Antioxidant and hepatoprotective activities of ethanolic extracts of leaves of Premna esculenta Roxb. against carbon tetrachloride-induced liver damage in rats

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INTRODUCTION

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threatening side effects.>@_{In contrast, traditional remedies}

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the treatment of liver ailments for a long period of time

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Antioxidant and Hepatoprotective Activities of Ethanolic

Extracts of Leaves of Premna esculenta Roxb. against Carbon

Tetrachloride-Induced Liver Damage in Rats

Mahmud ZA, Bachar SC

1

_{, Qais N}

Department of Clinical Pharmacy and Pharmacology, 1_{Department of Pharmaceutical Technology,}

Faculty of Pharmacy, University of Dhaka, Dhaka, Bangladesh

Address for correspondence: Dr. Nazmul Qais, E-mail: nqais@yahoo.com

Access this article online

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Website: www.jyoungpharm.in

DOI:

10.4103/0975-1483.104366 ABSTRACT

Premna esculenta5R[EIDPLO\9HUEHQDFHDHLVDVKUXEXVHGE\WKHHWKQLFSHRSOHRI&KLWWDJRQJ+LOO7UDFWV

RI %DQJODGHVK IRU WKH WUHDWPHQW RI KHSDWRFHOOXODU MDXQGLFH The present study was done to evaluate the hepatoprotective and the in vivo DQWLR[LGDQW DFWLYLW\ RI HWKDQROLF H[WUDFWV RI OHDYHV RI WKH SODQW LQ FDUERQ WHWUDFKORULGHLQGXFHGOLYHUGDPDJHLQUDWV+HSDWRWR[LFLW\ZDVLQGXFHGLQUDWVE\LSLQMHFWLRQRI&&O₄ diluted ZLWKROLYHRLOYYP/NJERG\ZHLJKWRQDOWHUQDWHGD\VIRUGD\V$IWHUGD\VRISUHWUHDWPHQWRIWHVW H[WUDFWV WKH ELRFKHPLFDO PDUNHUV VXFK DV 6HUXP *OXWDPDWH 2[DORDFHWDWH 7UDQVDPLQDVH 6*27 6HUXP *OXWDPDWH 3\UXYDWH 7UDQVDPLQDVH 6*37$ONDOLQH 3KRVSKDWDVH $/3 WRWDO SURWHLQ DQG DOEXPLQ ZHUH HVWLPDWHGIROORZHGE\WKHPHDVXUHPHQWRIOLYHUF\WRVROLFDQWLR[LGDQWHQ]\PHVVXFKDVVXSHUR[LGHGLVPXWDVH FDWDODVHDQGSHUR[LGDVH7KHGDWDZHUHDQDO\]HGXVLQJRQHZD\DQDO\VLVRIYDULDQFH$129$IROORZHGE\

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P. esculenta showed a potential hepatoprotective activity and the protective action might have manifested by UHVWRULQJWKHKHSDWLF62'FDWDODVHDQGSHUR[LGDVHOHYHOV7KHUHVXOWVMXVWLI\WKHWUDGLWLRQDOXVHRIWKLVSODQW in liver disorders.

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ureterolithiasis.>@_{,QWKHWUDGLWLRQDOV\VWHPRI UHPHGLHVWKH}

leaves of the plant are applied on the affected place for the treatment of arthritis and bacterial and fungal infections.

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leaves are one of the ingredients of a medicine used for

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chemical or pharmacological investigation except for the

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have reported earlier.>@_{7KHUHIRUHWRMXVWLI\WKHWUDGLWLRQDO}

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hepatoprotective activities.

MATERIALS AND METHODS

Drugs and chemicals

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Plant material

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Preparation of plant extracts

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Determination of the total phenolic content

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gallic acid.>@_{7RDQDOLTXRWRI WKHH[WUDFWP/VROXWLRQ}

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for 20 min at room temperature for color development.

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0.021, obtained from the gallic acid calibration curve prepared

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Determination of reducing power

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of the ethanolic extract of leaves prepared in methanol

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absorbance of the reaction mixture indicated a higher

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Experimental animals

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under standard environmental conditions (temperature

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care as found in the guidelines.>@_{7KHVWXG\SURWRFROZDV}

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Hepatoprotective activity

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received the ethanolic extract of P. esculentaOHDYHV((3(/ DWDGRVHRIDQGPJNJGD\UHVSHFWLYHO\7KHYHKLFOH RUWHVWGUXJVZHUHDGPLQLVWHUHGRUDOO\IRUVXFFHVVLYHGD\V

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catalase, and peroxidase.

Assessment of liver function

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estimating the biochemical parameters such as Serum

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based on the reference method of International Federation

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Antioxidant enzymes assays in liver

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Statistical analysis

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RESULTS

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Total phenolic contents

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Reducing capacity

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compared to the standard antioxidant, ascorbic acid.

Effect of EEPEL on SGOT, SGPT, SALP, total protein and albumin concentrations in CCl₄-induced liver damage in rats

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Absorbance

Concentration (μg/mL)

Series1

(5)

Effect of EEPEL on the levels of antioxidant enzymes in CCl₄-induced liver damage in rats

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DISCUSSION

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WKH OLYHU F\WRFKURPHP HQ]\PHV WR IRUP KLJKO\

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lipid peroxidation thus causing damage to cell membrane,

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lipid peroxidative degradation of the liver cell plasma

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blood stream, and estimating the activities of these serum

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group is a clear manifestation of antihepatotoxic effects of

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stimulating the regeneration process of damaged cells.

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Table 1: Total phenolics content of ethanolic extracts of Premna esculenta leaves

Sample Concentration

(μg/ml)

Absorbance (mean ± SD)

Gallic acid equivalent (mg/g of tested extract; mean ± SD)

EEPEL 250 0.276 ± 0.007 15.625 ± 0.486

EEPEL = Ethanolic extracts of Premna esculenta leaves

Figure 2:Reducing power of the ethanolic extract of leaves of Premna esculenta (EEPEL) and ascorbic acid (ASA) at different concentrations. The extract showed a moderate reducing capacity compared to that of the standard ascorbic acid

0 0.2 0.4 0.6 0.8 1 1.2 1.4

0 50 100 150 200 250 300

Absorbance

Concentration(μg/ml)

EEPEL

ASA

Table 2: Effect of the ethanolic extract of Premna esculentaOHDYHVRQVHUXPHQ]\PHVWRWDOSURWHLQVDQGDOEXPLQLQ &&O₄-induced liver damage in rats

Group Dose SGPT (IU/L) SGOT (IU/L) SALP (IU/L) TP (g/dL) ALB (g/dL)

Group I (vehicle control)

5 mL/kg (p.o.)

52.07±2.77 68.50±3.33 60.5±2.11 7.13±0.061 4.59±0.036

Group II (CCl₄ control)

1 mL/kg (i.p.)

104.69±2.67* 109.52±3.66* 138.08±3.69* 2.26±0.058* 1.22±0.027*

Group III (Silymarin)

100 mg/kg (p.o.)

47.98±2.54** (107.7) 58.16±3.33** (125.2) 62.64±1.86** (97.23) 6.91±0.213*** (95.48) 3.91±0.134*** (79.82)

Group IV (EEPEL)

200 mg/kg (p.o.)

65.56±2.76**(74.36) 59.79±3.54** (121.2) 56.50±3.05** (105.1) 4.85±0.141*** (53.18) 3.25±0.186*** (60.23)

Group V (EEPEL)

400 mg/kg (p.o.)

54.09±2.99**(96.16) 55.84±4.18** (130.86) 50.05±1.43** (113.50) 5.63±0.178*** (69.19) 3.51±0.099*** (67.95)

All values are expressed as means ± SEM (n = 5). Figures in parentheses are the percentage protections in individual biochemical parameters from their elevated values caused

by CCl4. The percent protection was calculated as follows: 100 × (Values of CCl4FRQWUROíYDOXHVRIVDPSOH9DOXHVRI&&O4FRQWUROíYDOXHVRIFRQWURO6*37 6HUXP

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normal control group (P < 0.001). **6LJQL¿FDQWUHGXFWLRQFRPSDUHGWRWKH&&O₄-treated controls (P < 0.001). ***6LJQL¿FDQWLQFUHDVHFRPSDUHGWRWKH&&O₄-treated controls (P <

(6)

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of the protective agents themselves.>@_{A previous}_in vitroDQWLR[LGDQWDFWLYLW\VWXG\RI HWKDQROLFH[WUDFWVRI

leaves of P. esculentaVKRZHGWKDWWKHH[WUDFWSRVVHVVHV

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scavenging activities.>@_{,QWKHSUHVHQWVWXG\WKHHWKDQROLF}

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preventing cell membrane oxidation.>@_{An increase}

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of P. esculenta DJDLQVW &&OLQGXFHG KHSDWRWR[LFLW\ LQ UDWV FRXOG EH DWWULEXWHG WR LWV DELOLW\ WR UHVWRUH WKH DQWLR[LGDQWHQ]\PHV62'FDWDODVHDQGSHUR[LGDVHVRI OLYHUF\WRVRORUWRWKHIUHHUDGLFDOVFDYHQJLQJDFWLYLW\RI WKHH[WUDFW7KHREVHUYHGKHSDWRSURWHFWLYHDFWLYLW\RI WKHOHDI H[WUDFWPD\EHGXHWRWKHSUHVHQFHRI SKHQROLF FRPSRXQGV>7DEOH@VXFKDVSKHQROVDQGWDQQLQVIRXQG LQ WKH SUHYLRXV SUHOLPLQDU\ SK\WRFKHPLFDO VFUHHQLQJ>@

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and antioxidant and hepatoprotective activities.>@

CONCLUSION

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that the ethanolic extracts of leaves of P. esculenta

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REFERENCES

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0LWUD6.6HVKDGUL6-9HQNDWDUDQJDQQD09*RSXPDGKDYDQ68GXSD98 6DUPD'1(IIHFWRI +'DKHUEDOIRUPXODWLRQLQJDODFWRVDPLQHLQGXFHG KHSDWRSDWK\LQUDWV,QGLDQ-3K\VLRO3KDUPDFRO

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and root extracts of Premnae sculenta 5R[E LQ 3ROR[DPHU LQGXFHG K\SHUOLSLGHPLFPLFHDQGUDWV2ULHQW3KDUP([S0HG :ROIH.:X;/LX5+$QWLR[LGDQWDFWLYLW\RI DSSOHSHHOV-$JULF)RRG

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Table 3: Effect of the ethanolic extract of Premna esculentaOHDYHVRQDQWLR[LGDQWHQ]\PHVLQ&&O₄-induced hepatotoxicity in rats

Group Dose SOD (units/mg of protein) Catalase (units/mg of protein) Peroxidase (units/mg of protein)

Group I (Vehicle control) 5 mL/kg (p.o.) 18.26 ± 0.447 121.1 ± 5.468 0.80 ± 0.043 Group II (CCl₄ control) 1 mL/kg (i.p.) 5.30 ± 0.294*** 28.58 ± 2.127*** 0.17 ± 0.014*** Group III (Silymarin) 100 mg/kg (p.o.) 17.36 ± 0.428** (93.05) 160.5 ± 2.75** (142.04) 0.77 ± 0.04** (95.23) Group IV (EEPEL) 200 mg/kg (p.o.) 9.96 ± 0.077** (35.95) 93.97 ± 6.67** (70.67) 0.34 ± 0.033* (26.98) Group V (EEPEL) 400 mg/kg (p.o.) 10.80 ± 0.335** (42.44) 111.06 ± 2.76** (89.15) 0.344 ± 0.037* (27.62)

Values are means ± SEM (n = 5), using one way ANOVA followed by Dunnett’s t-test. Figures in parentheses are the percentage restorations of antioxidant enzyme activity

from their reduced values caused by CCl4. The percent restoration was calculated as follows: 100 u (values of sample- values of CCl4 control)/ (values of control-values of CCl4

control). SOD = superoxide dismutase. *P < 0.01 when compared with the CCl₄ control group. **P < 0.001 when compared with the CCl₄ control group. ***P < 0.001 when

(7)

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