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Estrogen Receptor β Mediated Neuroprotective Efficacy of Cicer microphyllum Seed Extract in Global Hypoxia

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Abstract

Hypoxia induced oxidative stress and neurodegeneration in the hippocampus has been implicated for memory impairment in conditions like stroke, ischemia and hypobaric hypoxia. The present study, aimed at investigating the potential of ethanolic extract of Cicer microphyllum seeds (CSE) for amelioration of global hypoxia induced neurodegeneration in CA1 region of hippocampus. CSE supplementation considerably reduced neurodegeneration and dendritic atrophy in CA1 neurons along with improvement of memory in hypoxic rats. This effect of CSE was partly attributed to its antioxidant activity resulting in reduction of lipid peroxidation, protein oxidation and DNA damage during exposure to chronic hypoxia. CSE also promoted dendritic arborization through activation of estrogen receptor beta (ERβ) and phosphorylation of extracellular signal regulated kinase (ERK1/2) which was independent of brain derived neurotrophic factor (BDNF) mediated signalling mechanisms. Extra nuclear activation of ERK1/2 by ERβ resulted in phosphorylation of cyclic AMP response element binding protein (CREB) leading to increased expression of PSD-95.These molecular alterations translated to behavioural changes in CSE administered hypoxic animals that performed better in Morris Water Maze Task as compared to vehicle treated hypoxic animals. Toxicological studies show NOEAL > 2000 mg/kg b.w. for oral administration of CSE indicating its safety for consumption. Our findings not only suggest the neuroprotective potential of CSE in hypoxia but also provide evidence for involvement of estrogen receptor and pCREB mediated nootropic effect of the extract.

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Acknowledgements

We are grateful to Defence Research and Development Organization, India to financially support the study financially. Authors also acknowledge Dr. Bhuvnesh Kumar, Ex-Director DIHAR for his guidance help and support for conducting the study.

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    Authors and Affiliations

    1. Defence Institute of High Altitude Research, Defence Research and Development Organisation, C/o 56 APO, Ladakh, Leh, Jammu And Kashmir, 901205, India

      Deepti Sharma, Surya Narayan Biswal, Kushal Kumar, Pushpendar Bhardwaj, Ashish Kumar, Sunil Kumar Hota & Om Prakash Chaurasia

    2. All India Institute of Medical Sciences, Sijua, Bhubaneswar, Odisha, 751019, India

      Kalpana Kumari Barhwal

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    1. Deepti Sharma
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    2. Surya Narayan Biswal
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    Deepti Sharma and Surya Narayan Biswal have contributed equally as first author.

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    11064_2017_2395_MOESM1_ESM.tif

    Table S1 and S2: shows the effect of acute and chronic (saline control animals), 300 and 2000 mg/kg of CSE extract treatment on animals body weight, food intake, water intake respectively. Data represented as mean±SEM (TIF 67 KB)

    Supplementary material 2 (TIF 68 KB)

    11064_2017_2395_MOESM3_ESM.tif

    Figure S3: Representative bright field image of Hoechst staining in CA1 region of hippocampus at different dose concentration (TIF 254 KB)

    11064_2017_2395_MOESM4_ESM.tif

    Figure S4: Representative Immunoblots showing (a) expression of ERK1/2, Elk-1 in cytosolic fraction and CREB expression in nuclear fraction. (b) Densitometric graphs depict the change in the expression of (i) ERK1/2, (ii) Elk-1 and (iii) expression of CREB following exposure to hypobaric hypoxia and administration of CSE along with ER-β antagonist PHTPP and ERK inhibitor PD98059, respectively. Data represented as mean±SEM. Bar from left to right denotes N = Vehicle treated Normoxia, NC = CSE 200 mg/kg treated normoxia, H = Vehicle treated Hypoxia and HC = CSE 200 mg/kg treated hypoxia, HC+PHTPP = CSE 200 mg/kg treated hypoxia with ERβ inhibitor PHTPP, HC+PD98059 = CSE 200 mg/kg treated hypoxia with ERK inhibitor PD98059 ‘*’ denotes p≤0.05 when compared to N, ‘#’ denotes p≤0.05 when compared to H (TIF 153 KB)

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    Sharma, D., Biswal, S.N., Kumar, K. et al. Estrogen Receptor β Mediated Neuroprotective Efficacy of Cicer microphyllum Seed Extract in Global Hypoxia. Neurochem Res 42, 3474–3489 (2017). https://doi.org/10.1007/s11064-017-2395-5

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