Abstract
In this study, we assessed the potential of aqueous extract (CSEaq) of Cuminum cyminum L. (cumin) seeds in protecting WRL-68 cells from hexavalent chromium [Cr(VI)]-induced oxidative injury. Cells exposed to Cr(VI) (10 μM CrO3) for 24 h demonstrated a twofold increase in ROS, which, in turn, led to extensive oxidative stress, consequently causing colossal decline in cell viability (by 58.82 ± 9.79%) and proliferation (as was evident from a reduced expression of Ki-67, a proliferation marker). Immunofluorescence studies showed that Cr(VI) diminished the expressions of mTOR and survivin in WRL-68 cells. It also led to a substantial elevation of BECN1 expression, which suggested autophagy. Overall, our results indicated that 24 h exposure of WRL-68 cells to Cr(VI) caused oxidative stress-induced autophagic cell death. CSEaq was found to protect WRL-68 cells from the same fate by refurbishing their viability and proliferation in a dose-dependent manner. The extract reduced ROS in these cells, which consequently decreased the degree of autophagic cell death by restoring expressions of mTOR, survivin and BECN1 to their respective normal levels. Biochemical assays revealed that CSEaq is rich in phenolic constituents. Total phenolic content of CSEaq demonstrated positive correlations with (i) its antioxidant potential, (ii) its alleviation of cellular oxidative stress and (iii) its cytoprotective efficacy in Cr(VI)-treated WRL-68 cells. We also identified the major phenolic constituents of CSEaq. Our study suggested that polyphenols in CSEaq might be responsible for protecting WRL-68 cells from Cr(VI)-governed oxidative assault that would have otherwise led to survivin/mTOR-mediated autophagic death.
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Abbreviations
- AGE:
-
Advanced glycation end product
- BECN1:
-
Beclin-1
- Cr(VI):
-
Hexavalent chromium
- CrO3 :
-
Chromium trioxide
- CSEaq :
-
Aqueous extract of cumin seeds
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- DCFDA:
-
2′,7′-Dichlorofluorescin diacetate
- DMSO:
-
Dimethyl sulphoxide
- DPPH· :
-
2,2-Diphenyl-1-picrylhydrazyl radical
- DSA:
-
DPPH· scavenging activity
- GAE:
-
Gallic acid equivalence
- HPLC:
-
High-performance liquid chromatography
- MEM:
-
Minimum essential medium Eagle
- mTOR:
-
Mechanistic target of rapamycin
- mTORC1:
-
MTOR complex 1
- MTT:
-
3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide
- PARP-1:
-
Poly(ADP-ribose) polymerase-1
- PBS:
-
Phosphate buffered saline
- PI3K-III:
-
Class III phosphatidylinositol 3-kinase complex
- PI3P:
-
Phosphatidylinositol-3-phosphate
- ROS:
-
Reactive oxygen species
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Acknowledgements
We thank Dr. S. Nagarajan, Department of Chemistry, National Institute of Technology Warangal, India, for his support. We also thank Ms. K. Priyadharshini for her help with the experiments.
Funding
This study was funded by the Grants YSS/2014/000139 (GG), YSS/2015/000025 (DB-G), ECR/2016/001856/ES (BNVH) and SR/FST/ETI-331/2013 (SASTRA) from the Department of Science and Technology Science (DST), Government of India; and BT/PR22434/MED/30/1901/2017 (GG and DB-G) from the Department of Biotechnology (DBT), Government of India.
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Idea conception and experiment designs by: GG and DB-G; Experiments performed by: RM, JP and BNVH; Data analyzed by: GG, DB-G, RM and JP; Paper written by: GG and DB-G.
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Mahalakshmi, R., Priyanga, J., Vedha Hari, B.N. et al. Hexavalent chromium-induced autophagic death of WRL-68 cells is mitigated by aqueous extract of Cuminum cyminum L. seeds. 3 Biotech 10, 191 (2020). https://doi.org/10.1007/s13205-020-02184-7
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DOI: https://doi.org/10.1007/s13205-020-02184-7