Onosma bracteata Wall. (Boraginaceae), commonly known as “gaozaban” is a highly valuable medicinal herb, useful in the treatment of body swellings, abdominal pain, eye-related problems, fever, and urinary calculi. The present study was performed to investigate the antioxidant properties of extract/fractions, viz. ethanol (Obeth) extract, hexane (Obhex) fraction, chloroform (Obcl) fraction, ethyl acetate (Obea) fraction, butanol (Obbu) fraction, and aqueous (Obaq) fraction isolated from O. bracteata. Obea fraction showed stronger free radical quenching ability in various antioxidant assays, as compared to the other fractions. Obea fraction with effective free radical-scavenging properties was further evaluated for the antiproliferative activity against human osteosarcoma MG-63, human neuroblastoma IMR-32, and human lung cancer A549 cell lines using MTT assay. Obea fraction showed strong cytotoxicity with GI50 value of 88.56, 101.61, and 112.7 μg/ml towards MG-63, IMR-32, and A549 cells respectively. Mechanistic studies revealed that Obea fraction in osteosarcoma MG-63 cells increased reactive oxygen species (ROS) level and reduced mitochondrial membrane potential. In the presence of Obea, the cells were found to be arrested in the G0/G1 phase in a dose-dependent manner which is also confirmed by the enhancement in the early apoptotic cell population in flow cytometer analysis. Western blotting demonstrated the decrease in expression of p-NFκB, COX-2, p-Akt, and Bcl-xL, whereas upregulation was observed in the expression of GSK-3β, p53, caspase-3, and caspase-9 proteins. RT-qPCR studies revealed downregulation of Bcl-2, cyclin E, CDK2, and mortalin gene expression and upregulation in the expression of p53 genes. The antioxidant and cytotoxic potential of Obea was attributed to the presence of catechin, kaempferol, onosmin A, and epicatechin, as revealed by HPLC analysis. This is the first report regarding the antiproliferative potential of O. bracteata against osteosarcoma.
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Apoptotic protease activating factor 1
Acridine orange/ethidium bromide
B cell lymphoma 2
Chinese hamster ovary
- CO2 :
Glycogen synthase kinase
Human osteosarcoma cells
Mitochondria membrane potential
Nuclear factor kappa B
Polyacrylamide gel electrophoresis
Quantitative real-time polymerase chain reaction
Reactive oxygen species
Revolutions per minute
Sodium dodecyl sulfate
Scanning electron microscopy
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The authors thankful to University Grants Commission (UGC) - Basic Scientific Research (BSR), DST-FIST (Grant No. SR/FST/LSI-691/2016 (C)) program for providing financial assistance. The authors also acknowledge UGC, New Delhi, for the instrumentation facility under UGC-DRS V (Grant No. F4-13/2015/DRS-II), CPEPA, RUSA 2.0 scheme, UPE program, and the Centre of Emerging Life Sciences, Guru Nanak Dev University, Amritsar (India), for providing the required support and facilities.
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Kumar, A., Kaur, S., Pandit, K. et al. Onosma bracteata Wall. induces G0/G1 arrest and apoptosis in MG-63 human osteosarcoma cells via ROS generation and AKT/GSK3β/cyclin E pathway. Environ Sci Pollut Res (2020). https://doi.org/10.1007/s11356-020-11466-9
- Onosma bracteata
- Cell cycle arrest