Full Length Research Article

Available online at http://www.ajol.info/index.php/njbas/index

Nigerian Journal of Basic and Applied Science (March, 2013), 21(1): 45-54 ISSN 0794-5698
DOI: http://dx.doi.org/10.4314/njbas.v21i1.7

Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against
Carbon Tetrachloride – Induced Hepatotoxicity in Rats

*1A. Nasir, 1,2M.G. Abubakar, 2R.A. Shehu, 3U. Aliyu, and 4B.K. Toge
1Department of Biochemistry, Umaru Musa Yar’adua University, Katsina, Nigeria.
2Department of Biochemistry, Usmanu Danfodiyo University Sokoto, Nigeria.
3Department of Crop Science, Usmanu Danfodiyo University, Sokoto, Nigeria.
4Department of Pharmaceutics, Delta State University, Abraka, Nigeria,

[*Corresponding author: Email [email protected]]

ABSTRACT: Hepatoprotective effect of the aqueous leaf extract of Andrographis paniculata was investigated
against CCl4 – induced hepatic injury in rats. Significant (P<0.05) increase of serum levels of alanine
aminotransferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), total bilirubin (TBL),
direct bilirubin (DBL), total cholesterol (CHL), triglycerides (TG), low density lipoprotein (LDL), very low density
lipoprotein (VLDL) and malondialdehyde (MDA) in CCl4 intoxicated rats were restored to normal levels when
treated with the extract and CCl4. Significant (P<0.05) decrease of serum levels of total protein (TP), albumin
(ALB), high density lipoprotein (HDL) and reduced glutathione (GSH) in CCl4 intoxicated rats were restored to
normal levels when treated with the extract and CCl4. The LD50 of the leaf extract was greater than 3000 mg/kg.
The study demonstrated that A. paniculata possesses significant hepatoprotective effects and may be the source
of lead compound in the management of liver diseases.
Key words: Hepatoprotective, Andrographis paniculata, Carbon Tetrachloride, Liver Disease.

INTRODUCTION Recently, the use of medicinal plants to cure various

Liver is one of the largest organs in human body and forms of liver diseases and dysfunctions is becoming
the chief site for intense metabolism and excretion increasingly popular and has received wide acceptance
(Ahsan et al., 2009). It is involved with almost all the (Oyagbemi and Odetola, 2010). Moreover, a large
biochemical pathways to growth, fight against disease, number of medicinal plants have been found to offer
nutrient supply, energy provision and reproduction some hepatoprotection (Handa and Sharma, 1990;
(Ward and Daly, 1999). So it has a surprising role in the Trivedi and Rawal, 2000; Ghosh et al., 2007; Prakash
maintenance, performance and homeostasis of the et al., 2008; Khan et al., 2009; Ahsan et al., 2009;
body (Ahsan et al., 2009). Hepatic damage is Adeneye et al., 2009; Oyagbemi and Odetola, 2010).
associated with distortion of these metabolic functions
(Wolf, 1999). Liver damage is associated with cellular Andrographis paniculata Nees is a herbaceous plant,
necrosis, increase in tissue lipid peroxidation and commonly known as “King of Bitters”, in the family
depletion of reduced glutathione levels. In addition, Acanthaceae (Jarukamjorn and Nemoto, 2008). A.
serum levels of many biochemical markers like paniculata grows erect to a height of 30 – 110 cm with
transaminases, alkaline phosphatase, bilirubin, glabrous leaves and white flowers with rose-purple
triglycerides and cholesterol are elevated in liver spots on the petals (Sabu, 2006). It grows abundantly in
disease (Mascolo et al., 1988). Liver diseases pose a southeastern Asia – India, Pakistan and Indonesia but
serious challenge to international public health (Ahsan is cultivated extensively in China, Thailand (Sandberg,
et al., 2009). Unfortunately, conventional or synthetic 1994), the East and West Indies, and Mauritius (Gupta
drugs used in the treatment of liver diseases are et al., 1990). Because of its well – known medicinal
inadequate and sometimes can have serious side properties, it is cultivated quite easily. A. paniculata has
effects (Rao et al., 2005). Moreover, there are still no been used in Asia to treat variety of chronic and
specific treatments in modern medicine that give infectious diseases (Alpha Omega Labs, 2008). It has
protection to the liver against damage or help to been traditionally used as a remedy against diabetes
regenerate hepatic cells (Chatterjee, 2000; (Norhaida, 1994), hypertension (Ahmad and Asmawi,
Chattopadhyay, 2003). 1993), inflammation (Patarapanich et al., 2007) and
cobra bite (Selvanayagam et al., 1994). Extensive

45
 Nasir et al.: Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against............

research revealed that A. paniculata has a surprisingly dryness at 40°C, producing a fine and chocolate colour
broad range of pharmacological effects and some of solid residue. The dried residue was scrapped, weighed
them are extremely beneficial and include anticancer and the percentage yield was calculated. The dried
(Kumar et al., 2004), anti HIV (Chang and Yeung, residue was stored in a capped bottle.
1988), anti AIDS (Stephen and Comac, 2000), From the dried residue, a fresh solution was prepared
hepatoprotective (Handa and Sharma, 1990) and on each day of the experiment.
immunostimulatory (Puri et al., 1993). Other
pharmacological effects include; antimalarial (Misra et Experimental animals
al., 1992), antityphoid (Sabu, 2006), antiviral (Wiart et Wistar rats weighing between 180 to 200 g were
al., 2005), antifungal (Sabu, 2006), antibacterial (Mishra purchased from Animal House, Department of
et al., 2009), antidiarrhoeal (Gupta et al., 1993), Biological Sciences of Usmanu Danfodiyo University,
antipyretic (Vedavathy and Rao, 1991), anti- Sokoto. The animals were kept in metal cages in a well
inflammatory (Chang and But, 1986), antidiabetic ventilated room and allowed to acclimatize for 14 days.
(Zhang and Tan, 2000), antithrombotic (Yeung et al., They were fed standard diet (Grand Cereals and Oil
1987), cardiovascular (Tan and Zhang, 2004), Mills Limited, Jos, Nigeria) and were provided clean tap
antivenom (Chang and But, 1986), antifertility (Sakila water ad libitum. The experiment was performed
et al., 2009) and psychopharmacological activity according to ethical guidelines of OECD (2001).
(Mandal et al., 2001). A. paniculata may be a
promising treatment for the alleviation of subjective Determination of LD50
symptoms of respiratory tract infections (Coon and Aqueous leaf extract of A. paniculata (3000mg/kg body
Ernst, 2004). It may also be beneficial for those with weight) was administered orally to five rats (one after
chronic fatigue syndrome and fibromyalgia (Khan, the other at an interval of 48 hours) in a single dose
2007). using oro-gastric tube. The control group received
distilled water. Observations of toxic symptoms were
The aim of the present study was to determine the LD 50 made and recorded systematically, at one, two, four
and the hepatoprotective effect of the aqueous leaf and six hours after administration. The number of
extract of A. paniculata against carbon tetrachloride survivors was noted after 48 hours for each group of
intoxicated rats. animals. The toxicological effects were assessed on the
basis of mortality and expressed as LD50 and calculated
MATERIALS AND METHODS using the limit test dose, up and down procedure of
Collection of plant material Organization for Economic and Cultural Development
Fresh leaves of A. paniculata plant were obtained from (OECD, 2001).
University Research Farm (Kwalkwalawa), Usmanu
Danfodiyo University Sokoto. The plant was identified at Hepatoprotective activity test
Botany Unit, of the same Institution. A voucher Carbon tetrachloride induction of hepatotoxicity was
specimen was also deposited in the Herbarium of the done according to procedures of Rao et al. (2005) with
same Institution for reference. some modifications.

Preparation of plant material Experimental design

The leaves of A. paniculata were thoroughly rinsed in A total of 30 rats were used. The rats were randomly
tap water twice. The leaves were open- air –dried under divided, into six groups of five rats each, as follows:
shade, cut into small pieces and pulverised into coarse Group A (normal control untreated rats) received 1ml
powder (using pestle and mortar). The powder was daily dose of liquid paraffin (1ml/kg body weight, per os)
stored in an airtight bottle until required. for five days.

Preparation of aqueous extract Group B, C, D, E, and F were administered standard

Two hundred (200g) of the powdered sample was diet and tap water on day one.
mixed with 2000ml of distilled water, in a conical flask.
The mixture was stirred severally covered overnight at Group B (induction control) were administered 30%
room temperature, filtered using Whatman filter paper carbon tetrachloride in liquid paraffin (1ml/kg body
(15 cm). The filtrate was evaporated to complete weight, i.p.) for four days (from day two to five).

46
 Nigerian Journal of Basic and Applied Science (March, 2013), 21(1): 45-54

Group C received 30% CCl4 in liquid paraffin (1ml/kg Table 2 shows that groups of rats treated with CCl4
body weight, i.p.) and Silymarin, a known demonstrated significant (p<0.05) increase in the serum
antihepatotoxic drug (Sigma Chemicals Company levels of total cholesterol (CHL), triglycerides (TG), low
USA), at a dose of 100mg/kg, per os, for four days density lipoprotein (LDL), very low density lipoprotein
(from day two to five). (VLDL) and malondialdehyde (MDA) when compared to
normal control rats. However, serum levels of high
Groups D, E, and F (test groups) were treated with 30% density lipoprotein (HDL) and reduced glutathione
CCl4 in liquid paraffin (1ml/kg body weight, i.p.) and a (GSH) were significantly reduced in CCl4- treated rats.
daily dose of 100, 200 and 300 mg/kg body weight
(orally) of aqueous leaf extract of A. paniculata Acute toxicity test at 3000 mg/kg body weight of
respectively, for four days (from day two to five). aqueous leaf extract of A. paniculata produced no
mortality after 48 hours of observation. The median
Clinical chemistry lethal dosage (LD50) of the aqueous leaf extract was
On the sixth day, all the animals were sacrificed under therefore estimated to be greater than 3000 mg/kg body
chloroform anaesthesia and blood and liver samples weight. The extract did not produce any negative
were collected. The blood collected was allowed to clot behavioural changes such as restlessness, excitement,
for 30 minutes. Serum was separated by centrifuging at respiratory distress, convulsions or coma. However, a
3000 rmp for 5 minutes. The supernatant was collected reduction in weight of the rats was observed. The
using Pasteur pipette into the sample bottles. The reduction in weight might be due to reduced food and
serum was used for biochemical estimations. Section of water intake, which might be secondary to feeling of
the liver was perfused with cold 0.86% KCl, fullness and loss of appetite after administration
homogenised, and centrifuged to obtain post (Joseph et al., 1989). Despite the listed side effects, the
mitochondrial supernatant for estimation of liver high value of the LD50 showed that the aqueous leaf
reduced glutathione and malondialdehyde content. The extract of A. paniculata was practically non-toxic.
activities of serum transaminases (ALT and AST),
alkaline phosphatase, albumin, total protein, bilirubin Carbon tetrachloride is a simple molecule which, when
(total and direct), total cholesterol, triglycerides and administered to a variety of species, causes
high density lipoprotein were assayed by Randox centrilobular hepatic necrosis and fatty liver. Low doses
(assay kit) methods of Reitman and Frankel (1957), of carbon tetrachloride cause only fatty liver and
Sood (1999), Doumas et al. (1971), Gornall et al. destruction of hepatic cytochrome P-450. However,
(1949), Jendrassik and Grof (1938), Trinder (1969), chronic administration or exposure leads to liver
Tietz (1990), and Lopez-Virella et al. (1977) cirrhosis and in some instances liver cancer and kidney
respectively. Serum levels of low density lipoprotein damage (Timbrell, 1987). Simultaneous administration
and very low density lipoprotein were calculated by of the plant extract and CCl4 produced an effect that
using Friedewald formula (Friedewald et al., 1972). was almost similar to that produced by silymarin. The
Post mitochondrial supernatant was used for the assay ability of a hepatoprotective drug to reduce the injurious
of reduced glutathione and malondialdehyde content by effects or to preserve the normal hepatic physiological
the methods of Patterson and Lazarow (1955) and mechanisms which have been disturbed by a
Hartman (1983) respectively. hepatotoxin is the index of its protective value (Yadav
and Dixit, 2003). The hepatotoxic effects induced by
RESULTS AND DISCUSSION CCl4 arise from its metabolite •CCl3, a free radical that
Groups of rats treated with CCl4 exhibited significant alkylates cellular proteins and other macromolecules
(p<0.05) increase in the activity of alanine with a simultaneous attack on polyunsaturated fatty
aminotransferase (ALT), aspartate amino transferase acids, in the presence of oxygen, to produce lipid
(AST), total bilirubin (TBL), direct bilirubin (DBL) and peroxides, leading to liver damage (Bishayee et
alkaline phosphatase (ALP) when compared to normal al.,1995; Nan et al., 2002). As shown in Table 1,
control rats. However, serum total protein (TP) and induction of hepatic damage with CCl4 was marked by a
albumin (ALB) were significantly (p<0.05) reduced in significant (P<0.05) increase in serum levels of marker
CCl4- treated rats (Table 1). enzymes ( ALT, AST and ALP) and bilirubin (TBL and
DBL) in the induction control rats (Group B) when
compared to normal control untreated rats (Group A).

47
 Nasir et al.: Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against............

Treatment with 100, 200 and 300 mg/kg (orally) of A. serum triglyceride, total cholesterol, HDL, LDL and
paniculata aqueous leaf extract was marked by VLDL as measuring parameters of liver function since
significant (P<0.05) decrease in serum levels of marker they are synthesised de novo in the liver. Significant
enzymes (ALT, AST and ALP) and bilirubin (TBL and (P<0.05) increase in CHL, TG, LDL and VLDL was
DBL), in a dose – dependent manner, when compared observed in the CCl4-intoxicated group. Significant
to Group B values. Induction with CCl4 was marked by (P<0.05) decrease in HDL was observed in CCl4-
a significant (P<0.05) decrease in serum levels of total intoxicated group. The inhibition of protein synthesis
protein (TP) and albumin (ALB). Treatment with 100, and disturbance of phospholipids metabolism might be
200 and 300 mg/kg (orally) of A. paniculata aqueous responsible for the abnormal levels of lipoproteins in the
leaf extract was marked by significant (P<0.05) serum. Treatment with aqueous leaf extract of A.
increase in serum levels of TP and ALB, in dose – paniculata significantly reversed these changes. This is
dependent manner, when compared to Group B. As an indication that the extract preserved hepatic protein
shown by the results, CCl4 doses induced acute hepatic synthesis and phospholipids metabolism.
damage as evidenced by a marked elevation in the
serum levels of the liver enzymes, ALT, AST and ALP, Enzymes in dose – dependent manner; demonstrating
and a significant decrease in the circulatory levels of TP A. paniculata aqueous leaf extract has hepatoprotective
and ALB, which are in conformity with earlier reports of effect. CCl4 induction was also associated with
the deleterious biochemical effects of CCl4 on hepatic significant decrease in the serum levels of albumin and
injury (Fadhel and Amran, 2002; Rajesh and Latha, total protein. However, treatment with A. paniculata
2004; Nagano et al., 2007). It is well documented in aqueous leaf extract protected the liver from the
literature that CCl4 is metabolised by mixed – function deleterious effect of the toxin by ameliorating the
oxidase system in the endoplasmic reticulum of the liver decrease in the circulatory levels of albumin and total
to the highly reactive trichloromethyl radical, and this protein in dose – dependent manner. CCl4 induction
reactive metabolite leads to auto – oxidation of the fatty causes degeneration of hepatocytes and blockade of
acids present in the cytoplasmic membrane the bile ducts which result into significant increase in
phospholipids and causes both functional and the serum levels of total bilirubin and direct bilirubin
morphological distortion of the cell membrane (Saraswat et al., 1993). Treatment with A. paniculata
(Recknagel and Glende, 1973). The hepatocyte aqueous leaf extract normalised the elevated serum
membrane distortion is associated with membrane levels of total bilirubin and direct bilirubin. Thus,
leakage of the hepatocyte cytosolic contents which is reduction in the levels of ALT and AST towards the
manifested by significant elevation of the serum marker normal value is an indication of regeneration process.
enzymes of acute hepatocellular damage namely ALT Reduction in the levels of ALP, total bilirubin and direct
and AST, and ALP as a marker for hepatobiliary bilirubin suggests the stabilisation of the biliary function.
damage (Bhattacharyya et al., 2003). However, of An increase in the serum levels of total protein and
these marker enzymes, ALT is the most reliable. AST is albumin suggests the stabilisation of endoplasmic
known to be present in abundance in the cardiac reticulum, leading to protein synthesis. Reduced
muscle, skeletal muscle, kidneys and testes, and ALP glutathione (GSH) is a naturally occurring substance
is abundant in the growing bone. Thus, any disease that is abundant in many living creatures. GSH is an
state affecting any of these extrahepatic tissues intracellular reductant and plays major role in catalysis,
significantly elevates the serum levels of these metabolism and transport (Ghosh et al., 2007). GSH
enzymes (Friedman et al., 1996). Extract treatment functions as free radical scavenger and in the repair of
significantly attenuated the acute elevation of these radical caused biological damage (Moron et al., 1979).
Table 2 shows the dose – dependent preventive effect GSH, through its significant reducing power, contributes
of oral doses of A. paniculata aqueous leaf extract to the recycling of other antioxidants such as vitamins C
against the deleterious effect of CCl4 intoxication on the and E that have become oxidised. Reduced GSH is a
serum lipid profile. The liver is known to be involved in critical determinant of tissue susceptibility to oxidative
the syntheses of triglyceride and cholesterol which are damage and the depletion of hepatic GSH has been
synthesised from a substrate, acetyl CoA (produced shown to be associated with an enhanced toxicity to
through fatty acid oxidation) (West et al., 1966). The chemicals, including CCl4 (Kidd, 1997).
hepatoprotective effects of the oral doses of A.
paniculata aqueous leaf extract were determined using

48
 Table 1: Effect of aqueous leaf extract of Andrographis paniculata on serum liver function indices in CCl4 – induced hepatic injury in rats

GROUP Treatment Dose (mg/kg ) ALT (U/l) AST (U/l) TP (g/dl) ALB (g/dl) TBL (µmol/l) DBL (µmol/l) ALP (U/l)

A Paraffin - 9.34 ± 0.26 9.56 ± 0.44 7.74 ± 0.10 4.18 ± 0.17 14.29 ± 0.24 2.46 ± 0.28 108.19 ± 3.20

B - - 14.66 ± 0.29 α 14.06 ± 0.38 α 5.65 ± 0.09 α 3.14 ± 0.17 α 21.87 ± 0.20 α 6.05 ± 0.23 α 304.15 ± 3.74 α

C Silymarin 100 9.38 ± 0.22 β 9.94 ± 0.42 β 7.52 ± 0.08 β 4.15 ± 0.11 β 14.43 ± 0.13 β 2.51 ± 0.28 β 114.26 ± 3.96 β

D APALE 100 10.98 ± 0.24 αβγ 11.02 ± 0.45 β 7.23 ± 0.08 αβ 3.84 ± 0.14 β 15.21 ± 0.19 β 3.89 ± 0.14 αβγ 142.97 ± 4.04 αβγ

49
E APALE 200 10.53 ± 0.16 αβγ 10.70 ± 0.48 β 7.29 ± 0.13 αβ 3.94 ± 0.13 β 14.97 ± 0.31 β 3.54 ± 0.25 αβ 127.51 ± 3.54 αβ

F APALE 300 10.05 ± 0.33 β 10.26 ± 0.56 β 7.38 ± 0.10 β 3.98 ± 0.16 β 14.58 ± 0.21 β 2.56 ± 0.23 β 120.34 ± 3.66 β

Values are means ± standard error of mean, n = 6, comparisons were made between:
‘’1’’- Group A vs B, C, D, E and F; ‘’2’’- Group B vs C, D, E and F; ‘’3’’- Group C vs D, E and F, using Bonferroni multiple comparisons test.
APALE= Andrographis paniculata aqueous leaf extract.
ALT= Alanine transaminase, AST= Aspartate transaminase, TP= Total protein, ALB= Albumin, TBL= Total bilirubin, DBL= Direct bilirubin, ALP= Alkaline phosphatase.
α = represents statistical significance vs A: p<0.05
β = represents statistical significance vs B: p<0.05
γ = represents statistical significance vs C: p<0.05
Nigerian Journal of Basic and Applied Science (March, 2013), 21(1): 45-54
 Table 2: Effect of aqueous leaf extract of Andrographis paniculata on lipid profile, GSH and MDA in CCl4–induced hepatic injury in rats

GROUP Treatment Dose CHL (mg/dl) TG (mg/dl) HDL LDL (mg/dl) VLDL GSH (mg/dl) MDA
(mg/kg ) (mg/dl) (mg/dl) (nmol/ml)
A Paraffin - 120.89 ± 116.40 ± 46.09 ± 51.53 ± 23.28 ± 218.57 ± 3.22 ± 0.02
B - - 212.44 ± 207.21 ± 31.44 ± 138.97 ± 41.44 ± 122.38 ± 5.53 ± 0.02 α
2.29 α 3.27 α 1.20 α 2.50 α 0.65 α 3.58 α
C Silymarin 100 121.33 ± 119.28 ± 42.70 ± 54.78 ± 23.85 ± 215.71 ± 3.26 ± 0.02 β
3.49 β 2.98 β 0.65 β 3.66 β 0.60 β 2.57 β

50
D APALE 100 147.56 ± 140.54 ± 41.35 ± 78.09 ± 28.11 ± 255.24 ± 3.47 ± 0.02
2.69 αβγ 2.85 αβγ 0.81 αβ 2.48 αβγ 0.57 αβγ 2.95 αβγ αβγ
E APALE 200 144.44 ± 138.38 ± 41.81 ± 74.96 ± 27.68 ± 264.29 ± 3.42 ± 0.02
6.00 αβγ 3.34 αβγ 0.72 αβ 6.48 αβγ 0.67 αβγ 2.26 αβγ αβγ
F APALE 300 143.56 ± 137.30 ± 42.48 ± 73.62 ± 27.46 ± 304.28 ± 3.31 ± 0.02 αβ
3.95 αβγ 2.57 αβγ 0.85 β 3.06 αβγ 0.51 αβγ 4.66 αβα
Values are means ± standard error of mean, n = 6, comparisons were made between:
‘’1’’- Group A vs B, C, D, E and F; ‘’2’’- Group B vs C, D, E and F; ‘’3’’- Group C vs D, E and F, using Bonferroni multiple comparisons test.
APALE= Andrographis paniculata aqueous leaf extract.
CHL= Total cholesterol, TG= Triglycerides, HDL= High density lipoprotein, LDL= Low density lipoprotein, VLDL= Very low density lipoprotein, GSH= Reduced
glutathione, MDA= Malondialdehyde.
α = represents statistical significance vs A: p<0.05
β = represents statistical significance vs B: p<0.05
γ = represents statistical significance vs C: p<0.05
Nasir et al.: Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against............
 Nigerian Journal of Basic and Applied Science (March, 2013), 21(1): 45-54

A significant (p<0.05) decrease in hepatic tissue GSH characterisation of the active principles responsible for
level was observed in the CCl4 treated group while hepatoprotective properties are recommended.
there was significant (p<0.05) increase in GSH level in
the groups treated with the plant extract. Hence, REFERENCES
exogenous aqueous leaf extract of A. paniculata Achiliya, G.S., Wadodkar, S.O. and Dorle, A.K. (2004).
supplementation might provide the means of recovering Evaluation of hepatoprotective effect of Amakadi
reduced GSH levels to prevent hepatic injuries. The ghrita against carbon tetrachloride induced
increase in hepatic GSH level in rats treated with the hepatic damage in rats. Journal of
extract and CCl4 might be due to de novo synthesis or Ethnopharmacology, 90: 229-232.
GSH regeneration. Adeneye, A.A., Olagunju, J.A., Elias, S.O.,
Olatunbosun, D.O., Mustafa, A.O., Adeshile,
The level of lipid peroxides is a measure of membrane O.I., Ashaolu, A.O., Laoye, T.A., Bamigboye,
damage and alterations in structure and function of A.O. and Adeoye, A.O. (2009). Protective
cellular membranes. A significant (p<0.05) elevation of activities of the aqueous root extract of
lipid peroxides in the liver of rats treated with CCl4 was Harungana madagascariensis in acute and
observed. The increase in MDA levels in the liver repeated acetaminophen hepatotoxic rats.
suggests enhanced lipid peroxidation leading to tissue International Journal of Applied Research in
damage and failure of antioxidant defence mechanisms Natural Products, 3: 29-42.
to prevent the formation of excessive free radicals Ahmad, M. and Asmawi, M.Z. (1993). Some
(Achiliya et al., 2004). Treatment with aqueous leaf pharmacological effects of aqueous extract of
extract of A. paniculata significantly reversed these Andrographis paniculata Nees. The International
changes. Conference on the Use of Traditional
Medicine and Other Natural Products in Health-
It has been earlier reported that decreasing the Care. (Abstracts), pp. 122.
metabolic activation of carbon tetrachloride, the Ahsan, R., Islam, M., Bulbul, J. I., Musaddik, A. and
antioxidant activity, prevention of generation of reactive Haque, E. (2009). Hepatoprotective activity of
oxygen species and scavenging of generated free Methanol extract of some medicinal plants
radicals or by combination of these are important against carbon tetrachloride-induced
mechanisms in the protection against CCl4-induced hepatotoxicity in rats. European Journal of
hepatic lesion (Yutin et al., 1990; Bhattacharyya et al., Scientific Research, 37(2): 302-310.
2003). The biochemical and antioxidant findings Alpha Omega Labs. (2008). Andrographis In-depth
obtained suggest that the extract might be mediating its Review. Alpha Omega Laboratories, Guayaquil,
protective effects either by decreasing the metabolic Ecuador. Retrieved November 5, 2009 from
activation of carbon tetrachloride, or by acting as a http://www.altcancer.com/andcan.htm
chain – breaking antioxidant for scavenging free Bhattacharyya, D., Mukherjee, R., Pandit, S., Das, N.
radicals or by combination of these effects. Moreover, and Sur, N.K. (2003). Prevention of carbon
previous studies have reported that protective effects of tetrachloride induced hepatotoxicity in rats by
hepatoprotective medicinal plants are mediated by their Himoliv, a polyherbal formulation. Indian Journal
flavonoids or alkaloids components or by their of Pharmacology, 35: 183- 185.
combination via antioxidant and free radicals Bishayee, A., Sarkar, A. and Chartterjee, M. (1995).
scavenging activities (Lanhers et al., 1991; Adeneye et The hepatoprotective activity of carrot (Daucus
al., 2009). The presence of secondary metabolites carota L.) against carbon tetrachloride in rats.
might thus be accounting for the hepatoprotective effect Journal of Pharmacy and Pharmacology, 54:
of A. paniculata aqueous leaf extract and could be via 555-563.
antioxidant or free radicals scavenging activities. Chang, H.M. and But, P.P.H. (1986). Pharmacology
and applications of Chinese material medica.
The study demonstrated that A. paniculata possesses Vol. 1. Singapore, World Scientific, pp.1918-928.
significant hepatoprotective effects and may be source Chang, R.S. and Yeung, H.W. (1988). Inhibition of
of lead compound in the management of liver diseases. growth of human immunodeficiency virus in vitro
Further studies regarding the isolation and

51
 Nasir et al.: Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against............

by crude extracts of Chinese medicinal herbs. medicinal plants against Escherichia coli
Antiviral Research, 9:163-175. enterotoxin-induced secretion in rabbit and
Chatterjee, T.K. (2000). Medicinal plants with guinea pig ileal loop models. Pharmaceutical
hepatoprotective properties. In: Herbal options, Biology, 31: 198-204.
3ed. Calcuta Books and Allied Publishers Handa, S.S. and Sharma, A. (1990). Hepatoprotective
Limited, pp.135-137. activity of andrographolide from Andrographis
Chattopadhyay, R.R. (2003). Possible mechanism of paniculata against carbontetrachloride. Indian
hepatoprotective activity of Azadirachta indica Journal of Medical Research, 92: 276-283.
leaf extract. Part II. Journal of Hartman, P.E. (1983). Putative mutagens and
Ethnopharmacology, 89(2-3): 217- 219. carcinogens in foods, IV. Malondialdehyde
Coon, J.T. and Ernst, E. (2004). Andrographis (Malondialdehyde). Environmental Mutagenesis,
paniculata in the treatment of upper respiratory 5 (4): 603-607.
tract infections: systematic review of safety and Jarukamjorn, K. and Nemoto, N (2008).
efficacy. Planta Medica, 70: 293-298. Pharmacological aspects of Andrographis
Doumas, B.T., Watson, W. and Briggs, H.G. (1971). paniculata on health and its major diterpenoid
Albumin standards and the measurement of constituent andrographolide. Journal of Health
serum albumin with bromocresol green. Clinica Science 54 (4):370-381..
Chimica Acta, 31: 87-96. Jendrassik, L. and Grof, P. (1938). Simplified
Fadhel, Z.A. and Amran, S. (2002). Effects of black tea photometric methods for the determination of
extract on CCl4-induced lipid peroxidation in bilirubin. Biochem Zschr., 297: 81-89.
liver, kidneys and testes of rats. Phytotherapy Joseph, P.K., Rao, K.R. and Sundaresh, C.S. (1989).
Research, 16 (Suppl.1): 28-32. Toxic effects of garlic extract and garlic oil in
Friedewald, W. T., Levy, R. I. and Fredrickson, D. S. rats. Indian Journal of Experimental Biology, 27:
(1972). Estimation of concentration of low 977-979.
density lipoprotein cholesterol in plasma without Khan, F. (2007). Andrographis – An Immune Boosting
use of preparative centrifuge.Clinical Herb. Retrieved January 22, 2010, from
Chemistry, 187: 589. http://ezinearticles.com/?Andrographis---An-
Friedman, L.S., Martin, P. and Muooz, S.J. (1996). Immune-- Boosting- Herb$id=538001
Liver function tests and the objective evaluation Khan, F.Z., Asdaq, S.M.B. and Kumar, P.S.R. (2009).
of the patient with liver disease. In: Hepatology: Effects of few Indian medicinal herbs on carbon
A Textbook of Liver Disease. Vol. 1, 3ed. tetrachloride induced injury in animals.
Philadelphia: WB Saunders Co., pp. 791-833. International Journal of Pharm. Tech
Ghosh, T., Maity, T.K., Das M., Bose, A. and Dash, Research, 1 (3). 579-587.
D.K. (2007). In vitro antioxidant and Kidd, P.M. (1997). Glutathione systemic protestant
hepatoprotective activity of ethanolic extract of against oxidative and free radical damage.
Bacopa monnieri Linn., aerial parts. Iranian Alternative Medicine Review, 2: 155-176.
Journal of Pharmacology and Therapeutics, 6: Kumar, R.A., Sridevi, K., Kumar, N.V., Nanduri, S. and
7-85. Rajagopal, S. (2004). Anticancer and
Gornall, A.G., Bardawill C.S. and David M.M. (1949). mmunostimulatory compounds from
Determination of serum proteins by means of Andrographis paniculata. Journal of
biuret reaction. Journal of Biological Chemistry, Ethnopharmacology, 92:291-295.
177: 751. Lanhers, M.C., Joyeux, M., Soulimani, R., Fleurentin,
Gupta, S., Choudhury, M.A., Yadava, J.N.S., J., Sayag, M., Mortier, F. And Younos, C.
Srivastava, V. and Tandon J. S. (1990). (1991). Hepatoprotective and anti-inflammatory
Antidiarrhoeal activity of diterpenes of effects of a traditional medicinal plant of Chile,
Andrographis paniculata (Kal-Megh) against Pneumus boldus. Planta Medica, 57: 110-115.
Escherichia coli enterotoxin in vivo models. Lopez – Virella, M.F., Stone, P., Ellis, S. and Colwell,
International Journal of Crude Drug Research, J.A. (1977). Cholesterol determination in high
28: 273-283. density lipoproteins separated by three different
Gupta, S., Yadava, J.N.S. and Tandon, J.S. (1993). methods. Clinical Chemistry, 23: 882-884.
Antisecretory (antidiarrhoeal) activity of Indian

52
 Nigerian Journal of Basic and Applied Science (March, 2013), 21(1): 45-54

Mandal, S.C., Dhara, A.K. and Maiti, B.C. (2001). planted in various seasons and regions in
Studies on psychopharmacological activity of Thailand. Thai Journal of Pharmaceutical
Andrographis paniculata extract. Phytotheraphy Sciences, 31:91-99.
Research, 15: 253-256. Patterson, J.W. and Lazarow, A. (1955). Methods of
Mascolo, N., Sharma, R., Jain, S.C. and Capasso, F. Biochemical Analysis. Glick D. (ed). New York,
(1988). Ethnopharmacology of Calotropis Inter Science, 2: 259.
procera flowers. Journal of Ethnopharmacology, Prakash, T., Faladu, S.D., Sharma, U.R., Surendra, V.,
22: 211-221. Goli, D., Stamina, P. and Kotresha, D.
Mishra, U.S., Mishra, A., Kumari, R., Murthy, P.N. and (2008). Hepatoprotective activity of leaves of
Naik, B.S. (2009). Antibacterial activity of Rhododendron arboretum in CCl₄ induced
ethanol extract of Andrographis paniculata. hepatotoxicity in rats. Journal of medicinal
Indian Journal of Pharmaceutical Sciences, Plants Research 2 (11). 315-320.
71: 436-438. Puri, A., Saxena, R., Saxena, R.P. and Saxena, K.C.
Misra, P., Pal, N.L., Guru, P.Y., Katiyar, J.C., (1993). Immunostimulant agents from
Srivastava, V. and Tandon, J.S. (1992). Andrographis paniculata. Journal of Natural
Antimalarial activity of Andrographis paniculata Products, 56: 995-999.
(Kalmegh) against Plasmodium berghei NK 65 Rajesh, M.G. and Latha, M.S. (2004). Protective activity
in Mastomys natalensis. International Journal of of Glycyrrhiza glabra Linn. on carbon
Pharmacognosy, 30: 263-274. tetrachloride – induced peroxidative damage.
Moron, M.S., Depierre, J.W. and Manmerik, B. (1979). Indian Journal of Pharmacology, 36: 284-287.
Level of glutathione, glutathione reductase and Rao, G.M.M., Rao, C.V., Pushpangadan, P. and
glutathione-s-transferase activity in rat lung and Shirwaikar, A. (2005). Hepatoprotective
liver. Biochimica et Biophysica Acta, 582: 67-78. effects of rubiadin, a major constituent of
Nagano, K., Umeda, Y., Saito, M., Nishizawa, T., Rubia cordifolia Linn. Journal of
Ikawa, N., Arito, H., Yamamoto, S. and Ethnopharmacology, 103 (3). 483-490.
Fukushima, S. (2007). Thirteen–week inhalation Recknagel, R.O. and Glende, E.A. Jr. (1973). Carbon
toxicity of carbon tetrachloride in rats and tetrachloride hepatotoxicity: An example of lethal
mice. Journal of Occupational Health, 49: 249- cleavage. CRC. Critical Reviews in Toxicology,
259. 2: 263-297.
Nan, J.X., Park E.J., Kim, V.C., and Sohn, D.H. (2002). Reitman, S. and Frankel, A.S. (1957). A colorimetric
Scutellaria baicalensis inhibits liver method for determination of serum glutamic
fibrosis induced by bile duct ligation or carbon oxaloacetic and glutamic pyruvic transaminase.
tetrachloride in rats. Journal of Pharmacy and American Journal of Clinical Pathology, 28: 56-
Pharmacology, 54:555-563. 63.
Norhaida, A.H. (1994). The effect of different fractions Sabu, K.K. (2006). Intraspecific variation of
of Andrographis paniculata extract on blood Andrographis paniculata Nees. (Doctoral thesis,
glucose level in diabetic rats. (Bachelor of University of Kerala, 2006). Retrieved from
Science Project Report). Department of Biology, http://sabuthesis.thesiencenet.com
UPM Malaysia, Malaysia. Sakila, S., Begum, N., Kawsar, S., Begum, Z.A., and
OECD (2001). Acute Oral Toxicity – Up – and - Down – Zoha, M.S. (2009). Relationship of anti-
Procedure (UDP). OECD Guidelines for the fertility effects of Andrographis paniculata and
Testing of Chemicals. hormonal assay in female rats. Bangladesh
Oyagbemi, A.A. and Odetola, A.A. (2010). Journal of Medical Science 8 (1-2). 10-14.
Hepatoprotective effects of ethanolic extract of Sandberg, F. (1994). Andrographidis herba
Cnidoscolus aconitifolius on paracetamol – Chuanxinlian. A Review. Swedish Herbal
induced hepatic damage in rats. Pakistan Institute, Gothenburg, Sweden.
Journal of Biological Sciences, 13(4). 164-169. Saraswat, B., Visen, P.K., Patnaik, G.K., Dhawan, B.N.
Patarapanich, C., Laungcholatan, S., Mahaverawat, N., (1993). Anticholestic effect of picroliv, active
Chaichantipayuth, C. and Pummangura, S. hepatoprotective principle of Picrorhiza kurroa,
(2007). HPLC determination of active diterpene against carbon tetrachloride induced cholestatis.
lactones from Andrographis paniculata Nees

53
 Nasir et al.: Hepatoprotective Effect of the Aqueous Leaf Extract of Andrographis paniculata Nees Against............

Indian Journal of Experimental Biology, 31: 316- Hills, Andhra Pradesh, India. Journal of
371. Ethnopharmacology, 33: 193-196.
Selvanayagam, Z.E., Gnavavendhan, S.G., Ward, F.M. and Daly, M.J. (1999). Hepatic Disease. In:
Chandrasekkharan, P., Balakrishna, K. and Clinical Pharmacy and Therapeutics (Walker R.
Rao, B.R. (1994). A review on And C. Edwards Eds.). Churchill Livingstone,
pharmacological and clinical studies. Fitoterapia New York, pp. 3346-3347.
65 (2). 99-111. West, E.E., Todd, W.R., Mason, H.S. and Van Bruggeu,
Sood, R. (1999). Medical Laboratory Technology, J.T. (1966). Textbook of Biochemistry, 4ed.
Methods and Interpretation. 5ed. Jaypee London: The Macmillan Company, pp. 1017-
Brothers Medical Publishers Ltd., pp. 488-490. 1118.
Stephen, H. and Comac, L. (2000). Miracle herbs: How Wiart, C., Kumar, K., Yusof, M.Y., Hamimah, H., Fauzi,
herbs combine with modern medicine to treat Z.M. and Suleman, M. (2005). Antiviral
cancer, heart disease, AIDS, and more, properties of ent-labdene diterpenes of
Kensington Publishing Corporation, New Andrographis paniculata Nees, inhibitors of
York. herpes simplex virus type 1. Phytotherapy
Tan, B.K.H. and Zhang, C.Y. (2004). Andrographis Research, 19: 1069-1070.
paniculata and the cardiovascular system. In Wolf, P.L. (1999). Biochemical diagnosis of liver
Herbal and Traditional Medicine: Molecular diseases. Indian Journal of Clinical Biochemistry
Aspects on Health. CRC Press, Teipei, pp. 441- 14, pp. 59-90.
456. Yadav, N.P. and Dixit, V.K. (2003). Hepatoprotective
Tietz, N.W. (1990). Clinical Guide to Laboratory Tests, activity of leaves of Khalanchoe pinnata pers.
2ed. W. B. Saunders Company, Philadelphia, Journal of Ethnopharmacology, 86: 197-202.
USA, pp. 554-556. Yeung, S.C.S., Yao, S.C. and Wang, L.L. (1987).
Timbrell, J.A. (1987). Principles of Biochemical Pharmacology and Applications of Chinese
Toxicology. Taylor and Francis Ltd, London, pp. Materia Medica, Vol. 2, World Scientific
140-143, 184-189. Publishing Co. Pte. Ltd., Singapore.
Trinder, P. (1969). A colorimetric method of Yutin, C., Rongliang, Z., Zhongjian, J. and Yong, J.
determination of serum and plasma (1990). Flavonoids as superoxide scavengers
cholesterol. Annals of Clinical Biochemistry, and antioxidants. Free Radicals Biology and
6: 24-27. Medicine, 9: 19-23.
Trivedi, N. and Rawal, U.M. (2000). Hepatoprotective Zhang, X.F. and Tan, B.K. (2000). Anti-diabetic
and toxicological evaluation of Andrographis property of ethanolic extract of Andrographis
paniculata on severe liver damage. Indian paniculata in streptozotocin-diabetic rats. Acta.
Journal of Pharmacology, 32: 288-293. Pharmacologica Sinica, 21:1157-1164.
Vedavathy, S. and Rao, K.N. (1991). Antipyretic activity
of six indigenous medicinal plants of Tirumala

54