Chemical composition of Ocimum sanctum by LC-ESI–MS/MS analysis and its protective effects against smoke induced lung and neuronal tissue damage in rats

doi:10.1016/j.biopha.2017.04.011

Biomedicine & Pharmacotherapy

Volume 91, July 2017, Pages 1-12

https://doi.org/10.1016/j.biopha.2017.04.011 Get rights and content

Abstract

Smoke induced oxidative stress is known to cause various cancers and associated health problems including lung cancer. Herbal extracts have been reported as antioxidant supplements which attenuate free radical induced oxidative damage of tissues, among which Ocimum sanctum has been reported as the elixir of life due to its innumerable health benefits. In the present study, we investigated the protective effect of O. sanctum against cracker smoke induced lung and brain tissue damage. The results of the study demonstrate that O. sanctum regulates the hematological and serum biochemical parameters such as RBC, WBC, blood urea nitrogen and creatinine kinase. O. sanctum supplementation inhibited oxidative stress as analyzed by SOD, CAT enzyme levels and i-NOS, HSP-70 protein expression. O. sanctum administration also regulated neurotransmitter levels, such as serotonin, dopamine, and regulated acetylcholine esterase levels which play a vital role in neuronal function. Further O. sanctum treatment also preserved the morphology of lung and brain tissues of smoke stress induced rats as observed by histopathology and transmission electron microscope analysis. The biodistribution of O. sanctum was showed its accumulation in key tissues such as kidney, liver, lungs and heart. The LC-ESI–MS/MS analysis of O. sanctum showed the presence of polyphenols, flavonoids and fatty acids which might be responsible for the observed anti-stress effects.

Graphical abstract

Introduction

The air pollution by means of cracker smoke is commonly observed during festivals and celebrations across the globe. In India, the fireworks are generated during Diwali festival, likewise in Kerala during Vishu festival whereas in China during New Year festival and in Taiwan lantern festival. Studies reported the hazardous composition of crackers and its particulate material which pose a major risk to the humans [1], [2], [3], [4]. These studies demonstrated the presence of organic compounds such as black/aromatic carbon, hopanes, polycyclic aromatic hydrocarbons and metals such as Sr, Na, S, Mn, Al, Pb, Sr, and harmful fumes of sulfur dioxide, oxides of nitrogen and the particulate material generated by fireworks [5], [6].

Ocimum sanctum L. belongs to the family Lamiaceae and is distributed worldwide. It is widely used in Indian traditional medicines for the treatment of various ailments. The major bioactive components of O. sanctum are ursolic acid, rosmarinic acid, flavonoids and tannins, eugenol, β-caryophyllene, methyl eugenol, β-pinene and ocimumosides [7], [8]. O. sanctum possesses diverse pharmacological effects, including antimicrobial, radioprotective, immune-modulatory, nootropic, anticancer and anti-HIV activities [9], [10], [11], [12]. It is also used as diaphoretic and to treat gastric and hepatic disorders [13], [14]. Recently we have evaluated the anti-fatigue activity of ethanolic extract of O. sanctum [15].

Previously, we analyzed the in vitro neuroprotective/antioxidant effects of O. Sanctum against H₂O₂ induced oxidative stress to study its in SH-SY5Y human neuronal cells [16]. The fire cracker used in the presented study was analyzed for its metabolites by GC–MS analysis. Among the detected compounds of snake tablets phenanthrene is also reported as one of the compound present in cigarette smoke, whereas, fluoranthene is highly carcinogenic and is categorized as one among the 16 priority pollutants as recognized by the United States environmental protection agency. Some of the metabolites identified in the cracker smoke are also present in cigarette smoke, diesel exhaust, fire crackers [35]. Fireworks imposes severe occupational/health hazard particularly to respiratory system and cause asthma and common obstructive pulmonary disease (COPD) [33]. Therefore, in the present study, we aimed to investigate phytochemical constituents of O. sanctum by LC-ESI–MS/MS and to determine its in vivo anti-stress effects against cracker smoke induced oxidative stress of Wistar rats and to investigate its biodistribution.

Section snippets

Plant material

Leaf material of O. sanctum L. leaves was, purchased from the local market and identified by Dr. K. Madhava Chetty, Botanist, Department of Botany, Sri Venkateswara (S.V.) University, Tirupati, India. Voucher specimen was deposited in the herbarium (Herbarium Accession Number 1009), Department of Botany, S.V. University, Tirupati, India.

Preparation of O. sanctum extract

O. sanctum leaves were shade dried for three days and powdered. The extraction was carried out by maceration with 70% ethanol and filtered. The filtrate was

LC-ESI–MS/MS analysis of O. sanctum

The hydroalcoholic fraction of O. sanctum possesses active metabolites such as luteolin, orientin, cosmosiin, eugenol, apigenin, luteolin 5-0-beta-d-glucopyranoside, eupalitin, ocimarin, molludistin, nepetoidin, xanthomicrol, hymenoxin, carnosic acid, demethylnobiletin, salvigenin, gardenin B, palmitic acid, menthyl acetate, ursolic acid and basilimoside which were detected at positive ion mode. Further rosmarinic acid and genkwanin were detected at the negative ion mode (Figs. 1a and b and 2

Discussion

Oxidative stress is observed in neuronal disorders like Alzheimers, Parkinson’s and in ageing. The free radical mediated stress induces damage of cells among which brain is more susceptible, due to its high content of polyunsaturated fatty acids and low content of oxidative enzymes compared to other organs. The smoke generated from the vehicles, as industrial effluent, or by cigarette smoking is highly dangerous if it is inhaled due to its toxic chemical composition. In India people working in

Conclusion

In the present study, we have evaluated the anti-stress and neuroprotective effects of O. sanctum against cracker tablet smoke induced tissue damage in rats. The GC–MS analysis of cracker tablet showed the presence of toxic chemicals that might be responsible for tissue damage. The O. sanctum hydroalcoholic extract fed rats ameliorated smoke induced blood and serum biochemical alterations. The antioxidant enzyme activities were restored and lipid peroxidation, acetylcholine esterase activity

Conflicts of interest

The authors declare that there are no conflicts of interest.

Acknowledgements

The authors are highly thankful to Dr. H.V. Batra, Director, DFRL, Mysore for his constant encouragement and providing the necessary facilities throughout the study. The authors are grateful to Dr. N. Gayathri, Professor, Mrs. Hymavathi and Mr. Ravi, Neuropathology department, NIMHANS, Bangalore for TEM study. The authors also extend their gratitude to Mr. Prasanth Bitla, University of Hyderabad, Hyderabad for LC-ESI–MS/MS analysis and Mr. Ramesh Koya, Leco Instruments, Mumbai for GC–MS study.

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Chemical composition of Ocimum sanctum by LC-ESI–MS/MS analysis and its protective effects against smoke induced lung and neuronal tissue damage in rats

Abstract

Graphical abstract

Introduction

Section snippets

Plant material

Preparation of O. sanctum extract

LC-ESI–MS/MS analysis of O. sanctum

Discussion

Conclusion

Conflicts of interest

Acknowledgements

Atmos. Environ.

Atmos. Environ.

Atmos. Res.

Atmos. Environ.

J. Hazard. Mater.

J. Hazard. Mater.

Phytomedicine

J. Ethnopharmacol.

Mutat. Res.

J. Ethnopharmacol.

Pathophysiology

Neurotoxicology

Anal. Biochem.

Anal. Biochem.

Biochim. Biophys. Acta (BBA)

Arch. Biochem. Biophys.

Biochem. Pharmacol.

Kidney Int.

Int. J. Infect. Dis.

Atmos. Environ.

J. Pharmacol. Sci.