Objective: Acalypha communis is a synonym of Ricinocarpus communis (MÃ¼ll.Arg.) belonging to the family of Euphorbiaceae. A. communis possess awide variety of activities like antimicrobial. Hence, the present study was intended to evaluate ethanolic leaf extract of A. communis for hepatoprotectiveactivities.Methods: The hepatoprotective activity was studied by paracetamol (2 g/kg) and D-galactosamine (400 mg/kg) induced models. Acute toxicity studyand preliminary phytochemical screening were also studied to evaluate the toxicity.Results: No toxicity profile was observed in rats after oral administration of the ethanolic leaf extract at the dose of 5 g/kg body weight. Thedifferent dose of 200 mg/kg and 400 mg/kg administered with the extract of A. communis. There was significant (p&lt;0.001) reduction in biochemicalparameters with respect to control. Phytochemical screening of the plant extract revealed the presence of tannins, alkaloids, flavonoids and saponins,and terpenoids.Conclusion: It can be concluded that the hepatoprotective activity elucidated by A. communis could be mainly due to the presences of high-value classof compound like the phenolic group as the major content in the plants.Keywords: Acalypha communis, Paracetamol, D-galactosamine, Biochemical parameters and histopathological studies

R, Anandan

S, RAJASEKARAN

English

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Vol 9, Issue 1, 2016 ISSN - 0974-2441PHYTOCHEMICAL AND PHARMACOLOGICAL EVALUATION OF ACALYPHA COMMUNIS MULL.ARG. FOR THEIR HEPATOPROTECTIVE ACTIVITYRAJASEKARAN S1*, ANANDAN R21Department of Pharmacology, Institute of Pharmaceutical Sciences and Research Center, Bhagwant University, Ajmer, Rajasthan, India. 2Department of Pharmacology, Erode College of Pharmacy, Erode, Tamil Nadu, India. Email: raj4ma@gmail.comReceived: 01 August 2015, Revised and Accepted: 29 August 2015ABSTRACTObjective: Acalypha communis is a synonym of Ricinocarpus communis (Müll.Arg.) belonging to the family of Euphorbiaceae. A. communis possess a wide variety of activities like antimicrobial. Hence, the present study was intended to evaluate ethanolic leaf extract of A. communis for hepatoprotective activities.Methods: The hepatoprotective activity was studied by paracetamol (2 g/kg) and D-galactosamine (400 mg/kg) induced models. Acute toxicity study and preliminary phytochemical screening were also studied to evaluate the toxicity.Results: No toxicity profile was observed in rats after oral administration of the ethanolic leaf extract at the dose of 5 g/kg body weight. The different dose of 200 mg/kg and 400 mg/kg administered with the extract of A. communis. There was significant (p<0.001) reduction in biochemical parameters with respect to control. Phytochemical screening of the plant extract revealed the presence of tannins, alkaloids, flavonoids and saponins, and terpenoids.Conclusion: It can be concluded that the hepatoprotective activity elucidated by A. communis could be mainly due to the presences of high-value class of compound like the phenolic group as the major content in the plants.Keywords: Acalypha communis, Paracetamol, D-galactosamine, Biochemical parameters and histopathological studies.INTRODUCTIONIndia has a rich culture of medicinal herbs and spices, which includes more than 2000 species and has a vast geographical area with high potential abilities for the Ayurvedic, Unani, and Siddha traditional medicines, but only very few have been studied chemically and pharmacologically for their potential medicinal value [1,2]. Hence, natural products from medicinal plants need to be investigated by scientific methods for their hepatoprotective activity. The plant Acalypha communis is a synonym of Ricinocarpus communis (Müll.Arg.) belonging to the family of Euphorbiaceae. It includes herbs, shrubs, and small trees, Shrubs or suffrutex frequently with resinous bright droplets on leaves and inflorescences; indumentum of simple or glandular hairs. Inflorescences spicate, usually unisexual [3].METHODSCollection, identification, and authentification of the plantsThe leaves of A. communis Müll.Arg., were collected from the Malappuram District, Kerala, India, during the month of October 2013. The plant materials were identified and authenticated by Dr. Pradeep Botanist Calicut University, Kozhikode. Voucher specimens were kept in our laboratory for future reference.Preparation of extractsThe granulated, dried leaves of A. communis (500 g) were packed in a Soxhlet apparatus and subjected to continuous hot percolation for 8 hrs using 450 ml of ethanol (95% v/v) as a solvent. The extract was concentrated to dryness under reduced pressure and controlled temperature and dried in a desiccator (yield 75 g, 15% w/w). The extract was suspended in 5% gum acacia and used for further experiments.Preliminary phytochemical screeningThe extract was screened qualitatively for the presence of various groups of phytoconstituents using different chemical tests [4].Procurement of experimental animalsAnimals were selected as per the OECD guidelines. Healthy young and nulliporous, non-pregnant Sprague Dawleys female rats weighing from 160 to 180 mg of 8-12 weeks old were selected, because literature survey of lethal dose 50% test shows that usually there is little difference in sensitivity between sexes, but generally females were found slightly more sensitive, were procured from listed suppliers of Sri Venkateswara Enterprises, Bengaluru, India. The animals were fed with standard pellet diet (Hindustan Lever Ltd. Bengaluru) and water ad libitum. All the animals were housed in polypropylene cages. The animals were kept under the alternate cycle of 12 hrs of darkness and light. The animals were acclimatized to the laboratory conditions for 1 week before starting the experiment. The animals fasted for at least 12 hrs before the onset of each activity. The experimental protocols were approved by Institutional Animal Ethics Committee (IAEC No.- P.Col/02/1868/26/09/2013/IAEC/JSPC) after scrutinization. The animals received the drug treatments by oral routs.OBSERVATIONSAnimals were observed individually for 48 hrs after dosing at the first 30 minutes, periodically and during the first 24 hrs, with special attention given during the first 4 hrs and daily thereafter, for a total of 14 days. Additional observations were also made if the animals continue to display signs of toxicity. Observations included were changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavior pattern. Observations were also made and checked for tremors, convulsions, salivation, diarrhea, lethargy, sleep, and coma. Results were tabulated in Table 1.Experimental procedure [5]The rats were divided into the five groups each containing six rats.Group I: Control rats, which fed normal diet and water.Group II: Rats treated with paracetamol (2 g/kg) for 28 days.Research ArticleAsian J Pharm Clin Res, Vol 9, Issue 1, 2016, 94-97 Rajasekaran and Anandan 95Group III:  Rats treated with silymarin (100 mg/kg) + paracetamol (2 g/kg) orally once daily for 28 days.Group IV:  Rats treated with AC (200 mg/kg, i.p.) + paracetamol (2 g/kg) once daily for 28 days.Group V:  Rats treated with AC (400 mg/kg, i.p.) + paracetamol (2 g/kg) once daily for 28 days.Experimental procedure [5]The rats were divided into the five groups each containing six rats.Group I: Control rats, which fed normal diet and water.Group II: Rats treated with D-galactosamine (400 mg/kg) for 28 days.Group III:  Rats treated with silymarin (100 mg/kg) + D-galactosamine (400 mg/kg) orally once daily for 28 days.Group IV:  Rats treated with AC (200 mg/kg, i.p.) + D-galactosamine (400 mg/kg) once daily for 28 days.Group V:  Rats treated with AC (400 mg/kg, i.p.) + D-galactosamine (400 mg/kg) once daily for 28 days.Statistical analysisValues were represented as mean ± standard error of mean of three parallel data’s.RESULTSPreliminary phytochemical screeningThe preliminary phytochemical analysis of fractions of A. communis shows the presence of steroids, alkaloids, flavonoids, glycosides, saponins, tannin, and carbohydrate.DISCUSSIONThe present study reveals the hepatoprotective activity of A. communis against paracetamol and D-galactosamine-induced hepatic damage in rats. Hepatotoxic drugs such as d-galactosamine and acetaminophen reduces liver functional capacity, which leads to an accumulation of waste products such as ammonia in the blood [5]. The results show that A. communis is effective in low and medium doses (200 mg/kg, p.o and 400 mg/kg, p.o) paracetamol is a commonly used as analgesic Table 1: Acute toxicity study of ethanolic extracts of leaves of A. communis based on OECD guidelines 423S. No Number of animals Dose in mg/kg Report1 3 5 No death2 3 50 No death3 3 300 No death4 3 2000 No death5 3 5000 No deathA. communis: Acalypha communisS. No. Group/drug Dose (mg/kg) SGOT (IU/L)SGPT (IU/L)ALP (IU/L)Total bilirubin (mg/dl)Total protein (mg/dl)n=6; values were expressed mean±SEM; Group II was compared to Group I. Groups III to V were compared to Group II. *p<0.01 versus D-galactosamine group: Significant; **p<0.001 versus D-galactosamine group: Highly significant data were analyzed by one-way ANOVA followed by Dunnett’s t-test. SEM: Standard error of mean. SGOT: Serum glutamic oxaloactetic transaminase, SGPT: Serum glutamic pyruvic transaminase, ALP: Alkaline phosphataseS. No. Group/drug Dose (mg/kg)SGOT (IU/L)SGPT (IU/L)ALP (IU/L)Total bilirubin (mg/dl)Total protein (mg/dl)n=6; values were expressed mean±SEM; Group II was compared to Group I. Groups III to V were compared to Group II. *p<0.01 versus paracetamol group: Significant; **p<0.001 versus paracetamol group: Highly significant data were analyzed by one-way ANOVA followed by Dunnett’s t-test. SEM: Standard error of mean. SGOT: Serum glutamic oxaloactetic transaminase, SGPT: Serum glutamic pyruvic transaminase, ALP: Alkaline phosphataseObservation EffectsGross activity Upto 3 hrs 3 ½ hrs 4 hrs 4 ½ hrs 5 hrs 5 ½ hrs 6 hrs 12 hrs 24 hrsRespiration + + + + + + + + +Writhing − − − − − − − − −Tremor − − − − − − − − −Convulsions − − − − − − − − −Hind limb paralysis − − − − − − − − −Sense of touch and sound + + + + + + + + +Salivation + + + + + + + + +Diarrhea − − − − − − − − −Mortality − − − − − − − − −Table 4: Results of the effects of biochemical markers of D‑galactosamine‑induced hepatic injury in rats1 Group I - normal control (NaCl 0.9% w/v) 5 ml/kg 64.23±1.31 77.52±1.04 19.17±1.35 1.39±1.42 6.98±0.252 Group II - D-galactosamine (400 mg/kg)* 167.70±1.07* 181.02±1.24* 42.83±2.01* 8.31±0.36* 3.66±1.46*3 Group III - silymarin+D-galactosamine 100 mg/kg+(400 mg/kg)** 67.74±1.93** 80.21±2.06** 18.64±1.27** 1.43±1.52** 7.09±0.68**4 Group IV - AC+D-galactosamine 200 mg/kg+(400 mg/kg)** 79.31±1.03** 91.21±1.62** 27.94±1.42** 2.13±1.41** 5.91±1.29**5 Group V – AC+D-galactosamine 400 mg/kg+(400 mg/kg)** 69.23±1.42** 83.71±2.47** 20.75±1.61** 1.83±0.12** 7.21±1.02**1 Group I - normal control (NaCl 0.9% w/v) 5 ml/kg 51.23±2.31 60.12±1.24 27.17±1.72 1.14±0.13 8.74±0.842 Group II - paracetamol (2 g/kg)* 193.60±1.74* 173.10±1.36* 58.75±1.32* 12.18±0.74* 4.89±1.31*3 Group III - silymarin+paracetamol 100 mg/kg+(2 g/kg)** 59.41±1.32** 56.78±2.32** 29.34±1.54** 1.27±0.22** 8.03±0.41**4 Group IV - AC+paracetamol 200 mg/kg+(2 g/kg)** 73.67±1.23** 67.31±1.14** 38.31±1.81** 2.68±0.35** 6.85±0.23**5 Group V - AC+paracetamol 400 mg/kg+(2 g/kg)** 61.23±2.13** 58.47±1.87** 30.14±1.22** 1.61±0.16** 7.91±0.13**Table 3: Results of the effects of biochemical markers of paracetamol induced hepatic injury in ratsA. communis: Acalypha communis, Table 2: Results of gross behavioral studies in rats on administration of A. communisAsian J Pharm Clin Res, Vol 9, Issue 1, 2016, 94-97 Rajasekaran and Anandan 96and antipyretic drug and is safe in therapeutic doses but produces fatal hepatic necrosis with toxic doses [6]. The toxic effect of paracetamol is due to oxidative damage induced by its metabolite, N-acetyl-p-benzoquinoneimine, produced by the action of cytochrome P-450 in the liver. This metabolite reacts with reduced glutathione (GSH) to yield non-toxic 3-GS-yl-PCM. Depletion of GSH causes the remaining quinone and other natural endogenous oxygen species to bind to cellular macromolecules leading to cell death [7]. D-galactosamine is a well-established hepatotoxicant, which is widely used model which closely resembles human viral hepatitis in its morphologic and functional characteristics therefore considered very useful for evaluation of hepatoprotection [8,9]. D-galactosamine hepatotoxicity is considered as an experimental model of acute hepatitis although it does not affect other organs [10]. D-galactosamine is known to selectively block the transcription and indirectly hepatic protein synthesis and as a consequence of endotoxin toxicity, it causes fulminant hepatitis. The toxicity of D-galactosamine results from inhibition of RNA and protein synthesis in the liver. The metabolism of D-galactosamine may deplete several uracil nucleotides including uridine diphosphate (UDP)-glucose, UDP-galactose and uridine triphosphate deficiency, which trapped in the formation of uridine-diphosho galactosamine. Accumulation of UDP-sugar nucleotide may contribute to the change in the rough endoplasmic reticulum and to the disturbance of protein metabolism. Intense galactosamine of the membrane structures is thought to be responsible for loss in the activity of ionic pumps. The impairment in the calcium pumps, with consequent increase in the intracellular calcium is considered to be responsible for cell death [11-14].This study shows that hepatic injury induced by paracetamol and D-galactosamine caused a significant rise in marker enzymes serum glutamic oxaloactetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP), and total bilirubin. The serum enzymes such as SGOT, SGPT, ALP, and total bilirubin of treated animals were significantly reduced (p<0.01) by 28 days pretreatment of ethanolic extract of leaves of A. communis at two dose levels 200 and 400 mg/kg p.o, when compared with paracetamol and D-galactosamine-treated control (Group II). From the result, it is clear that the drugs show dose-dependent activity. Histopathological observation also revealed that pretreatment with AC protected the animals from paracetamol and D-galactosamine-induced liver damage. The results indicate that the leaves of A. communis possess the hepatoprotective activity.CONCLUSIONFrom the present work, we conclude that species of A. communis are highly potential in biological activity. The preliminary screening of the samples revealed the presences of high-value class of compound like the phenolic group as the major content in the plants.dcbaedcbaeFig. 2: Histopathological studies of liver (D‑galactosamine‑induced), (a) Group I, (b) Group II, (c) Group III, (d) Group IV, (e) Group VFig. 1: Histopathological studies of liver (paracetamol induced), (a) Group I, (b) Group II, (c) Group III, (d) Group IV, (e) Group VAsian J Pharm Clin Res, Vol 9, Issue 1, 2016, 94-97 Rajasekaran and Anandan 97REFERENCES1. Sandhu DS, Heinrich M. The use of health foods, spices and other botanicals in the Sikh community in London. Phytother Res 2005;19(7):633-42.2. Gupta MP, Solís PN, Calderón AI, Guionneau-Sinclair F, Correa M, Galdames C, et al. 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PHYTOCHEMICAL AND PHARMACOLOGICAL EVALUATION OF ACALYPHA COMMUNIS MULL. ARG. FOR THEIR HEPATOPROTECTIVE ACTIVITY

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PHYTOCHEMICAL AND PHARMACOLOGICAL EVALUATION OF ACALYPHA COMMUNIS MULL. ARG. FOR THEIR HEPATOPROTECTIVE ACTIVITY

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