• Sudarahana Mysore Shankarsingh professor of botany, University of Mysore India .
  • Girish Hodiyala Vasanaika faculty of botany, university of mysore India .


Leaf, Leaf callus, Alcohol, Hepatoprotection, HepG2 cell line, MTT assay


The study was carried out to evaluate the hepatoprotective activity of leaf and leaf callus extracts of Anisochilus carnosus (L) Wall. against alcohol induced toxicity using HepG2 cell line. Leaf explants were cultured on Murashige and Skoog solid medium supplemented with different growth regulators. Prior to the determination of hepatoprotective property leaf and leaf callus extracts were subjected to the toxic dose study. The degree of hepatoprotection of extracts was determined by measuring cell viability percentage by MTT assay. The preliminary phytochemical analysis of leaf and leaf callus was carried out by qualitative analysis. Maximum percentage of callus formation (98%) was obtained in MS medium fortified with 3 mg/l 2,4-D. HepG2 cells were pretreated with the different concentrations (below toxic dose) of leaf and leaf callus extracts for 72 hours followed by alcohol intoxication. Results revealed that ethanolic leaf extract pretreated HepG2 cells show 94% cell viability compared to the standard silymarin pretreated HepG2 cells which showed 81% cell viability. Leaf callus extracts also exhibited significant hepatoprotective activity where ethanolic callus extract pretreated HepG2 cells showed 86% viability after intoxication with alcohol. Results revealed that HepG2 cell viability percentage is dose dependent. Phytochemical studies revealed the presence of different secondary metabolites in leaf and leaf callus extracts. The bio-efficacy study confirms the presence of secondary metabolites of hepatoprotective nature in leaf and leaf callus of A. carnosus.


Johnson M, Wesely EG, Kavitha MS, Uma V. Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L. Asian Pacific journal of tropical medicine. 2011;4(3):196-200.

Ramoji A, Kuppast IJ, Mankani KL. Protective effect of Paracalyx scariosa against ethanol induced hepatic toxicity in BRL 3A cell lines. International Research Journal of Pharmacy. 20145;(6):495-501.

Neuman MG. Cytokines - central factors in alcoholic liver disease. Alcohol Research Health. 2003;27(10):307-316.

Kumar V, Cotran R, Robbins S. Robbins basic pathology, 7th Ed. Philadephlia: W.B. Saunders; 2003.

Wu D, Caderbaum AI. Ethanol induced apoptosis to stable HepG2 cell line expressing human cytochrome P-4502E1. Alcoholism: Clinical and Experimental Research. 1993;23(9):67-76.

Panda N, Banerjee S, Mandal NB, Sahu NP. Pragnane glycoscides. Natural Product Communications. 2006;7(1):665-669.

Trease GE, Evans WC. Pharmacognosy. London: Balliere Tindall Press; 1983:56-57.

Kamble SY, More TN, Patil SR, Pawar SG, Bindurani R, Bodhankar SL. Plants used by the tribes ofNorthwest Maharashtra for the treatment of gastrointestinal disorders. Indian Journal of Traditional Knowledge. 2008;7(2):321-325.

Subramanian SS, Nair AGR. Flavonoids of the leaves of Mentha spicata and Anisochilus carnosus. Phytochemistry. 1972;11(1):452-453.

Senatore F, Francesca L, Venza F, Bruno M, Napolitano F. Composition and antibacterial activity of the essential oil of Anisochilus carnosus- a Tamil plant acclimatized in Sicily. Flavour and Fragrance Journal. 2003;18(8):202-204.

Grover JK, Adiga G, Vata V, Rathi SS. Extracts of Anisochilus carnosus prevent development of experimental inflammation. Journal of Ethanopharmacy. 2001;7(8):159-164.

Mohammed A, Kumar RJ, Santosh HY, Nagashruthi MH. Antiulcer activity of Anisochilus carnosus leaf extracts in pylorus ligation rats. Indian Drugs. 2008;45(12): 979-983.

Kulandhaivel M, Sooraj SN, Nathiya K, Palaniswamy M. Antifungal activity and phytochemical screening of Anisochilus carnosus and Melaleuca alternifolia against opportunistic pathogen Candida albicans. International Journal of Pharmaceutical and Biological Archives. 2011;2(6):1668-1670.

Muthuraman MS, Santharam L, Ariraman S, Pemaiah B. Studies on anticancer and antimicrobial efficacy of Anisochilus carnosus extracts. International Journal of Pharmacy and Pharmaceutical Sciences. 2012;4(2):132-135.

Harborne JB. Phytochemical Methods-A guide to modern techniques of plant analysis. Chapman and Hall, London. 1998:182-190.

Escara JF, Hutton RJ. Thermal stability and renaturation of DNA in dimethyl sulfoxide solutions: acceleration of the renaturation rate. Biopolymers. 1980;19(7):1315-1327.

Thabrew M, Hughes RD, Mcfarlane IG. Screening of hepatoprotective plant components using a HepG2 cell cytotoxicity assay. Journal of Pharmacy and Pharmacology. 1997;49(11):1132-1135.

Oh H, Lee HS, Kim T, Chai KY, Chung HT, Kwon TO, Yun YG. Furocoumarins from Angelica dahurica with hepatoprotective activity on tacrine-induced cytotoxicity in Hep G2 cells. Planta Medica. 2002;68(05):463-464.

Sharma A, Gangwar M, Kumar D, Nath G, Sinha AS, Tripathi Y. Phytochemical characterization, antimicrobial activity and reducing potential of seed oil, latex, machine oil and presscake of Jatropha curcas. Avicenna Journal of Phytomedicine. 2016;6(4): 366-375.

Rout GR, Saxena C, Samantaray C, Das P. Rapid plant regeneration from callus cultures of Plumbago zeylanica. Plant Cell Tissue and Organ Culture. 1999;56:47-51

Jain SC, Jian R, Vlietinck JA. 2004. In vitro and in vivo antimicrobial efficacy of Mimosa hamate. Indian Journal of Biotechnology, 3; 271-273

Shariff N, Sudarshana MS, Umesha S, Hariprasad P. Antimicrobial activity of Rauvolfia tetraphylla and Physalis minima leaf and leaf callus extracts. African Journal of Biotechnology. 2006;5:946-950

Manickam VS, Elangomadhavan R, Antonisamy R. Regeneration of Indian ginseng plantlets from stem callus. Plant Cell Tissue and Organ Culture. 2000;14:55-58

Ibrahim M, Khaja NM, Aara A, Khan AA, Habeeb AM, Devi PY, Narasu LM, Habibullah MC. Hepatoprotective activity of Sapindus mukorossi and Rheum emodi extracts: In vitro and in vivo studies. World Journal of Gastroenterology. 2008;14(6):2566-2571.

Kai OL. Alcoholic liver disease: Pathobiological aspects. Journal of Hepatology. 1995;23(4), 7-15

Arulkumaran KSG, Rajasekaran A, Ramasamy R, Jegadeesan M, Kavimani S, Somasundaram S. Cassia roxburghii seeds protect liver against toxic effects of ethanol and carbontetrachloride in rats. International Journal of Pharma Tech Research. 2009;1(2):273-246.

Lee HB, Huang YY, Wu CS. Hepatoprotective activity of fresh Polygonum multiflorum against HepG2 hepatocarcinoma cell proliferation. Journal of Food and Drug Analysis, 2011;19(1):26-32.

Sivaraj A, Vinothkumar P, Sathiyaraj K, Sundaresan S, Devi K, Senthilkumar B. Hepatoprotective potential of Andrographis paniculata aqueous leaf extract on ethanol induced liver toxicity in albino rats. Journal of Applied Pharmaceutical Science. 2011;1(6):24-28.

Asha TNK, Londonkar RL, Nayaka HB. Cytotoxicity and hepatoprotective attributes of methanolic extract of Rumex vesicarius L. Biological Research. 2015;48(1):1-9.

Kanchana G, Jayapriya K. Antioxidant effect of Livomap, A polyherbal formulation on ethanol induced hepatotoxicity in albino Wistar rats. Journal of Applied Pharmaceutical Science. 2013;3(3):52-56.

Babbar, N, Oberoi HS, Uppal DS, Patil RT. Total phenolic content and antioxidant capacity of extracts obtained from six important fruit residues. Food Research International. 2011;44(1):391-396.

Kamali M, Khosroyar S, Kamali H, Ahmadzadeh Sani T, Mohammadi A. Phytochemical screening and evaluation of antioxidant activities of Dracocephalum kotschyi and determination of its luteolin content. Avicenna Journal of Phytomedicine. 2016;6(4):425-433.

Santhosh S, Sini KT, Anandan R, Mathew PT. Hepatoprotective activity of chitosan against isonaizid and rifampicin induced toxicity in experimental rats. European Journal of Pharmacology. 2007;57(2),:69-73.

Kamel HH, Azza H, Rehman AE, Walaa MSA, Amira HM . Protective effect of some antioxidants against CCl4 induced toxicity in liver cells from BRL3A cell line. Journal of American Science. 2010;6(10):992-1003.

Bhagyashree RP, Sanjeevani HB, Ujwala RK. In vitro protection of hepatocytes by Alocasia macrorrhiza leaf juice against CCl4 and tylenol mediated hepatic injury. International Journal of Pharmaceutical Applications. 2011;2(2):122-128




How to Cite

Nissar Ahmad Reshi, Sudarahana Mysore Shankarsingh, Girish Hodiyala Vasanaika. EVALUATION OF HEPATOPROTECTIVE POTENTIAL OF LEAF AND LEAF CALLUS EXTRACTS OF ANISOCHILUS CARNOSUS (L) WALL. ijp [Internet]. 2018 Sep. 30 [cited 2024 May 19];10(3):156-61. Available from: https://ijp.arjournals.org/index.php/ijp/article/view/603



Original Research Articles