CHARACTERIZATION, IN VITRO ANTIOXIDANT AND HEPATOPROTECTIVE ACTIVITY OF FUNGAL ENDOPHYTIC EXTRACTS OF ANDROGRAPHIS PANICULATA LEAVES IN CCL4 INDUCED HEPATOTOXICITY

Authors

  • Smita K. Puri PG Depatment of Pharmacognosy and Phytochemistry, SET’s College of Pharmacy, S R Nagar, Dharwad 580002, Karnataka, India,
  • Prasanna V. Habbu PG Depatment of Pharmacognosy and Phytochemistry, SET’s College of Pharmacy, S R Nagar, Dharwad 580002, Karnataka, India,
  • Preeti V. Kulkarni PG Department of Pharmacology, SET’s College of Pharmacy, S R Nagar, Dharwad 580002, Karnataka, India
  • Venkatrao H. Kulkarni PG Department of Pharmacology, SET’s College of Pharmacy, S R Nagar, Dharwad 580002, Karnataka, India

DOI:

https://doi.org/10.22159/ijpps.2019v11i1.26832

Keywords:

Endophytic fungi, Hepatoprotective, Antioxidant, Andrographis paniculata, Phyllosticta sp ZLY-2010 isolate M13, Aspergillus tubingensis strain Cs72

Abstract

Objective: To isolate and characterize the endophytic fungi from the leaves of Andrographis paniculata for free radical scavenging antioxidant and hepatoprotective activity against CCl4 induced hepatotoxicity

Methods: Two fungal endophytes, APLF-3 (Andrographis paniculata leaf fungi-3) and APLF-4 (Andrographis paniculata leaf fungi-4) were isolated from leaves of Andrographis paniculata to get chloroform (A3C, A4C), ethyl acetate (A3EA, A4EA) and n butanol (A3nB, A4nB) extracts. rDNA sequencing by PCR technique was carried out for identification of APLF-3 and APLF-4. All the APLF-3 and APLF-4 extracts were assayed for in vitro free radical scavenging activity against 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical and reducing power. Then, A4EA and A4nB were screened for hepatoprotective activity against CCl4 induced hepatotoxicity at 50 mg/kg and100 mg/kg doses.

Results: The endophytic fungi, APLF-3 and APLF-4, were identified as Phyllosticta sp. ZLY-2010 isolate M13 and Aspergillus tubingensis strain Cs/7/2 respectively based on their morphological and molecular characterization. A4EA and A4nB showed significant in vitro free radical scavenging activity as compared to other extracts. A4EA and A4nB (50 mg/kg and100 mg/kg) reversed the increased serum biochemical parameters as compared to CCl4 treated group (p<0.001). A4EA and A4nB (100 mg/kg p. o) also restored the LPO, SOD and CAT levels.

Conclusion: These findings suggested that the extracts (A4EA and A4nB) obtained from endophytic fungi APLF-4 contributed towards hepatoprotective activity.

Downloads

Download data is not yet available.

References

Tian S, Yang Y, Liu K, Xiong Z, Xu L, Zhao L. Antimicrobial metabolites from a novel halophilic Actinomycete nocardiopsis terrae YIM 90022. Nat pro res 2014;28:344-6.

Egamberdieva D, Berg G, Lindstrom K, Rasanen La. Co-inoculation of Pseudomonas sp. with Rhizobium improves growth and symbiotic performance of fodder galega (Galega orientalis Lam.). Euro J of Soil Bio 2010;46:269-72.

Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Micro and Molr Bio Rev 2003;67:491–502.

Katoch M, Singh A, Singh G, Wazir P, Kumar R. Phylogeny, antimicrobial, antioxidant and enzyme-producing potential of fungal endophytes found in Viola odorata. Annals Microl 2017;67:529-40.

Swarnalatha Y, Saha B. Enhanced cytotoxic activity of endophytic bacterial extracts from Adhatoda beddomei leaves in A549 lung cancer cell lines. J Cancer Res Thera 2016;12:1284.

Fuchs B, Krischke M, Mueller MJ. Herbivore‐specific induction of defense metabolites in a grass endophyte association. Funct Ecol 2017;31:318-24.

Eidi A, Mortazavi P, Bazargan M, Zaringhalam J. Hepato-protective activity of cinnamon ethanolic extract against CCI4-induced liver injury in rats. Exp Clin Sci J 2012;11:495.

Avinash KS, Krishnamurthy YL. Cytotoxic activity of endophytic fungi HHPCYL03 isolated from Cymbopogon flexuosus nees ex steud. Int J Curr Pharm Res 2015:7:70-2.

Raval PR, Raval RM. Treatment of infective hepatitis. Where biomedicine has no answers, ayurveda has. Ancient Sci Life 2016;35:176.

Rao YK, Vimalamma G, Rao CV, Tzeng YM. Flavonoids and andrographolides from Andrographis paniculata. Phytochemistry 2004;65:2317-21.

Ghosh N, Ghosh R, Mandal V, Mandal SC. Recent advances in herbal medicine for the treatment of liver diseases. Pharma-Bio 2011;49:970-88.

Arunachalam C, Gayathri P. Studies on bioprospecting of endophytic bacteria from the medicinal plant of Andrographis paniculata for their antimicrobial activity and antibiotic susceptibility pattern. Int J Curr Pharma Res 2010;2:63-8.

Katoch M, Singh G, Sharma S, Gupta N, Sangwan PL, Saxena AK. Cytotoxic and antimicrobial activities of endophytic fungi isolated from Bacopa monnieri (L.) pennell (Scrophulariaceae). BMC Complementary Altern Med 2014;14:52.

Kumar S, Kaushik N. Endophytic fungi isolated from oilseed crop Jatropha curcas produces oil and exhibit antifungal activity. PloS One 2013;8:e56202.

Wicklow DT, Joshi BK, Gamble WR, Gloer JB, Dowd PF. Antifungal metabolites (monorden, monocillin IV, and cerebrosides) from Humicola fuscoatra Traaen NRRL 22980, a mycoparasite of Aspergillus flavus sclerotia. Appl Environ Microl 1998;64:4482-4.

Kokate CK, Purohit AP, Gokhale SB. Practical pharmacognosy. 4th Ed. Pune, Nirali Prakashan; 2005. p. 262-3.

Khandelwal KR. Practical pharmacognosy techniques and experiments. 9thEd. Pune, Nirali Prakashan; 2002. p. 149-60.

Astrid L. Preparation of lactophenol cotton blue slide mounts. Comm Eye Health 1999;12:24.

Junichi K, Makoto U, Sakae A. PCR-mediated detection of endophytic and phytopathogenic fungi from Needles of the Japanese black pine, Pinus thunbergii. Open J Forestry 2015;5:431-42.

Mathur SB, Kognsdal O. Common laboratory seed health testing methods for detecting Fungi (first Ed.) Inter seed testing association, Copenhagen, Denmark; 2003.

Saitou N, Nei M. The neighbor-joining method, a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406-25.

Kim J, Rohlf Sokal RR. The accuracy of phylogenetic estimation using the neighbor-joining method. Evolution 1993;47:1486.

Harborne AJ. Phytochemical methods a guide to modern techniques of plant analysis. Springer sci and busi media; 1998.

Wagner H, Bladt S. Plant drug analysis. a thin layer chromatography atlas. Springer Sci Business Med; 1996.

Coruh N, Celep AS, Ozgokce F. Antioxidant properties of Prangos ferulacea (L.) Lindl., Chaerophyllum macropodum boiss and Heracleum persicum desf from apiaceae family used as food in Eastern Anatolia and their inhibitory effects on glutathione-S-transferase. Food chem 2007;100:1237-42.

Halliwell B, Gutteridge JMC, Aruoma OI. The deoxyribose method, a simple ‘test tube’ assay for determination of rates constants for reactions of hydroxyl radical. Anal Biochem 1987;165:215-24.

Oyaizu M. Studies on products of the browning reaction. Antioxidative activities of products of browning reaction prepared from glucosamine. Japanese J Nutr 1986;4:307-15.

Suja SR, Latha PG, Pushpangadan P, Rajasekharan S. Evaluation of hepatoprotective effects of Helminthostachys zeylanica (L.) hook against carbon tetrachloride-induced liver damage in Wistar rats. J Ethnopharma 2004;92:61-6.

Banerjee D, Maity B, Nag SK, Bandyopadhyay SK, Chattopadhyay S. Healing potential of Picrorhiza kurroa (Scrofulariaceae) rhizomes against indomethacin-induced gastric ulceration. a mechanistic exploration. BMC Complementary Altern Med 2008;8:3.

Flohe O. Superoxide dismutase assays. In: Methods in enzmology (L Packer Ed) Academic Press: New York; 1984; 105:93-104.

Claiborne A. Handbook of methods for oxygen radical research. CRC Press; London: 1985. p. 283–4.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biochem 1951;193:265-75.

Galigher AE, Kozloff EN. In: Essentials of practical microtechnique. 2nd Edn. Lea and Febiger, Philadelphia; 1971. p. 77–9.

Deeb B, Fayez K, Gherbawy Y. Isolation and characterization of endophytic bacteria from Lectranthus tenuiflorus medicinal plant in Saudi Arabia desert and their antimicrobial activities. J Plant Inter 2013;8:56–64.

Gaiero JR, McCall CA, Thompson KA, Day NJ, Best AS, Dunfield KE. Inside the root microbiome bacterial root endophytes and plant growth promotion. Am J Bot 2013;100:1738-50.

Yang X, Zhang L, Guo B. Preliminary study of a vincristine-producing endophytic fungus isolated from leaves of Catharanthus roseus. Chintrad Herb Drug 2004;35:79-81.

Ueda JY, Tezuka Y, Banskota AH, Le Tran Q, Tran QK, Harimaya Y, et al. Antiproliferative activity of vietnamese medicinal plants. Bio Pharma Bull 2002;25:753-60.

Maiya S, Grundmann A, Li X, Li SM, Turner G. Identification of a hybrid PKS/NRPS required for pseurotin a biosynthesis in the human pathogen Aspergillus fumigatus. Chembiochem 2007; 8:1736-43.

Kour A, Shawl AS, Rehman S, Sultan P, Qazi PH, Suden P, et al. Isolation and identification of an endophytic strain of Fusarium oxysporum producing podophyllotoxin from Juniperus recurva. World J Microl Biotech 2008;24:1115-21.

University of Georgia. Plant Pathology Phyllosticta; 2015.

Wijeratne EK, Paranagama PA, Marron MT, Gunatilaka MK, Arnold AE, Gunatilaka AL. Sesquiterpene quinones and related metabolites from Phyllosticta spinarum, a fungal strain endophytic in Platycladus orientalis of the sonoran desert. J Nat Prod 2008;71:218-22.

Evidente A, Cimmino A, Andolfi A, Vurro M, Zonno MC, Cantrell CL, et al. Phyllostictines A–D, oxazatricycloalkenones produced by Phyllosticta cirsii, a potential mycoherbicide for Cirsium arvense biocontrol. Tetrahedron 2008;64:1612-9.

Glienke C, Pereira OL, Stringari D, Fabris J, Kava-Cordeiro V, Galli-Terasawa L, et al. Endophytic and pathogenic Phyllosticta species, with reference to those associated with Citrus Black Spot. Persoonia. Mol Phylogeny Evolution Fungi 2011;26:47.

Srinivasan K, Jagadish LK, Shenbhagaraman R, Muthumary J. Antioxidant activity of endophytic fungus Phyllosticta sp. isolated from Guazuma tomentosa. J Phytol 2010;2:37-41.

Oisewacz. Heinz industrial applications. New York. Springer-Verlag Berlin Heidelberg; 2002. p. 264–5.

Samson RA, Visagie CM, Houbraken J, Hong SB, Hubka V, Klaassen CH, et al. Phylogeny, identification and nomenclature of the genus Aspergillus. Stud Mycol 2014;78:141-73.

Ma YM, Li T, Ma CC. A new pyrone derivative from an endophytic Aspergillus tubingensis of Lycium ruthenicum. Nat Prod Res 2016;30:1499-503.

Huang HB, Xiao ZE, Feng XJ, Huang CH, Zhu X, Ju JH, et al. Cytotoxic Naphtho‐γ‐pyrones from the Mangrove endophytic fungus Aspergillus tubingensis (GX1‐5E). Helv Chim Acta 2011;94:1732-40.

Yadav M, Yadav A, Kumar S, Sharma D, Yadav JP. Evaluation of in vitro antimicrobial potential of endophytic fungi isolated from Eugenia Jambolana Lam. Int J Pharm Pharm Sci 2014;6:208-11.

Vipin N, Archana G, Dinesh K. Isolation and characterization of endophytic fungi from Calotropis procera for their antioxidant activity. Asian J Pharm Clin Res 2017;10:254-8.

Soare JR, Dinis TC, Cunha AP, Almeida L. Antioxidant activities of some extracts of Thymus zygis. Free Rad Res 1997;26:469-78.

De Groot H, Noll T. The crucial role of low steady state oxygen partial pressures in haloalkane free-radical-mediated lipid peroxidation. Possible implications in haloalkane liver injury. Biochem Pharma 1986;35:15-9.

Bahar E, Ara J, Alam M, Nath B, Bhowmik U, Runi N. In vitro antioxidant and thrombolytic activity of methanol extract of Sida acuta. J Pharmacogn Phytochem 2013;2:125-33.

Adewale OB, Adekeye AO, Akintayo CO, Onikanni A, Sabiu S. Carbon tetrachloride (CCl4)-induced hepatic damage in experimental sprague dawley rats. Antioxidant potential of Xylopia aethiopica. J Phytopharma 2014;3:118-23.

Elangovan NM, Dhanarajan MS, Elangovan I. Preliminary phytochemical screening and HPTLC fingerprinting profile of leaf extracts of Moringa oleifera and Phyllanthus emblica. Int Res J Pharm Biosci 2015;2:32-40.

Published

01-01-2019

How to Cite

Puri, S. K., P. V. Habbu, P. V. Kulkarni, and V. H. Kulkarni. “CHARACTERIZATION, IN VITRO ANTIOXIDANT AND HEPATOPROTECTIVE ACTIVITY OF FUNGAL ENDOPHYTIC EXTRACTS OF ANDROGRAPHIS PANICULATA LEAVES IN CCL4 INDUCED HEPATOTOXICITY”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 11, no. 1, Jan. 2019, pp. 44-54, doi:10.22159/ijpps.2019v11i1.26832.

Issue

Section

Original Article(s)