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siRNA as a tool to delineate pathway channelization in H2O2 induced apoptosis of primary Leydig cells in vitro

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Abstract

Using siRNA as a tool, the channelization of pathway in H2O2 induced apoptosis of primary Leydig cells was investigated in vitro. Exposure (4 h) to H2O2 (250 μM) induced maximum apoptosis but affected Leydig cell viability significantly. Therefore, expression of apoptotic marker genes, caspase-8, -9, -3 and polyadenosine ribose polymerase was subsequently investigated using the same concentration post 1 h exposure. Incubation with siRNA (20 nM) either for caspase-8 or -9, inhibited their individual expressions by 55–60 % and activity, 50–55 %. The inhibition efficiency using siRNA was comparable with post- or pre-H2O2 treatment of cells. Like siRNA, Eugenia jambolana (100 μg/ml) plant extract too, effectively countered over-expression of all apoptotic marker proteins. Silencing expressions of caspase 8 but not 9 through siRNA leads to a profound inhibition of caspase 3 implying that H2O2 induced Leydig cell apoptosis is preferably channeled through extrinsic and later extending to other pathways.

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Acknowledgments

The generous gift of EJE extract from Dr. Suman Bala Sharma, Professor, University College of Medical Sciences, Delhi, is greatly acknowledged. The study was funded by NIHFW.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Baba Gang Nath Marg, Munirka, New Delhi, 110067, India

    Himani Anand & M. M. Misro

  2. Department of Biochemistry, University College of Medical Sciences, University of Delhi, Dilshad Garden, New Delhi, 110095, India

    S. B. Sharma

  3. Department of Zoology, Dayalbagh Educational Institute, Dayalbagh, Agra, 282005, India

    Himani Anand & Sant Prakash

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  1. Himani Anand
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  2. M. M. Misro
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Corresponding author

Correspondence to M. M. Misro.

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Anand, H., Misro, M.M., Sharma, S.B. et al. siRNA as a tool to delineate pathway channelization in H2O2 induced apoptosis of primary Leydig cells in vitro. Apoptosis 17, 1131–1143 (2012). https://doi.org/10.1007/s10495-012-0749-7

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