Elena Shepel1, Nataliya Grushka2, Nataliya Makogon3, Valentyna Sribna4, Svitlana Pavlovych5, Roman Yanchii6. 1. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: elena-shepel@ukr.net. 2. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: grunay@i.ua. 3. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: nataliya.v.makogon@gmail.com. 4. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: valia-z@ukr.net. 5. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: spavl@biph.kiev.ua. 6. Department of Immunophysiology, Bogomoletz Institute of Physiology, NAS of Ukraine, Kyiv, Ukraine. Electronic address: tas@biph.kiev.ua.
Abstract
BACKGROUND: Lipopolysaccharide (LPS), the endotoxin of gram-negative bacteria, can impair female reproductive function. However, there is a little information about genotoxic stress in ovarian follicular cells as well as about the changes in oocyte developmental potential under endotoxemia. So the aim of our study was to investigate in vitro oocyte maturation, the DNA damage and expression of some developmental competence-related genes in follicular cells of mice treated with LPS. METHODS: LPS (3mg/kg) was intraperitoneally injected into the mice for 24h, and in vitro maturation of mouse oocyte was determined. The expression levels of genes in cumulus cells were detected by reverse transcriptase polymerase chain reaction. DNA damage in granulosa cells was assessed by the alkaline comet assay. RESULTS: LPS injection caused an impairment of oocyte maturation in vitro: the percentage of oocytes reaching metaphase I and metaphase II decreased markedly compared to vehicle control mice. At the same time we observed strong DNA damage in granulosa cells of LPS-treated animals. The endotoxemia resulted in significantly reduced mRNA expression levels for hyaluronan synthase 2 (HAS2), cyclooxygenase 2 (COX2) and Gremlin-1 (GREM1) genes compared with control. CONCLUSIONS: Our results obtained in a mouse model of endotoxin-induced female reproductive dysfunction suggest that LPS may affect oocyte quality through the induction of DNA damage and decreasing the cumulus expression of genes associated with cumulus expansion and oocyte maturation, such as HAS2, COX2 and GREM1.
BACKGROUND:Lipopolysaccharide (LPS), the endotoxin of gram-negative bacteria, can impair female reproductive function. However, there is a little information about genotoxic stress in ovarian follicular cells as well as about the changes in oocyte developmental potential under endotoxemia. So the aim of our study was to investigate in vitro oocyte maturation, the DNA damage and expression of some developmental competence-related genes in follicular cells of mice treated with LPS. METHODS:LPS (3mg/kg) was intraperitoneally injected into the mice for 24h, and in vitro maturation of mouse oocyte was determined. The expression levels of genes in cumulus cells were detected by reverse transcriptase polymerase chain reaction. DNA damage in granulosa cells was assessed by the alkaline comet assay. RESULTS:LPS injection caused an impairment of oocyte maturation in vitro: the percentage of oocytes reaching metaphase I and metaphase II decreased markedly compared to vehicle control mice. At the same time we observed strong DNA damage in granulosa cells of LPS-treated animals. The endotoxemia resulted in significantly reduced mRNA expression levels for hyaluronan synthase 2 (HAS2), cyclooxygenase 2 (COX2) and Gremlin-1 (GREM1) genes compared with control. CONCLUSIONS: Our results obtained in a mouse model of endotoxin-induced female reproductive dysfunction suggest that LPS may affect oocyte quality through the induction of DNA damage and decreasing the cumulus expression of genes associated with cumulus expansion and oocyte maturation, such as HAS2, COX2 and GREM1.
Authors: Błażej Chermuła; Wiesława Kranc; Piotr Celichowski; Bogusława Stelmach; Hanna Piotrowska-Kempisty; Paul Mozdziak; Leszek Pawelczyk; Robert Zygmunt Spaczyński; Bartosz Kempisty Journal: J Clin Med Date: 2021-12-24 Impact factor: 4.241