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Encircling granulosa cells protects against di-(2-ethylhexyl)phthalate-induced apoptosis in rat oocytes cultured in vitro

Published online by Cambridge University Press:  12 July 2019

Anima Tripathi Open the ORCID record for Anima Tripathi [Opens in a new window] ,
Vivek Pandey ,
A.N. Sahu ,
Alok K. Singh  and
Pawan K. Dubey Open the ORCID record for Pawan K. Dubey [Opens in a new window]
Anima Tripathi*
Affiliation:
Department of Zoology, MMV-Banaras Hindu University, Varanasi 221005, India
Vivek Pandey
Affiliation:
Centre for Genetic Disorders, Institute of Science, Banaras Hindu University, Varanasi 221005
A.N. Sahu
Affiliation:
Department of Pharmaceutical Eng. & Tech. IIT-BHU, Varanasi 221005, India
Alok K. Singh
Affiliation:
Department of Medicine, Institute of Medical Science, Banaras Hindu University, Varanasi-221005, India
Pawan K. Dubey
Affiliation:
Centre for Genetic Disorders, Institute of Science, Banaras Hindu University, Varanasi 221005
*
*Address for correspondence: Anima Tripathi, Department of Zoology, MMV-BHU, Varanasi 221005, India. E-mail: niluskc@gmail.com or Pawan K. Dubey, Centre for Genetic Disorders, Institute of Science, Banaras Hindu University, Varanasi 221005, India. Tel: +91 9451890938. E-mail: pkdubey@bhu.ac.in
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Summary

The present study investigated if the presence of encircling granulosa cells protected against di(2-ethylhexyl)phthalate (DEHP)-induced oxidative stress in rat oocytes cultured in vitro. Denuded oocytes and cumulus–oocyte complexes (COCs) were treated with or without various doses of DEHP (0.0, 25.0, 50.0, 100, 200, 400 and 800 μM) in vitro. Morphological apoptotic changes, levels of oxidative stress and reactive oxygen species (ROS), mitochondrial membrane potential, and expression levels of apoptotic markers (Bcl2, Bax, cytochrome c) were analyzed. Our results showed that DEHP induced morphological apoptotic changes in a dose-dependent manner in denuded oocytes cultured in vitro. The effective dose of DEHP (400 µg) significantly (P>0.05) increased oxidative stress by elevating ROS levels and the mitochondrial membrane potential with higher mRNA expression and protein levels of apoptotic markers (Bax, cytochrome c). Encircling granulosa cells protected oocytes from DEHP-induced morphological changes, increased oxidative stress and ROS levels, as well as increased expression of apoptotic markers. Taken together our data suggested that encircling granulosa cells protected oocytes against DEHP-induced apoptosis and that the presence of granulosa cells could act positively towards the survival of oocytes under in vitro culture conditions and may be helpful during assisted reproductive technique programmes.

Keywords

ApoptosisDEHPGranulosa cellsGene expressionOocyteOxidative stress
Type
Research Article
Information
Zygote , Volume 27 , Issue 4 , August 2019 , pp. 203 - 213
Copyright
© Cambridge University Press 2019 

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Footnotes

†

Both authors contributed equally to this study.

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