World Journal of Pharmaceutical Research SJIF Impact Factor 5.990

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1 Lakshman et al. SJIF Impact Factor Volume 4, Issue 9, Research Article ISSN HEPATOPROTECTIVE ACTIVITY OF AQUEOUS ETHANOLIC EXTRACT OF AERIAL PARTS OF BARLERIA GIBSONI ON PCT- INDUCED HEPATOTOXICITY IN RATS Lakshman Kumar D 1*, Sravani M 1, Dr.Venkatesh P 2, Dr. Hepcy Kala Rani D 2, Dr. Purushothaman M 3, Sucharitha P A M 1 1 Department of Pharmacognosy, PRRM College of Pharmacy, Kadapa, Andhra Pradesh, India. 2 Department of Pharmaceutical Chemistry, PRRM College of Pharmacy, Kadapa, Andhra Pradesh, India. 3 Department of Pharmaceutics, PRRM College of Pharmacy, Kadapa, Andhra Pradesh, India. Article Received on 10 July 2015, Revised on 03 Aug 2015, Accepted on 26 Aug 2015 *Correspondence for Author Mr. Lakshman Kumar D Department of Pharmacognosy, PRRM College of Pharmacy, Kadapa, Andhra Pradesh, India. ABSTRACT Barleria gibsoni belonging to the family Acanthaceae is commonly known as Neel Koranti. Several species of Barleria are reported for treatment of inflammation associated diseases like Gout, Rheumatism and swelling etc. So, the present work has been taken for phytochemical screening and to investigate Hepatoprotective activity. Natural remedies from medicinal plants are considered to be effective and safe alternative treatment for liver injury. The present study was conducted to evaluate the Hepatoprotective activity of aqueous ethanolic [30:70] extract of aerial parts of Barleria gibsoni Dalz.(AEEBG) in albino wistar rats. The study was conducted using the popular inducing agent Paracetamol (PCT) in Carboxy methyl cellulose (CMC) and silymarin was used as reference standard in the respective model to treat for 21 days. The effect was estimated by measuring the enzymatic levels. Phytochemical screening of AEEBG revealed the presence of phytocontituents like carbohydrates, phenolic compounds, tannins and flavonoids. The aqueous ethanolic extract of aerial parts of Barleria gibsoni Dalz. has shown very significant hepato protection against PCT- induced hepatotoxicity in albino wistar rats. Vol 4, Issue 09,

2 KEYWORDS: Barleria gibsoni, Hepato protective activity, Paracetamol, Carboxy methyl cellulose, silymarin. INTRODUCTION Herbs have been the "green medicines" of the Earth since time began. Their gentle ways are easy for us to absorb into our body, mind, and spirit. When we take herbs internally, they nourish us deep inside. Herbs function in gentle and holistic ways. Herbs can help us by: Providing many nutrients that are easily absorbed such as vitamins, minerals, and proteins. Liver is a key organ regulating homeostasis within the body by various functions. Liver injury caused by toxic chemicals and certain drugs has been recognized as a toxicological problem. Hepatotoxicity is one of very common ailment resulting into serious debilities ranging from severe metabolic disorders to even mortality. Plants derived natural products such as tannins, flavonoids, glycosides and carbohydrates have received considerable attention in recent years due to their diverse pharmacological properties including antioxidant and hepatoprotective activity. [1] PCT is a widely used antipyretic and analgesic which produces acute liver damage if over dose is consumed. [2] PCT is mainly metabolized in liver to excretable glucuronide and sulphate conjugates. However, the hepatotoxicity of PCT has been attributed to the formation of toxic metabolites when a part of PCT is activated by hepatic cytochrome P 450 to a highly reactive metabolite NAPQI [N-acetyl-p-benzoquinone imine]. NAPQI is initially detoxified by conjugation with GSH [Glutathione] to form mercapturic acid. However, when the rate of NAPQI formation exceeds the rate of detoxification by GSH, it oxidizes tissue macromolecules such as lipid or SH group of protein and alters the homeostasis of calcium after depleting GSH. Barleria gibsoni dalz.. known as neel-korant is a small unarmed shrub belongs to family Acanthaceae, was selected for the study. Leaves are glabrous green above and whitish underneath of great beauty. [3] Flowers are blue-pink with dark purple. Blotches at the base of upper two lobes of corolla. The aerial parts are quite effective as a remedy for inflammation. Roots infusion is used for the treatment of acidity. [4] MATERIALS AND METHODS Materials Plant collection The aerial parts of Barleria gibsoni was collected during flowering season in the month of March from S.V.University, Tirupathi. The plant was authenticated by Dr. Madhava chetty, Vol 4, Issue 09,

3 Toxonomist, S.V.University, Tirupathi, and voucher specimen was numbered and deposited in our Research lab of PRRM.College of pharmacy. Extraction The coarsely powdered aerial parts were extracted with aqueous ethanol by using Soxhlet apparatus until the phytoconstituents were completely exhausted. The aqueous ethanolic extract was evaporated through rotary evaporator under reduced pressure and labeled. Phytochemical investigation Phytochemical analysis of extract was carried out to find out the presence or absence of phytoconstituents viz tannins, glycosides, carbohydrates, flavonoids etc. [5] Experimental animals Wistar albino rats ( g) of either sex were obtained from Raghavendra enterprises, Chennai and they were housed under standard husbandry conditions, 23 ± 2 C temperature,12 h light/dark cycle with standard rat feed with water. Acute toxicity studies Acute oral toxicity studies were conducted to determine the LD 50 cut off value (mg/kg body weight) as per the OECD 2001 Guideline 423 and OPTT up and down procedure. [6] Assessment of hepatoprotective activity In the present investigation, rats (n=6) were randomized in to following groups and the pharmacological investigation was carried using PCT as inducing agent and the test AEEBG at dose levels of 200 and 400 mg/kg as hepatoprotective agent. In experiment the rats are divided into following groups with six animals in each group. Group I - Saline water per orally 21 days. Group II - PCT (0.1 ml/kg, p.o) + Vehicle (0.1 ml/kg, p.o) for 21 days. Group III - PCT (0.1 ml/kg, p.o) + silymarin (20 mg/kg, p.o) for 21 days. Group IV - PCT (0.1 ml/kg, p.o) + AEEBG (200mg/kg, p.o) for 21 days. Group V - PCT (0.1 ml/kg, p.o) + AEEBG (400mg/kg, p.o) for 21 days. Treatment with plant extract was started after 24 h of administration of inducing agent. [1] Biochemical analysis Blood was collected and serum was separated by allowing the blood samples to coagulate for 30 min at 37 C followed by centrifugation (300 rpm for 15 min) and subjected for Vol 4, Issue 09,

4 determination of biochemical parameters like serum bilirubin, SGPT, SGOT and ALP. Liver was dissected out, washed with ice cold phosphate Buffer saline (PBS) (0.1 M, P H 7.4) and 10% tissue homogenate used for different biochemical analysis. [7] STATISTICAL ANALYSIS The results are expressed as Mean ± SEM of four animals from each group. The data were evaluated by one-way ANOVA followed by TURKEY 'S multiple comparison tests. *P values <0.05 was considered statistically significant. RESULTS Preliminary Phytochemical screening: The various Phytoconstituents present in the extract was given in Table 1. AEEBG showed significant amounts of tannins, phenolic compounds, flavonoids and carbohydrates. Table 1: Qualitative Phytochemical determination of active constituents in Ethanolic extract of Barleria gibsoni(eebg). Acute toxicity studies Plant constituent Flavonoids Carbohydrates Tannins Glycosides Alkaloids Phenolic Compounds Saponins EEBG The EEBG did not exhibit any toxic effects up to 5000 mg/kg body weight on oral administration. Body weight before and after administration were noted and any changes in skin, eyes, mucous membrane, respiratory, circulatory, autonomic, central nervous system, behavioral pattern were observed, sign of tremors, convulsions, salivation, lethargy, sleep and coma were seen. The onset of toxicity and signs of toxicity were not seen in the rats up to 72 h of observation period. This indicates the extract is safe. Biochemical parameters Rats treated with PCT showed a significant hepatic damage as observed from elevated levels of hepato-specific enzymes as well as severe alteration in different liver parameters. SGPT, SGOT, and serum bilirubin and ALP in serum were increased in PCT intoxicated control animals. Treatment with the aqueous ethanolic extract of Barleria gibsoni showed significant Vol 4, Issue 09,

5 protection against both PCT induced liver damage against increase in serum enzyme levels and bilirubin in a dose responsive manner. Table-2: Aqueous Ethanolic extract of Barleria gibsoni (AEEBG) on serum enzymatic activity in PCT induced liver damage in rats (n=6). GROUPS SGOT(IU/L) SGPT(IU/L) ALP(IU/L) SB (mg/dl) I ± ± ± ± 1.18 II ± ± ± ± 2.04 III ± 11.23*** ± 1.43*** ± 4.6** 1.47 ± 2.41*** IV ± 1.9* ± 3.73* ± ± 3.70* V ± 10.8** ± 5.103** ± 7.2* 1.81 ± 1.16** Each value represents the mean±sem, n=6. * p<0.05, ** p<0.01, *** p<0.001, Statistical significant test for comparison was done by Turkey s multiple comparison tests. Groups III to V are compared with group II. SGOT: Serum Glutamate Oxaloacetate Transaminases, SGPT: Serum Glutamate Pyruvate Transaminases, ALP: Alkaline Phosphatases, SB: Serum bilirubin Fig 1: Effect of Barleria gibsoni on serum biochemical parameters values graph Fig 2: Effect of Barleria gibsoni on serum bilirubin Vol 4, Issue 09,

6 DISCUSSION PCT is a widely used antipyretic and analgesic which produces acute liver damage if overdoses are consumed. [8] PCT is mainly metabolized in liver to excretable glucuronide and sulphate conjugates. However, the hepatotoxicity of PCT has been attributed to the formation of toxic metabolites when a part of PCT is activated by hepatic cytochrome P-450 to a highly reactive metabolite NAPQI. NAPQI is initially detoxified by conjugation with GSH to form mercapturic acid. However, when the rate of NAPQI formation exceeds the rate of detoxification by GSH, it oxidizes tissue macromolecules such as lipid or SH group of protein and alters the homeostasis of calcium after depleting GSH Administration of EEBG effectively replenished the PCT induced depletion of hepatic GSH presumably due to diminished production of toxic metabolite, NAPQI through the inhibition of P-450 enzyme system. PCT induced damage of hepatocytes is also a reason behind decreased glycogen content of liver tissue. Significant increase in hepatic glycogen level was observed after administration of the extract indicating improvement in hepatic status. Oral administration of various doses of AEEBG to PCT intoxicated rats resulted in gradual normalization of the activities of SGOT, SGPT and ALP. This evidently suggests the protective effect of the extract in improving the functional integrity of liver cells. Serum bilirubin is considered as an index for the assessment of hepatic function and any abnormal increase indicates hepatobiliary disease and severe disturbance of hepatocellular architecture. PCT administration resulted in increased serum bilirubin level, thereby suggesting severe hepatic injury and confirming the hepatotoxic nature of PCT. Treatment with AEEBG significantly decreased the elevated level of total bilirubin in serum towards normally indicating its hepatoprotective efficacy. Hepatotoxins impair the capacity of liver to synthesize albumin. Decreased total serum protein level in PCT treated rats may be attributed to impaired protein synthesis by damaging liver tissue. Subsequent treatment of PCT intoxicated rats with AEEBG increased the total serum protein level. The curative efficacy of AEEBG was dose dependent as evidenced by gradual reversal of the altered values of various biochemical markers back to normal following oral administration. This may, probably be through promotional activation of anti oxidative enzymes and Vol 4, Issue 09,

7 regeneration of hepatocytes that restore the structural and functional integrity of liver. The protective effects due to treatment with Barleria gibsoni extract strongly indicated the possibility of the extract being able to prevent and/or mitigate any leakages of marker enzymes into circulation, condition the hepatocytes to accelerate regeneration of parenchymal cells, and preserve the integrity of the plasma membranes and hence restore these enzymes levels. Both the test groups i.e. low dose and high dose treated groups show dose dependent hepatoprotective activity. The test groups containing the plant extract alone showed an improvement in the liver Barleria gibsoni has the hepatoprotective activity. CONCLUSION As the literature survey reveals that works carried out for Barleria gibsoni is having many pharmacological activities, in which many of the activities are not yet proved. So this plant was taken for the thesis work. The aerial parts of the plant were taken and it was extracted using aqueous ethanol as solvent and preliminary phytochemical studies were performed in the extract and the chemical tests shows the presence of flavonoids, carbohydrates, tannins, phenolic compounds along with other components. The results obtained from the analysis of biochemical parameters studies conclude that the co-treatment of Barleria gibsoni extract prevents paracetamol induced hepatotoxicity in rats. The high dose Barleria gibsoni extract, showed better results as compared to low dose, both biochemically and morphologically, and the results were comparable with that of standard drug silymarin. Purifying the extract and identifying the active principles like flavonoids, tannins, carbohydrates, phenolic compounds may yield a good hepatoprotective drug. This study showed that AEEBG has a significant action against the hepatotoxicity induced by the drug used in the treatment of analgesic and antipyretic. The hepatoprotective role of AEEBG might be due to its antioxidant potential mechanism suggesting that the extract of plant may be useful to prevent the oxidative stress induced liver damage. On the basis of results obtained, it can be concluded that the aqueous ethanolic extract of Barleria gibsoni aerial parts were seems to produce hepatoprotective activity in rats. No toxic symptom or mortality was observed in 48 h of study in mice. These results seem to support the traditional use of this plant in protecting liver against hepatotoxicity. Thus the present study indicates that the aqueous ethanolic extract of aerial parts of Barleria gibsoni may be Vol 4, Issue 09,

8 used as an effective hepatoprotective agent. ACKNOWLEDGEMENT We are thankful to Management of P. Rami Reddy Memorial College of Pharmacy, Kadapa, A.P for providing the all facilities for carried out this work. REFERENCES 1. D.Sateesh Babu, J.P.Yanadaiah, D.Lakshman Kumar, K.Siva Shankar Prasad, S.Siva Shankar. Hepato protective activity of aqueous ethanolic extract of aerial parts of Givotia Moluccana on CCL 4 -induced Hepato toxicity in Rats, Journal of Global trends in Pharmaceutical sciences., 2014; 5(3): Nilesh Mehta, Michael R Pinsky. Drug-Induced liver injury e-medicine 2012; Available in: all. 3. Dr. K. Madhava Chetty and Rajanikanth, Flowering Plants of Chittor district, students offset printers, 2008; LakshmanKumar.D, SivaShankar.S, N.Vasudevareddy, Saisaran.S Anti Inflammatory activity on Barleria gibsoni in carragenan induced albino wistar rats, American Journal of Pharmatech research., 2014; 4(4). 5. Kokate CK. Practical Pharmacognosy, 4th Edition, Vallabhprakashan, Delhi., 1994; )4&doclanguage=en 7. Mallory HT, Evenlyn EA. The determination of bilirubin with photoelectric colorimeter. J Biol Chem., 1937; 119: Michalets EL. "Update: clinically significant cytochrome P-450 drug interactions". Pharmacotherapy., 1998; 18(1): PMID Vol 4, Issue 09,