Fois, Marco (2013) Gender differences in human smooth muscle cells and rat liver. Doctoral Thesis.
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Aim: Besides being different in phenotype, males and females respond also differently to drugs thus it is important to known the differences in order to have a better therapeutic appropriateness. We focused on the liver because it exerts a fundamental role in drug metabolism and on vessels because several significant differences exist between male and female in cardiovascular disease.
Methods: Livers were removed from 18 male and 18 female Sprague–Dawley rats (7 weeks old), washed, weighed, and a part was used for immunohistochemical detection of GCL, and another one was homogenized in cold PBS for biochemical determinations.
Human umbilical arteries were collected from boy and girl cords within 24 hours of birth; smooth muscle cell were isolated from tunica media with collagenase and cultured and used at passage 1. RNA was purified from 100% confluent cells that have the typical morphology of the contractile phenotype. cDNA was synthetized and used as template for gene expression studies by real time PCR.
Results: In livers, methionine, glutathione, taurine, malondialdehyde (a marker of lipid peroxidation), and protein oxidation were similar in both sexes, while L -cysteine and H2S levels were signiﬁcantly higher and lower in female rat livers, respectively. Female liver expressed less GCL than the male liver and this was also confirmed by immunohistochemstry. Interestingly, LAMP-1 expression a marker for lysosomal activity was higher in male liver in comparison with female liver. The datum was confirmed by immunofluorescence staining, while other protein such the autophagic markers and autophagic checkpoint: mTOR did not present significant differences in male and female livers.
Our methodology for purifying VSMC cells resulted extremely affordable and effective, so we evaluated sex-gender differences in gene expression of 9 genes using B2m and rpl30 to normalize the other 7 genes. Importantly, ER-alpha, ER-beta gene expression diverged between sexes being more present in male-VSMC than in female ones, thus suggesting the importance of estrogen receptors in determining sex-gender differences.
Conclusion: The previous results suggest that glytamyl cycle is sex-gender dimorphic and this could produce a different metabolism of important drugs. The divergences in gene expression for ERs in VSMC cultures still need investigations to understand the implications on tunica media physiopathology in a sex-gender perspective.
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