Journal of Applied Biology & Biotechnology Vol. 9(1), pp. 83-87, Jan-Feb, 2021 Available online at http://www.jabonline.in DOI: 10.7324/JABB.2021.9111 Antimicrobial, anti-inflammatory, and anticancer activities of leaves extracts of Filicium decipiens Atiyaparveen I Basarikatti1, Sanjay Mishra2,3, Vijaykumar Uppar1, Basavaraj Padmashali1* 1Department of Chemistry, Rani Channamma University, Belagavi, Karnataka, India. 2KAHERs Dr. Prabhakar Kore Basic Science Research Centre, KLE Academy of Higher Education and Research, Belagavi, Karnataka, India. 3KAHER’s College of Pharmacy, KLE Academy of Higher Education and Research, Nehru Nagar, Belagavi, Karnataka, India. ARTICLE INFO ABSTRACT Article history: Filicium decipiens (fern-tree) belongs to the family Sapindaceae. It grows in Asia and Africa region and is traditionally Received on: July 14, 2020 used for diabetes treatment in India. The plant has been documented to have molluscicidal, diabetic nephropathy, Accepted on: October 22, 2020 Available online: January 17, 2021 and anti-inflammatory properties. In the present study, experimental data on different leaves extracts of F. decipiens were collected for their preliminary phytochemical investigations along with antimicrobial, anti-inflammatory, and anticancer activities using in-vitro assays. The phytochemical investigations display the existence of alkaloids, Key words: Filicium decipiens, tannins, flavonoids, saponins, glycosides, steroids, triterpenoids, and carbohydrates in the extracts prepared using Anticancer, three different solvents. Antimicrobial property was established against six bacterial and fungal strains. Petroleum Antimicrobial and anti-inflammatory ether extract exhibited higher antimicrobial activity as compared to chloroform and water extract. Anti-inflammatory assay was performed on RAW264.7 murine macrophage cells and petroleum ether extract exhibited reduced nitrite oxide production having IC50 value 493.7 µg/ml. Cytotoxicity was determined against MCF-7 (Breast) and A-549 (Lung) cancer cells and paclitaxel was used as a standard drug, whereas cytocompatibility was assessed against mouse fibroblast cells (L929). Different sensitivities were observed against different study cell lines in a dose- dependent manner. These study findings may provide biological evidence for the application of Filicium decipiens extract. 1. INTRODUCTION an essential role in cancer management and indeed over the last half-century many therapeutic plant/s along with their secondary Plants have been used as a source of raw material for remedies metabolites have been employed [4]. Rationale use of medicinal since ancient practice and several of them are globally recognized plants is an effective approach in the chemotherapeutic management as medicine [1]. They contain biologically active constituents to of cancer and is one of the major parts of treatment for several years protect human beings and such constituents act as protective agents as many medicinal plants with anticancer activity are previously against several human ailments steps such as mortal carcinogenesis, documented in literature [5]. As the attention in simple and organic performing origination, development phases, or terminating the lifestyle develops, the awareness in plant-based drugs also rises. DNA destructing cell mutations. These medicinal plant sources could In addition to this, side effects and interaction possibilities are probably be used to offer more efficacious anticancer agents [2]. Such boundaries in synthetically developed anticancer medicines. Hence, properties are well co-related previously to the existence of certain plant/s has been studied globally to explore them as possible sources phytochemicals. Hence, medicinal plants play an active role in cancer of anticancer agents [6]. prophylaxis and management [3]. Filicium decipiens is a tropical fern-like leaf tree which grows The identification and assessment of biologically active plant in Asia and Africa. The plant is usually found in high altitude extract/s have been of abundant importance to biomedical scientists regions up to 1000 m [7]. It is an evergreen tree found abundantly in the exploration of newer and safer drug toward the disease in peninsular India. The leaves are compound and very large. management. Cancer is a second most deadly disease in the world Each leaf consists of 12–16 leaflets, each leaflet is 4–6 inches and currently many clinically effective regimens are in practice. in length and relatively narrow [8]. The flowers are very small The plant-based drug discovery and development have displayed and white in color and the tree produces both male and female flowers. The fruits are olive size, drupe and dark in color, and *Corresponding Author form in clusters. The stem bark is black in color [9]. Ethanolic Basavaraj Padmashali, Department of Chemistry, Rani Channamma extract of F. decipiens has been previously documented for anti- University, Belagavi - 591 156, Karnataka, India. inflammatory property and sitosterol has been recognized from E-mail: [email protected] the methanolic extract of F. decipiens leaves. Furthermore, four © 2021 Basarikatti, et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-ShareAlike Unported License (http://creativecommons.org/licenses/by-nc-sa/3.0/). 84 Basarikatti, et al.: Journal of Applied Biology & Biotechnology 2021;9(1):83-87 new saponins were identified and isolated from of stem bark of broth and the extracts of F. decipiens. The tubes were incubated F. decipiens [10]. In view of the above effects, this study aimed aerobically at 37°C for 24–48 h [12]. The least concentration (higher to evaluate preliminary phytochemical investigations and in-vitro dilution) of the extract does not produce any growth (no turbidity) efficacy for antimicrobial, anti-inflammatory, and anticancer in the 24 h. They are compared with control tubes and are recorded potential of different leaf extracts of F. decipiens. as MIC. The value of MBC was decided by sub-culturing test dilution (MIC) 2. MATERIALS AND METHODS against newly prepared BHI agar plates. The plates were incubated 2.1. Chemicals and Reagents for 24 h at 37°C. The maximum dilution displayed no single bacterial The solvents used for extraction were obtained from Merck colony on the BHI agar plates which were recorded as MBC. While Limited, Mumbai, India. Dulbecco’s Modified Eagle’s Medium the minimum concentration that stops fungal growth after aerobic (DMEM), lipopolysaccharide (LPS), aminoguanidine (AG), 3-(4, incubation was recorded as MFC. 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT), brain heart infusion (BHI) agar, nutrient broth, fetal bovine serum 2.5. Anti-inflammatory Activity (FBS), penicillin, and streptomycin were obtained from HiMedia Pvt. 2.5.1. Cell culture of RAW 264.7 Ltd., Mumbai, India. RAW 264.7 (Murine macrophage cells) was cultured in DMEM with 2 mM L-glutamine having 10% HI-FBS, 100 IU/ml penicillin, and 2.2. Plant Material 100 µg/ml streptomycin in a humidified atmosphere at 37°C in the The plant was identified and authenticated from ICMR-National presence of 5% CO2 [13]. Institute of Traditional Medicine. The plant specimen was preserved 2.5.2. Cell viability – MTT assay in the herbarium (RMRC-1388). F. decipiens (Sapindaceae) leaves The murine macrophage cells were grown at a density of were collected from the campus of Sahyadri Science College Shimoga 2 × 105 cells/ml in 96 well plates with the extracts of F. decipiens (Karnataka). The collected leaves were washed with the help of water (25–800 µl/ml). 1 µg/ml LPS of cells was incubated. Two hundred and were shade dried at room temperature (32 ± 2°C). The leaves micrometer AG along with 1 µg/ml LPS was considered as control were grinded and the powdered mass was stored for the subsequent of decrease in nitrite oxide (NO) formation. For nitrite quantity, extraction procedure. the culture supernatant has been removed. Five microliter of MTT 2.2.1. Extraction solution was added and the plates were incubated at 37°C for 4 h. The leaves powder (200 g) was extracted individually using petroleum To thaw the formazan crystal, the solution of MTT was detached ether, chloroform, and water (1500 ml) in a Soxhlet assembly for and dimethyl sulfoxide (DMSO) was added to the plate. Incubation 48 h followed by filtration and evaporation process using a rotary for 10 min at room temperature absorbance was recorded at 540 nm evaporator at 35–40°C to obtain petroleum ether (FDP), chloroform using ELISA plate reader [14,15]. (FDC), and water (FDH) extracts separately. 2.5.3. Nitrite oxide determination NO was determined by evaluating the nitrite with the help of Griess 2.3. Phytochemical Analysis reagent. With the equal volume of cell culture was mixed with Griess The leaves powder of F. decipiens was extracted with petroleum reagent [16] followed by incubation for 10 min, diazo dye was formed. ether, chloroform, and water as solvents to get polar and non-polar Absorption of the formed diazo dye was assessed at 540 nm. For the components. Different phytochemical tests have been assessed for evaluation of nitrite concentration, it was determined by comparing alkaloids, flavonoids, glycosides, phenol, saponins, steroids tannins, with a sodium nitrite standard calibration curve. proteins, and carbohydrates by following the previously reported procedure of Debela. [11]. 2.6. Cytotoxic Screening 2.6.1. Cell culture and dilution of extracts