0.06).In women, menopause (OR: 2.717, 95% CI: 1.020-7.237, P=0.04) was independently associated with F2-F4 fibrosis.Similarly, in overfed zebrafish, old female fish with failing ovarian function [as demonstrated by extremely low circulating estradiol levels (1.4±0.1 pg/µl) and prevailing presence of atretic follicles in the ovaries] developed massive steatosis and substantial fibrosis (comparable with that occurring in males), whereas young female fish developed less steatosis and were totally protected from the development of fibrosis.

View Article: PubMed Central - PubMed

Affiliation: Gastroenterology Unit, Department of Internal Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy.

No MeSH data available.


Related in: MedlinePlus

Hepatic steatosis in overfed zebrafish. (A) Oil Red O staining of liver sections from young males, old males, young females and old female zebrafish at baseline, and after 1 week of overfeeding. Scale bar: 20 µm. The percentage of the area stained by Oil Red O was quantified by using Image J (Schneider et al., 2012) in three sections from five fish per subgroups (supplementary material Fig. S3B). (B) To compare the development of steatosis between different groups, we measured, by using ImageJ (Schneider et al., 2012), the surface area of 30 randomly assigned hepatocytes in three sections from five different fish per subgroup, which had been stained with H&E (Kuroda et al., 2012). Values are expressed as means±s.e.m. The group of young females was compared with each of the other groups (*P<0.0001; **P<0.01). (C) Quantification of liver triglyceride in overfed zebrafish that had been grouped according to age and gender. Final sample number=6 per subgroup. Values are expressed as means±s.e.m. (*P<0.0001 young females vs old males and old females; °P=0.0049 young females vs young males; **P<0.0001, the group of young females was compared with each of the other groups).
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DMM019950F2: Hepatic steatosis in overfed zebrafish. (A) Oil Red O staining of liver sections from young males, old males, young females and old female zebrafish at baseline, and after 1 week of overfeeding. Scale bar: 20 µm. The percentage of the area stained by Oil Red O was quantified by using Image J (Schneider et al., 2012) in three sections from five fish per subgroups (supplementary material Fig. S3B). (B) To compare the development of steatosis between different groups, we measured, by using ImageJ (Schneider et al., 2012), the surface area of 30 randomly assigned hepatocytes in three sections from five different fish per subgroup, which had been stained with H&E (Kuroda et al., 2012). Values are expressed as means±s.e.m. The group of young females was compared with each of the other groups (*P<0.0001; **P<0.01). (C) Quantification of liver triglyceride in overfed zebrafish that had been grouped according to age and gender. Final sample number=6 per subgroup. Values are expressed as means±s.e.m. (*P<0.0001 young females vs old males and old females; °P=0.0049 young females vs young males; **P<0.0001, the group of young females was compared with each of the other groups).

Mentions: Microscopic evaluation of haematoxylin and eosin staining (H&E) of control and overfed zebrafish liver tissue revealed the presence of hepatic steatosis in overfed zebrafish as early as 1 week after the beginning of the overfeeding protocol (supplementary material Fig. S3A). No changes were observed in the liver of control animals throughout the study. To further evaluate whether lipids accumulated in the liver, hepatic tissue was stained with Oil Red O. In overfed zebrafish, most of the cell cytoplasm was occupied by large fat vacuoles, as shown in Fig. 2A. A significant increase in Oil Red O staining was observed in all overfed groups but young females at week 1 compared with respective baseline fish (male young: P=0.033; male old: P=0.009; female young: P=0.283; female old: P=0.014) (supplementary material Fig. S3B). Periodic acid–Schiff staining (PAS) was used to identify the presence of glycogen. Control livers exhibited normal glycogen storage, whereas overfed livers revealed low levels of glycogen confined in the peripheral region of hepatocytes (supplementary material Fig. S3A).Fig. 2.


Ovarian senescence increases liver fibrosis in humans and zebrafish with steatosis.

Turola E, Petta S, Vanni E, Milosa F, Valenti L, Critelli R, Miele L, Maccio L, Calvaruso V, Fracanzani AL, Bianchini M, Raos N, Bugianesi E, Mercorella S, Di Giovanni M, Craxì A, Fargion S, Grieco A, Cammà C, Cotelli F, Villa E - Dis Model Mech (2015)

Hepatic steatosis in overfed zebrafish. (A) Oil Red O staining of liver sections from young males, old males, young females and old female zebrafish at baseline, and after 1 week of overfeeding. Scale bar: 20 µm. The percentage of the area stained by Oil Red O was quantified by using Image J (Schneider et al., 2012) in three sections from five fish per subgroups (supplementary material Fig. S3B). (B) To compare the development of steatosis between different groups, we measured, by using ImageJ (Schneider et al., 2012), the surface area of 30 randomly assigned hepatocytes in three sections from five different fish per subgroup, which had been stained with H&E (Kuroda et al., 2012). Values are expressed as means±s.e.m. The group of young females was compared with each of the other groups (*P<0.0001; **P<0.01). (C) Quantification of liver triglyceride in overfed zebrafish that had been grouped according to age and gender. Final sample number=6 per subgroup. Values are expressed as means±s.e.m. (*P<0.0001 young females vs old males and old females; °P=0.0049 young females vs young males; **P<0.0001, the group of young females was compared with each of the other groups).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4582103&req=5

DMM019950F2: Hepatic steatosis in overfed zebrafish. (A) Oil Red O staining of liver sections from young males, old males, young females and old female zebrafish at baseline, and after 1 week of overfeeding. Scale bar: 20 µm. The percentage of the area stained by Oil Red O was quantified by using Image J (Schneider et al., 2012) in three sections from five fish per subgroups (supplementary material Fig. S3B). (B) To compare the development of steatosis between different groups, we measured, by using ImageJ (Schneider et al., 2012), the surface area of 30 randomly assigned hepatocytes in three sections from five different fish per subgroup, which had been stained with H&E (Kuroda et al., 2012). Values are expressed as means±s.e.m. The group of young females was compared with each of the other groups (*P<0.0001; **P<0.01). (C) Quantification of liver triglyceride in overfed zebrafish that had been grouped according to age and gender. Final sample number=6 per subgroup. Values are expressed as means±s.e.m. (*P<0.0001 young females vs old males and old females; °P=0.0049 young females vs young males; **P<0.0001, the group of young females was compared with each of the other groups).
Mentions: Microscopic evaluation of haematoxylin and eosin staining (H&E) of control and overfed zebrafish liver tissue revealed the presence of hepatic steatosis in overfed zebrafish as early as 1 week after the beginning of the overfeeding protocol (supplementary material Fig. S3A). No changes were observed in the liver of control animals throughout the study. To further evaluate whether lipids accumulated in the liver, hepatic tissue was stained with Oil Red O. In overfed zebrafish, most of the cell cytoplasm was occupied by large fat vacuoles, as shown in Fig. 2A. A significant increase in Oil Red O staining was observed in all overfed groups but young females at week 1 compared with respective baseline fish (male young: P=0.033; male old: P=0.009; female young: P=0.283; female old: P=0.014) (supplementary material Fig. S3B). Periodic acid–Schiff staining (PAS) was used to identify the presence of glycogen. Control livers exhibited normal glycogen storage, whereas overfed livers revealed low levels of glycogen confined in the peripheral region of hepatocytes (supplementary material Fig. S3A).Fig. 2.

Bottom Line: In the entire cohort, at multivariate logistic regression, male gender [odds ratio (OR): 1.408, 95% confidence interval (95% CI): 0.779-2.542, P=0.25] vs women at reproductive age was not associated with F2-F4 fibrosis, whereas a trend was observed for menopause (OR: 1.752, 95% CI: 0.956-3.208, P=0.06).In women, menopause (OR: 2.717, 95% CI: 1.020-7.237, P=0.04) was independently associated with F2-F4 fibrosis.Similarly, in overfed zebrafish, old female fish with failing ovarian function [as demonstrated by extremely low circulating estradiol levels (1.4±0.1 pg/µl) and prevailing presence of atretic follicles in the ovaries] developed massive steatosis and substantial fibrosis (comparable with that occurring in males), whereas young female fish developed less steatosis and were totally protected from the development of fibrosis.

View Article: PubMed Central - PubMed

Affiliation: Gastroenterology Unit, Department of Internal Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy.

No MeSH data available.


Related in: MedlinePlus