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Antioxidant Activity of Medicinal Plants

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Antioxidants are any substance that prevents or protects oxidation of other compounds. Biological antioxidants act as a defence system against harmfull free radicals produced in our body. The free radicals are atoms or molecules having unpaired valance electrons which make it very reactive. The free radicals may be derived from reactive oxygen species(ROS) or reactive nitrogen species (RNS). When free radicles are formed, they attack the adjacent stable molecules and remove electrons from them, thereby producing new free radicles. This leads to cell injury and death. Antioxidants work by scavanging and nutralizing the free radicles (Shalaby and Shanab, 2013).

A good number of studies have been made in the antioxidant activity of plants. The importance of phenols and flavonoids in the antioxidant activity of plant extracts has been emphasized by many workers. Akter et al., (2015) reported significant antioxidant and cytotoxic effect in the leaf extract of Cinnamomum tamala collected from Dhaka, Bangladesh. Hossain, (2015) also investigated the antioxidant property in the leaves of Sarcochlamys pulcherrima from Bangladesh. Ethanolic extract recorded high phenol and flavonoid content and impressive antioxidant activity. Hawari et al., (2012) compared the antioxidant activity of Ficus elastica and Ficus bengalensis. Zulfikar et al., (2011) reported significant antioxidant activity in the ethanolic extract of Ficus racemose. Prasad et al., (2009) analysed the flavonoid content and antioxidant activities of Cinnamomum species collected from South China. Out of 5 Cinnamomum species screened, Cinnamomum tamala exhibited highest superoxide anion scavenging activity.

Several reports are available on the antioxidant activity of medicinal plant extracts in India. Jasmine et al., (2015) evaluated antioxidant and anticancer property of Ficus carica fruits. Deepa et al., (2013) investigated the correlation between the polyphenols, flavonoid and tannin content with the antioxidant activity of the methanolic and aqueous extract of Cinnamomum tamala. Significant linear correlation was recorded between the phenolic compounds and the antioxidant activity of the extracts. Palanisami et al., (2011) also reported significant antioxidant and antidiabetic activity of Indian bay leaf extract to streptozotocin induced diabetic rats. Chakraborty and Das, (2010) confirmed high antioxidant property in the leaves of Cinnamomum tamala. Sirisha et al., (2010) performed a review study on the antioxidant activity of various Ficus species. The study correlated the antioxidant activity of the plants with pharmacological activities. Kapoor et al., (2009) also reported good antioxidant activity of the essential oils and oleoresins of Cinnamomum tamala. Devi et al., (2007) evaluated the antioxidant property of Cinnamomum tamala and reported a concentration dependent scavenging activity against hydroxyl and superoxide radicals.

A good number of works have also been reported from North eastern part of India. Doley et al., (2016) estimated the total phenolics, flavonoids, antioxidant activity and antimicrobial activity of methanol extract and fractions of Sarcochlamys pulcherrima collected from Dhemaji District of Assam. Highest phenol was reported in ethyl acetate fraction followed by n-butanol fraction, methanol extract, aqueous and hexane fraction respectively. Flavonoid content was also considerably high. The n-butanol fraction recorded highest antioxidant activity. Smerq and Sharma, (2011) investigated the anti-peroxidative effect of ethanolic extract of Cinnamomum tamala in rat liver homogenate using ferrous sulphate as inducer to induce lipid peroxidation. They highlighted that the flavonoid polymers in cinnamon and antioxidant activities improves the plasma fasting glucose and oxidative stress markers. Paul et al., (2010) performed a comparative study of antioxidant activity of four medicinal plants viz. Sarcochlamys pulcherrima, Gnetum gnemon, Garcinia lancifolia and Clerodendron colebrookianum in Cachar dristict of Assam. Sarcochlamys pulcherrima was reported to have highest antioxidant activity and total phenol content as compared to others.

Isolation and Characterization of Phytocompounds:

Isolation and characterization of phytocompounds of traditionally used medicinal plants is very essential to understand their possible role in preventing and curing diseases. Chromatography and spectrometry is most widely used techniques for separation and characterization of molecules. A gas chromatograph with high resolution mass spectrometer has a great ability of to analyse organic compound. It can accurately measure the mass of compounds with very high sensitivity. Darmadi et al., (2017) from Indonesia reported 10 different compounds from the methanolic extract of Cinnamomum burmanni. Azulene was the dominant compound. Hameed et al., (2016) investigated the bioactive phyocompounds from bark of Cinnamomum zeylanicum with the aid of GC MS and reported 39 different compounds. Sohilait and Kainama, (2016) carried out GC MS analysis of bark essential oils of Cinnamomum culilawa and reported 12 components. Eugenol, safrole and methyleugenol were the major components. Dong et al., (2013) identified 9 volatile compounds from the bark of Cinnamomum cassia.

Aravind et al., (2014) identified 61 individual compounds from the bark oil of Cinnamomum malabatrum. Chaudhary and Singh, (2014) performed the chemical analysis of the bioactive extracts of Cinnamomum tamala using GC MS and reported 10 compounds in the methanolic extract whereas Kumar et al., (2012) analysed the oil extracted from Cinnamomum tamala and reported 31 components from in it. Kumar et al., (2012) also compared the chemical composition of oils collected from two different regions of India, South India and North India. The GC MS analysis of the oil from North India showed 20 constituents whereas 31 constituents were reported from the oil collected from South India. Lohani et al., (2012) investigated the variability (with respect to month and tree size) in essential oils extracted from the leaves of Cinnamomum tamala from Uttarakhand Himalaya. The active principal, cinnamaldehyde was found in higher concentration in January and October. High cinnamaldehyde was reported from small size class than higher size class. Mohan et al., (20120 also reported cinnamaldehyde and cis-linalool oxide as major components of Cinnamomum tamala oil. Uma et al., (2009) identified 38 compounds from the bioactive methanolic extract of Cinnamomum zeylanicum. Baruah et al., (2007) also investigated the essential oils of Cinnamomum tamala from Jorhat, Assam. 24 compounds were reported from the leaf whereas bark extract reported 25 compounds.

Cytotoxicity of plant extracts:

The bio active components of the medicinal plants are the starting materials for new drug discovery. But these phytochemicals of the plants may also endow the plant with cytotoxic properties. Some of the toxicities which are commonly associated with the medicinal plants are allergy, gastrointestinal tract irritation, destruction of RBC, organs damage (kidney and heart) and carcinogenicity ( Nondo, et al., 2015 and IARC, 2012). Cell-based assays also remains a part of pre clinical pharmaceutical discovery. As toxicity assay gives direct impact of chemical compounds on different cell types, it is often used to screen compounds for synthesizing new drugs (Hassanein et al., (2011) & Hansen et al., (1989)). A ideal drug should be cytostatic, with negligible toxic effect to healthy cells (Radovanovic, 2015). However, the cytotoxic potential of extracts may also be exploited in preparation of anti cancerous drugs.

There are several reports on the cytotoxicity study of traditionally used medicinal plants from different parts of the world (Kuete et al., 2017; Ogbole et al., 2017; Khusbhoo et al., 2016; Neemati et al., 2013; Vijayarathna and Sasidharan, 2012; Sundaram et al., 2011; and Said et al., 2014). Rahman et al., (2013) studied the cytotoxic potential of ethanolic leaf extract of Cinnamomum tamala against brine shrimps and found it to be moderately toxic. Akter et al., also reported significant cytotoxic effect of the ethanolic leaf extract of Cinnamomum tamala. Ullah et al., (2013) evaluated the protective effect of ethanolic leaf extracts of Cinnamomum tamala against nephrotic damage in rabbits induced by gentamicin. They reported that the renal damage caused by gentamicin can be prevented when gentamicin is administered along with C tamala. In vitro cytotoxicity study of petroleum ether, methanolic and aqueous extracts was studied by Thanekar et al., (2013) against normal human cells and cancerous cells. The extracts were non toxic against normal human cells but exhibited various degree of cytotoxicity against cancerous cells. Lestari et al., (2017) evaluated the cytotoxic and anti metastatic effect of essential oils obtained from Cinnamomum burmannii toward 4T1 breast cancer cells. In their study they found that the essential oil obtained from C burmannii were potentially cytotoxic and have a tendency to inhibit 4T1 cell migration.

Computational Studies:

Traditional method of drug discovery faces a lot of challenges as it is a time consuming and labour intensive exercise which is highly expensive. These challanges can be reduced to a grater extent by using modern Bioinformatic tool which has become increasingly important in the process of designing and or discovering new drugs (Qaraghuli et al., 2017 and Leelananda & Lindert, 2016). In order to be an effective drug, a compound must be absorbed, distributed and stay in the target organ in sufficient concentration for its activity (Daina et al., 2017). Several computational tools are available to screen the drug likeness of any compound. Molecular docking studies has become one of the most powerful computational tool in drug designing by studying protein-ligand interactions. AutoDock suite is one of the widely used tool in drug research and it include several versions like AutoDock Vina, AutoDock, Raccoon 2, AutoDock Tools and Autodock ligands (Forly et al., 2016). AutoDock vina is a recently developed tool, highly optimized to perform docking experiments. The use of docking in the field of drug designing have witnessed significant rise in the last 25 years (Ramirez and Caballero, 2016). There are numerous reports of in silico studies of compounds isolated from plants or synthethic compounds against known protein targrts. Varalakshmi et al., (2017) performed molecular docking of Procyanidin B2 isolated from the methanolic extract of bark of Cinnamomum zeylenicum against transcription factor Nf-kB. The ligannd was found to be a inhibitor of the transcription factor. Lestari et al., (2017) studied docking of cinnamaldehyde isolated from Cinnamomum burmannii barks against target protein, MMP-2 and MMP-9 and predicted molecular interaction. Stronger binding of the ligands with the targets were reportes. Compounds isolated from petroleum ether fraction of Cinnamomum tamal were docked by Thanekar, et al., 2016 against the enzyme topoisomerase I. Eugenol and α-caryophyllene were predicted to show inhibitory activity against topoisomerase I. Sumiwi et al., (2012) studied molecular docking of compounds isolated from essential oils of Cinnamomum sintoc against COX-2 and found 4- terpinol to be the most significant inhibitor of the protein. Sivakumari et al., (2010) studied in silico inhibiting efficiency of cinnamaldehyde, cinnamic acid and cinnamal alcohol (isolated from Cinnamomum cassia) against aldose reductase enzyme which is associated with diabetic ratinopathy. Cinnamaldehyde was found to be most active inhibitor as compared to others.

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