In Silico Target Identification and Validation for Antioxidant and Anti-inflammatory Activity of Selective Phytochemicals

Abstract Phytochemicals present in plant extract include a number of biological active compounds which have shown promising antioxidant and anti-inflammatory activities in many animal studies. Present knowledge about the biochemical interactions of these compounds present in phytochemical extracts and target enzymes or proteins responsible for antioxidant and anti-inflammatory activity is limited. Present work is an attempt to identify and validate possible biological targets as enzymes or proteins involved in these targeted studies using molecular docking as computational method. IMPPAT: Indian Medicinal Plants, Photochemistry and Therapeutics (a curated database) has been used to retrieve various phytochemicals derived from selected plants which includes Carica papaya, Citrus limon, Curcuma longa, Dalbergia sissoo and Punica granatum. These phytochemicals are further evaluated using molecular docking against three enzymes involved in antioxidant activity which includes Superoxide dismutase (SOD), Glutathione peroxidase (GPX) and Catalase (CAT). Cyclooxigenase-2 (COX-2) has been tested for anti-inflammatory activity of these phytochemicals. Gliadin (Triticum aestivum), Tea Extract (Punica granatum), Hesperidin (Citrus limon), Terrestribisamide (Triticum aestivum), Vitamin P (Carica papaya) and 1,2,6-Trigalloylglucose (Punica granatum) are few phytochemicals which has shown promising binding affinities towards target proteins or enzymes Superoxide dismutase (SOD), Glutathione peroxidase (GPX) and Catalase (CAT) and cyclooxigenase-2 (COX-2).

Isolation and evaluation of biological efficacy of quercetol in human hepatic carcinoma cells.

Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human and animal health. The current work aimed to test cytotoxic and apoptotic effects of quercetol on HepG2 cells. Quercetol was isolated from Ocimum sanctum and characterized by gas chromatography-tandom mass spectrometry (GC-MS/MS), nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. Quercetol (50-600 µg/mL) was examined for cytotoxic activity by tetrazolium salt and neutral red uptake tests and comet assay for genotoxicity, using HepG2 cells, over 24 hours. Data from 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and neutral red uptake tests demonstrated quercetol-induced cytotoxicity in HepG2 cells in a concentration-dependent manner. With 4',6-diamidino-2-phenylindole staining, a significant induction of chromosomal condensation was observed at 300 µg/mL of quercetol. DNA fragmentation analysis showed that quercetol produced cell death in HepG2 cells in a concentration-dependent manner. Thus, our study suggests that an environmentally relevant concentration of quercetol, which was a chemically standardized extract from O. sanctum, induced cell death and DNA damage in HepG2 cells.

Isolation and evaluation of anticancer efficacy of stigmasterol in a mouse model of DMBA-induced skin carcinoma.

Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced skin cancer was investigated in Swiss albino mice. Skin tumors were induced by topical application of DMBA and promoted by croton oil. To assess the chemopreventive potential of stigmasterol, it was orally administered at a concentration of 200 mg/kg and 400 mg/kg three times weekly for 16 weeks. Reduction in tumor size and cumulative number of papillomas were seen as a result of treatment with stigmasterol. The average latency period was significantly increased as compared with the carcinogen-treated control. Stigmasterol induced a significant decrease in the activity of serum enzymes, such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin as compared with the control. Stigmasterol significantly increased glutathione, superoxide dismutase, and catalase as compared with the control. Elevated levels of lipid peroxide and DNA damage in the control group were significantly inhibited by administration of stigmasterol. From the present study, it can be inferred that stigmasterol has chemopreventive activity in an experimental model of cancer. This chemopreventive activity may be linked to the oxidative stress of stigmasterol. The antigenotoxic properties of stigmasterol are also likely to contribute to its chemopreventive action.

Quercetin attenuates the development of 7, 12-dimethyl benz (a) anthracene (DMBA) and croton oil-induced skin cancer in mice.

To evaluate the chemopreventive potential of quercetin in an experimental skin carcinogenesis mouse model. Skin tumor was induced by topical application of 7, 12-dimethyl Benz (a) anthracene (DMBA) and Croton oil in Swiss albino mouse. Quercetin was orally administered at a concentration of 200 mg/kg and 400 mg/kg body weight daily for 16 weeks in mouse to evaluate chemopreventive potential. Skin cancer was assessed by histopathological analysis. We found that quercetin reduced the tumor size and the cumulative number of papillomas. The mean latent period was significantly increased as compared to carcinogen treated controls. Quercetin significantly decreased the serum levels of glutamate oxalate transaminase, glutamate pyruvate transaminase, alkaline phosphatase and bilirubin. It significantly increased the levels of glutathione, superoxide dismutase and catalase. The elevated level of lipid peroxides in the control group was significantly inhibited by quercetin. Futhermore, DNA damage was significantly decreased in quercetin treated mice as compared to DMBA and croton oil treated mice. The results suggest that quercetin exerts chemopreventive effect on DMBA and croton oil induced skin cancer in mice by increasing antioxidant activities.

Extraction optimization of Tinospora cordifolia and assessment of the anticancer activity of its alkaloid palmatine.

OBJECTIVE: To optimize the conditions for the extraction of alkaloid palmatine from Tinospora cordifolia by using response surface methodology (RSM) and study its anticancerous property against 7,12-dimethylbenz(a)anthracene (DMBA) induced skin carcinogenesis in Swiss albino mice. METHODS: The effect of three independent variables, namely, extraction temperature, time, and cycles was investigated by using central composite design. A single topical application of DMBA (100 µg/100 µL of acetone), followed 2 weeks later by repeated application of croton oil (1% in acetone three times a week) for 16 weeks, exhibited 100 percent tumor incidence (Group 2). RESULTS: The highest yield of alkaloid from Tinospora cordifolia could be achieved at 16 hours of extraction time under 40°C with 4 extraction cycles. Alkaloid administration significantly decreases tumor size, number, and the activity of serum enzyme when compared with the control (Group 2). In addition, depleted levels of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase and increased DNA damage were restored in palmatine treated groups. CONCLUSION: The data of the present study clearly indicate the anticancer potential of palmatine alkaloid in DMBA induced skin cancer model in mice.

In silico identification of drug targets for antifertility from natural products by differential reaction content analysis of metabolic pathways.

BACKGROUND: One of the major concerns of governments of developing and developed countries is to have a check on their population increase. Realising the importance of avoiding the harmful effects of synthetic compounds, scientists and researchers throughout the world are cooperating in efforts to design new and effective contraceptives from compounds of plant origin. METHODS: In this paper, we compared 11 plant species by analysing compounds showing antifertility properties with respect to the metabolic pathways involved. The Kyoto Encyclopedia of Genes and Genome pathway database was the source of metabolic pathway information. Protein sequences and classification numbers of unique enzymes exclusively present in certain species were identified using the Expert Protein Analysis System. RESULTS: Two enzymes, namely, L-aspartate dehydrogenase (EC no. 1.4.1.21) and trans-hexaprenyltranstransferase (EC no. 2.5.1.30), were identified as novel drug targets from the metabolic pathway analysis. Validation of the essential proteins identified through metabolic pathway comparison was done based on the literature information. CONCLUSION: The in silico analysis resulted in identification of 2 enzymes that are predicted to be the targets for putative antifertility drug. These enzymes can further be modelled to obtain their 3-dimensional structures with the help of various protein structure modelling softwares.

Evaluation of uptake rate of heavy metals by Eichhornia crassipes and Hydrilla verticillata.

Lakes, ponds, and streams are the sources of surface water, which anchorage the survival of aquatic life flora and fauna and maintain ecological balance. Due to urbanization, population explosion, and industrialization, these natural sources are getting polluted. Present paper is an attempt to evaluate the uptake rate of heavy metals namely lead (Pb), zinc (Zn), iron (Fe), and chromium (Cr) by the macrophytes. The two macrophytes taken for the study are Eichhornia crassipes and Hydrilla verticillata. Both macrophytes have the capacity to absorb heavy metals from contaminated water. The present experimental study was conducted to compare and identify their potential to improve the water quality by removing the heavy metals. The paper critically evaluates the water-purifying capacity of submerged macrophyte (H. verticillata) and free-floating macrophyte (E. crassipes). It also evaluates the extent up to which heavy metal can be removed by macrophyte in a given period of time.

Water quality improvement through macrophytes--a review.

Increasing urbanization, industrialization and over population is leading to the degradation of the environment. The main hazardous contents of the water pollution are heavy metals etc. Water bodies are the main targets for disposing the pollutants directly or indirectly. They are again at the receiving end as the storm water, residential and commercial waste is disposed into it. The prevailing purification technologies used to remove the contaminants are too costly and sometimes non-eco friendly also. Therefore, the research is oriented towards low cost and eco friendly technology for water purification, which will be beneficial for community. The present paper is a comprehensive review of approximately 38 literature sources. The paper discusses the potential of different aquatic plants (macrophytes) in purifying water and wastewater. Experimental work was developed to test the hypothesis that nutrient enrichment enhances metal tolerance of relative macrophyte.

Statistical evaluation of hydrobiological parameters of Narmada River water at Hoshangabad City, India.

Narmada is considered to be the lifeline of the state of Madhya Pradesh in Central India. The Narmada water is used for bathing, drinking, irrigation and industrial purposes. The city sewage and industrial effluent from Security paper mill at Hoshangabad drains in the Narmada River and pollutes the water quality. Urban sewage enters into Narmada through main nallas. River water quality at Hoshangabad has become a matter of concern due to continuous changing environment and increasing social and industrial activity that influence the water quality directly or indirectly. The present investigation is undertaken to study the effect of domestic sewage and effluent from Security paper Mill on the water quality and ecology of river Narmada at Hoshangabad. The study is carried on at four sites along with the bank of river Narmada. Water samples from four stations were collected, out of which three main sewage mixing points of the city and one fresh water site are taken into account. The samples collected were analyzed, as per standard methods parameters such as Temperature, pH, were measured in-situ. The statistical evaluations were also made. The result showed increase in BOD, Nitrates, Phosphates and Total Coliforms, No. of phytoplanktons. The results revealed that most of the water samples were below or out of limited; according to the WHO, BIS standards.

Multivariate analysis of drinking water quality parameters in Bhopal, India.

Pollution of water bodies is one of the areas of major concern to environmentalists. Water quality is an index of health and well being of a society. Industrialization, urbanization and modern agriculture practices have direct impact on the water resources. These factors influence the water resources quantitatively and qualitatively. The study area selected were the Upper lake and Kolar reservoir of Bhopal, the state capital of Madhya Pradesh, India. The Upper lake and Kolar reservoir both are the important sources of potable water supply for the Bhopal city. The physico-chemical parameters like temperature, pH, turbidity, total hardness, alkalinity, BOD, COD, Chloride, nitrate and phosphate were studied to ascertain the drinking water quality.

An innovative technique for lake management with reference to aeration unit installed at lower lake, Bhopal, India.

Water is key element in human life. All forms of life upon the earth depend upon water for their mere existence. Life & water may be aptly said to be two facets of the same coin. Most of the water bodies are getting polluted due to domestic waste, sewage, industrial waste and agricultural effluent. The present study is designed to ascertain the effectiveness of artificial aeration cum Ozonizer unit installed at Lower Lake, Bhopal for assessment of water quality. Various physico- chemical parameters like pH, Dissolved oxygen, Biochemical Oxygen demand, Chemical oxygen demand, nitrate, phosphate and bacteriological status were studied to assess the extent of deterioration in water quality of Lower lake and at the same time to assess the performance of the dual aeration system in improvement of water quality.

An effective means of biofiltration of heavy metal contaminated water bodies using aquatic weed Eichhornia crassipes.

Various aquatic plant species are known to accumulate heavy metals through the process of bioaccumulation. World's most troublesome aquatic weed water hyacinth (Eichhornia crassipes) has been studied for its tendency to bio-accumulate and bio-magnify the heavy metal contaminants present in water bodies. The chemical investigation of plant parts has shown that it accumulates heavy metals like lead (Pb), chromium (Cr), zinc (Zn), manganese (Mn) and copper (Cu) to a large extent. Of all the heavy metals studied Pb, Zn and Mn tend to show greater affinity towards bioaccumulation. The higher concentration of metal in the aquatic weed signifies the biomagnification that lead to filtration of metallic ions from polluted water. The concept that E. crassipes can be used as a natural aquatic treatment system in the uptake of heavy metals is explored.