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Research Article
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Normalisation of Lipoprotein Phenotypes by Chromolaena odorata-Linn. in Carbon Tetrachloride Hepatotoxicity-Induced Dyslipidaemia |
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C.S. Alisi,
A.O. Ojiako,
G.O.C. Onyeze
and
G.C. Osuagwu
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ABSTRACT
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The major functions of the liver can be detrimentally altered by liver injury resulting from acute or chronic exposure to toxicants. Dyslipidemia is often found in such toxicity resulting from chemical damage. Normalisation of atherogenic indices by Chromolaena odorata (C. odorata ) in carbon tetrachloride-induced liver toxicity was evaluated in 30 male rabbits divided into 5 groups of 6 animals each. Normal Control (NC) received food and water only. Carbon tetrachloride intoxicated control (CCl4) received a single dose of CCl4 (0.2 mL kgbw-1 in liquid paraffin 1:1). C. odorata test animals (ETECO TEST) received a single dose of CCl4 + ethanol extract of C. odorata at 400 mg/kg/day in two divided doses of 200 mg kg-1 morning and night, for 6 days. C. odorata control animals (ETECO CTRL) received ethanol extract of C. odorata at 400 mg/kg/day in two divided doses of 200 mg kg-1. Group five (Sylimarin) received sylimarin 50 mg/kgbw prior to CCl4 intoxication. Carbon tetrachloride-induced toxicity resulted in liver injury which was seen from the significant (p<0.05) elevation of the activities of serum Aspartate Aminotransferase (AST), Alanine aminotransferase (ALT), Lactate Dehydrogenase (LDH) and gamma-Glutamyl Transferase (γ-GT), significantly decreased protein and albumin and significantly increased total bilirubin concentrations; altered lipid and lipoprotein phenotypes in favour of increased atherogenic indices. Pre-treatment with C. odorata extract prevented these biochemical alterations and normalized the lipoprotein phenotypes. C. odorata may be useful not only as a hepatoprotective agent, but also in the reduction and/or prevention of adverse cardiovascular events.
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Received: June 06, 2011;
Accepted: June 28, 2011;
Published: December 02, 2011
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INTRODUCTION
The liver is a versatile organ in the body having a specialized function, reflected
in its anatomy and metabolic activity. It regulates internal chemical environment
(Gole et al., 1997) and plays a central processing
and distributive role in metabolism. The liver plays a significant role in the
body as the organ saddled with the responsibility of metabolising toxic substances
that enter the body. The major functions of the liver can be detrimentally altered
by liver injury (Singh et al., 2011) resulting
from acute or chronic exposure to toxicants (Khadr et
al., 2007) or by situations affecting both β-oxidation and the
respiratory chain enzymes. Altered free fatty acid metabolism and impaired aerobic
respiration as found in animals on high fat diet, have frequently been associated
with accumulation of lactate and Reactive Oxygen Species (ROS). The presence
of ROS further disrupts mitochondrial DNA and brings about the damage to hepatic
cells. Still other pathways to injury may develop when drugs damage mitochondria,
disrupting fatty-acid oxidation and energy production. When drugs bind to and
disable respiratory-chain enzymes, increased formation of ROS in mitochondria
results. This increases oxidative stress and a secondary attack on mitochondrial
DNA, with ensuing anaerobic metabolism, lactic acidosis and triglyceride accumulation
(microvesicular fat within cells) (Pessayre et al.,
2001).
Carbon tetrachloride (CCl4) has been one of the most intensively
studied hepatotoxicants to date and provides a relevant model for other halogenated
hydrocarbons that are used widely (Dahm and Jones, 1996;
Weber et al., 2003; Adinarayana
et al., 2011). It consistently produces liver injury in many species,
including non-human primates and man (Achudume and Ogunyemi,
2007; Moundipa et al., 2007; Noori
et al., 2009). Carbon tetrachloride is well known to be converted
by cytochrome P-450-mixed function oxygenases in smooth endoplasmic
reticulum of liver into toxic metabolite, mainly trichloromethyl radical (CCI3●).
This free radical in the presence of oxygen may cause peroxidation of lipids
on target cell resulting in extensive damage. Liver damage (hepatotoxicity)
remains one of the most serious health problems (Lee, 2003).
Antioxidation agents of natural origin have attracted special interest because
they can protect human body from free radical damage (Raja
et al., 2007; Babu et al., 2001; Gupta
et al., 2006) and can also protect the liver from damage initiated
by hyperlipidaemia (Alisi et al., 2008).
Numerous medicinal plants and their formulations are used for liver disorders
in ethno-medical practices (Chavda et al., 2010)
as well as in traditional systems of medicine in Africa, India and indeed Asia
(Yen et al., 2001; Rao et
al., 2006; Koneru et al., 2011). Chromolaena
odorata (L.) R. KING and H. ROBINSON (formerly Eupatorium odoratum L.),
a perennial belonging to the plant family Asteraceae (Compositae), is a diffuse,
scrambling shrub that is mainly a weed of plantation crops and pastures of Southern
Asia and Western Africa. This common plant is called Siam weed. The plant is
known among the Igbos of the South-Eastern Nigeria as Elizabeth Independence
leaf, Enugu plantation weed and Awolowo leaf (Alisi
and Onyeze, 2009). Results of a number of studies showed that the extract
of the leaves of C. odorata inhibited the growth of some bacteria (Irobi,
1997). Enhancement of haemostasis and blood coagulation with use of C.
odorata extract has also been reported. C. odorata has demonstrated
anti-inflammatory, astringent and diuretic activities (Owoyele
et al., 2006; Rao et al., 2010). We
already demonstrated a nitric oxide scavenging ability of ethyl acetate fraction
of methanol leaf extracts of Chromolaena odorata (Alisi
and Onyeze, 2008). Biochemical mechanisms of wound healing using ethanol
extract of Chromolaena odorata have also been previously reported. The
ethyl acetate fraction of methanol extract of C. odorata has been demonstrated
in vitro to scavenge hydroxyl radicals (Alisi and
Onyeze, 2009).
Patients with hyperlipidaemia have elevations in aminotransferases levels due
to non-alcoholic fatty liver disease (Mayes and Botham, 2003).
It is well known that hyperlipemia induces liver damage (Mukai
et al., 2002; Milionis et al., 2004).
Dyslipidaemia accompanies Carbon tetrachloride-induced liver toxicity. Dyslipidemia,
as low HDL-cholesterol, high Triglyceride (TG) and elevated LDL-cholesterol,
increase cardiovascular disease risk (Wilson et al.,
1991; Austin et al., 1998). Low density lipoprotein
cholesterol (LDL), HDL and TG are all independent and significant predictors
of cardiovascular risk. High blood cholesterol is one of the greatest risk factors
contributing to the prevalence and severity of coronary heart disease (Wilson
et al., 1998). The Total Non HDL Cholesterol (TNH-CHOL) is the single
greatest predictor of cardiovascular risk and can be used as a surrogate measure
of lowering the cardiovascular risk. In addition, there is a well known association
between hypertension and dyslipidemia, particularly high levels of serum LDL
cholesterol. Furthermore, the magnitude of the reduction in cardiovascular events
is a function of the extent of LDL cholesterol lowering. A decrease of 40 mg
per deciliter (1.0 mmol L-1) in LDL cholesterol will correspond to
a 24% reduction in major cardiovascular events (Baigent
et al., 2005). The adoption of crude extracts of plants, such as
infusions, for self-medication by the general public (Houghton,
1995), has arisen in the possibility that the impact of several diseases
may be either ameliorated or prevented by improving the dietary intake of natural
nutrients (Raja et al., 2007). We are not aware
of any studies on the normalisation of atherogenic indices by Chromolaena
odorata in carbon tetrachloride-induced liver toxicity. The purpose of this
study was to evaluate hypolipidaemic vis-α-vis antihepatic effects of Chromolaena
odorata extracts in carbon tetrachloride-induced liver damage.
MATERIALS AND METHODS Collection and preparation of plant samples: Fresh aerial parts of Chromolaena odorata were collected from Egbu and Ihiagwa in Owerri, Imo State (2008) and authenticated by a plant taxonomist, at the Department of Plant Science and Biotechnology, Imo State University, Owerri, Nigeria. Voucher specimen has been retained at the authors laboratory. The leaves were shed, dried at 30°C and then reduced to a coarse powder in a mill (Kenwood BL357). A 500 g portion was extracted with 2 L ethanol by shaking for 48 h. Soluble extract was recovered by distillation under reduced pressure at 49°C in a Buchi rotavapour (Switzerland). The extract was then dried to solid form in vacuum desiccator (CNS Simax) and stored in a freezer (4.0°C) until used. Animals: Thirty white New Zealand male rabbits acquired from an animal breeder in Owerri, Imo State, Nigeria were maintained under standard environmental condition (28-30°C, 60-70% relative humidity, 12 h dark/light cycle) in stainless steel cages with free access to standard laboratory animal diet (Vital finisher) and drinking water. Induction of hepatic injury: Seven days after acclimatization, animals were distributed into five groups of six animals each. Group I served as Normal Control (NC) which received food and water only throughout the treatment period. Group II served as intoxicated controls (CCl4 group) which received food and water ad libitum and carbon tetrachloride (0.2 mL kg b.wt.-1 in liquid paraffin 1:1) on day 7. Groups III served as intoxicated tests (ETECO test) that received food and water ad libitum, received ethanol extracts of C. odorata (400 mg kg-1 body weight of animal) in two divided equal daily doses and carbon tetrachloride (0.2 mL kg b.wt.-1 in liquid paraffin 1:1) on day 7. Group IV received food and water ad libitum and received ethanol extracts of C. odorata (400 mg kg-1 body weight of animal) in two divided equal daily doses but did not receive carbon tetrachloride. Group V received food and water ad libitum and received Sylimarin (50 mg kg-1 body weight of animal) daily and CCl4 on day 7. At the end of the seven-day pre-treatment and subsequent intoxication with carbon tetrachloride, animals were allowed for 48 h. Animals were anaesthetized and sacrificed by cervical dislocation as permitted by University Ethical Committee.
Biological assays: Blood collected from the ear blood vessels under
mild chloroform anesthesia was kept for 45 min at 4°C to clot. Serum was
separated by centrifugation at 600 g for 15 min and analyzed for various biochemical
parameters. Serum enzyme activities (aspartate aminotransferase (AST), Alanine
aminotransferase (ALT), gamma-Glutamyl Transferase (γ-GT), Lactate Dehydrogenase
(LDH), were assayed using a chemistry analyzer (Ciba-Corning 550 Express Plus.
USA). Total bilirubin estimation exploits the use of deoxidized sulphanilic
acid as described by Pearlman and Lee (1974). The Biuret
method as described by Gornall et al. (1949)
was employed for the determination of protein concentration in serum. Serum
albumin concentration was estimated by the method employing bromocresol green
as described by Doumas et al. (1971).
Total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol concentrations were measured spectrophotometrically (Pharmacia LKB Ultospec III) using assay kits (Biosystems S.A. costa Brava Barcelona Spain) while LDL/HDL-ratio and the total non-HDL cholesterol concentration (TNHCHOL) were simply calculated. Statistical analysis: Results of groups are calculated as Means±SD. and subjected to one-way Analysis of Variance (ANOVA). Significant difference between means were determined at alpha = 0.05. Analysis was done using Analyst v.2.2 (Leeds UK) on Microsoft Excel platform. RESULTS Effect of ethanol extract of C. odorata on plasma alanine amino transferase (ALT) Activity in carbon tetrachloride induced hepatotoxicity: Result showed that ALT activity in Carbon tetrachloride-intoxicated animals were elevated significantly (p<0.05) (115.93±11.8 U L-1) when compared to normal control (40.86±4.0 U L-1), ethanol extract of C. odorata control (62.72±7.2 U L-1) and ethanol extract of C. odorata treated group (77.88±6.8 U L-1). Sylimarin (52.6±4.4 U L-1) was however, more effective in protecting the hepatocytes against Carbon tetrachloride-induced damage (Fig. 1a). Effect of ethanol extract of C. odorata on plasma aspartate amino transferase (AST) activity in carbon tetrachloride-induced hepatotoxicity: Result showed that AST activity in Carbon tetrachloride-intoxicated animals was elevated significantly (p<0.05) (194.0±17.8 U L-1) when compared to normal control (31.0±3.2 U L-1), ethanol extract of C. odorata control (47.0±3.9 U L-1) and ethanol extract of C. odorata treated group (143.±15.4 U L-1). Sylimarin (80.82±10.5 U I-1) was however, more effective in protecting the hepatocytes against Carbon tetrachloride-induced damage (Fig. 1b). Effect of ethanol extract of C. odorata on lactate dehydrogenase (LDH) activity in carbon tetrachloride-induced hepatotoxicity: Result showed that LDH activity in carbon tetrachloride intoxicated animals was elevated significantly (p<0.05) (4833±540 IU L-1) when compared to normal control (2740±294 IU L-1), ethanol extract of C. odorata control (2992±246 IU L-1) and ethanol extract of C. odorata treated group (3543±600 IU L-1). Ethanol extract of C. odorata was comparable to sylimarin (3200±85 IU L-1) in restoring the LDH activity to normalcy (Fig. 1c).
Effect of ethanol extract of C. odorata on gamma glutamyl transferase
(γ-GT) activity in carbon tetrachloride-induced hepatotoxicity: Result
showed that γ-GT activity in carbon tetrachloride intoxicated animals was
elevated significantly (p<0.05) (26.0±4.5 IU L-1) when
compared to normal control (8.33±1.12 IU L-1), ethanol extract
of C. odorata control (11.67±2.5 IU L-1) and ethanol
extract of C. odorata treated group (13.0±2.16 IU L-1).
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Fig. 1 (a-d): |
Effect of Ethanol extract of C. odorata (400 mg kg-1)
on some serum indicators (enzymes) of liver injury (a) ALT, (b) AST, (c)
LDH and (d) γ-GT |
Table 1: |
Effect of ethanol extract of C. odorata (400 mg kg
bw-1) on some liver function parameters |
 |
Values are Mean±S.D. (n = 6). *p<0.05 vs. normal
control. p<0.05 vs. CCl4 control |
Sylimarin (10.3±2.9 IU L-1) was comparable to ethanol extract
of C. odorata in restoring the γ-GT activity to normalcy (Fig.
1d).
Effect of ethanol extract of C. odorata on total bilirubin concentration in carbon tetrachloride-induced hepatotoxicity: Result showed that total bilirubin concentration in carbon tetrachloride intoxicated animals was elevated significantly (p<0.05) (40.34 μmol L-1) when compared to normal control (15.34 μmol L-1), ethanol extract of C. odorata control (14.77 μmol L-1) and ethanol extract of C. odorata treated group (19.31 μmol L-1). Ethanol extract of C. odorata was comparable to sylimarin (16.43±1.70 μmol L-1) in restoring the total bilirubin concentration to normalcy (Table 1). Effect of ethanol extract of C. odorata on serum protein concentration in carbon tetrachloride induced hepatotoxicity: Table 1 showed that total protein concentration in carbon tetrachloride intoxicated animals was significantly (p<0.05) decreased (50.20±2.5 gl L-1) when compared to normal control (76.6±3.83 g L-1), ethanol extract of C. odorata control (68.81±3.5 g L-1) and ethanol extract of C. odorata treated group (57.56±3.4 g L-1).
Effect of ethanol extract of C. odorata on serum albumin concentration
in carbon tetrachloride induced hepatotoxicity: Table 1
showed that total albumin concentration in carbon tetrachloride intoxicated
animals was significantly (p<0.05) decreased (32.63±1.87 gl L-1)
when compared to normal control (49.76±1.47 g L-1), ethanol
extract of C. odorata control (44.73±3.32 g L-1) and
ethanol extract of C. odorata treated group (43.92±2.86 g L-1).
Effect of ethanol extract of C. odorata on serum total cholesterol concentration of carbon tetrachloride-induced hepatotoxicity: Figure 2a showed that total cholesterol concentration in carbon tetrachloride intoxicated animals was significantly (p<0.05) increased (6.89±1.31 mmol L-1) when compared to normal control (3.22±0.19 mmol L-1) and ethanol extract of C. odorata control (3.07±0.26 mmol L-1). Ethanol extract of C. odorata treated group (4.95±0.37 mmol L-1) had significantly lower cholesterol concentration than the intoxicated group, but were significantly (p<0.05) higher than normal controls. Effect of Ethanol extract of C. odorata on serum triacylglycerol concentration of carbon tetrachloride-induced hepatotoxicity: Figure 2b showed that serum triacylglycerol concentration in carbon tetrachloride intoxicated animals was significantly (p<0.05) increased (1.56±0.22 mmol L-1) when compared to normal control (1.015±0.056 mmol L-1) and ethanol extract of C. odorata control (0.766±0.07 mmol L-1). Ethanol extract of C. odorata treated group (1.094±0.1 mmol L-1) had significantly lower triacylglycerol concentration than the intoxicated group, but were significantly (p<0.05) higher than ethanol extract of C. odorata controls. Effect of ethanol extract of C. odorata on serum LDL-cholesterol concentration of carbon tetrachloride-induced hepatotoxicity: Figure 2c showed that serum LDL-cholesterol concentration in carbon tetrachloride intoxicated animals was significantly (p<0.05) increased (2.17±0.13 mmol L-1) when compared to normal control (1.37±0.11 mmol L-1) and ethanol extract of C. odorata control (1.726±0.17 mmol L-1). ethanol extract of C. odorata treated group (1.698±0.212 mmol L-1) had significantly lower LDL-cholesterol concentration than the carbon tetrachloride intoxicated group but were significantly (p<0.05) higher than normal controls. Effect of ethanol extract of C. odorata on serum HDL- cholesterol concentration of carbon tetrachloride-induced hepatotoxicity: Figure 2d showed that serum HDL-cholesterol concentration in carbon tetrachloride-intoxicated animals was significantly (p<0.05) decreased (1.003±0.008 mmol L-1) when compared to normal control (2.47±0.002 mmol L-1) and ethanol extract of C. odorata control (2.16±0.005 mmol L-1). Ethanol extract of C. odorata treated group (1.697±0.003 mmol L-1) had significantly (p<0.05) higher HDL-cholesterol concentration than the intoxicated group but were significantly lower than normal controls.
Effect of ethanol extracts of C. odorata on serum total non-HDL cholesterol
of carbon tetrachloride-induced liver damage: Intoxication of rabbits with
carbon tetrachloride produced an elevated total non-HDL-cholesterol (5.88 mMol
L-1) which was significantly (p<0.05) higher than the normal controls
(0.742 mMol L-1) and the ethanol extract of C. odorata treated
controls (0.910 mMol L-1) (Fig. 2e). Treatment
with ethanol extract of C. odorata significantly (p<0.05) lowered
TNHC (3.25 mMol L-1) in the intoxicated animals.
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Fig. 2 (a-f): |
Effect of ethanol extract of C. odorata on serum lipid/lipoprotein
phenotype of carbon tetrachloride intoxicated rabbit liver (a) Serum total
cholesterol, (b) Serum triacylglycerol, (c) Serum LDL cholesterol, (d) Serum
HDL cholesterol, (e) Total Non-HDL cholesterol and (f) LDL/HDL cholesterol
ratio |
Effect of ethanol extracts of C. odorata on LDL/HDL cholesterol ratio
in carbon tetrachloride induced liver damage: Result obtained (Fig.
2f) showed that intoxication of rabbits with carbon tetrachloride produced
an elevated LDL/HDL (2.159±0.194) which was significantly (p<0.05)
higher than the normal controls (0.553±0.050) and the ethanol extract
of C. odorata treated control (0.798±0.072). Treatment with ethanol
extract of C. odorata significantly (p<0.05) lowered LDL/HDL ratio
(1.001±0.090) in the intoxicated animals.
DISCUSSION
In this study, rabbits treated with single dose of CCl4 developed
a significant hepatic damage which manifested as a substantial increase in the
activities of serum marker enzymes, ALT, AST (Alqasoumi
et al., 2009), LDH and γ-GT. This was indicative of cellular
leakage and loss of functional integrity of cell membrane in liver (Mukherjee,
2003, 2002). Hepatotoxicity induced by Carbon tetrachloride,
impaired function of the hepatic cells in this study. Impairment of hepatic
function is evidenced by decreased serum protein and albumin concentration and
increased bilirubin concentration (Table 1). This is in agreement
with previous reports in which liver damage impaired function and decreased
protein and albumin concentrations in serum (Alisi et
al., 2008). In this study, increases in ALT, AST, LDH and γ-GT
activities showed damage to the tissues while elevations in total bilirubin,
decreases in total protein and albumin concentrations showed impairment of hepatic
functions. Most circulating proteins are synthesized in the liver and concentrations
indicate synthetic ability of the liver (Deepak et al.,
2000).
Liver injury resulting from carbon tetrachloride toxicity was followed by a
concomitant induction of abnormal lipoprotein phenotype. Increase in cholesterol,
triacylglycerol, low density lipoprotein cholesterol, total non-high density
lipoprotein cholesterol, LDL/HDL-ratio and decrease in HDL-cholesterol concentrations
were indicative of abnormal lipoprotein phenotype. The results obtained from
this study indicated that ethanol extracts of C.odorata was hypolipidemic.
The extract was able to normalize lipoprotein phenotype (HDL, LDL, LDL/HDL-ratio
and total non-HDL-cholesterol) which was altered by the induction of hepatotoxicity
with carbon tetrachloride in the rabbits. In the presence of Carbon tetrachloride-intoxication,
hypertriglyceridaemia resulted. In this condition, there is increased VLDL concentration
because of the action of hepatic lipase. The HDL becomes overloaded with triglyceride;
they reduce in size, losing apoA1 and the concentration of HDL-cholesterol
falls. This fall in HDL-cholesterol concentration represents an alteration of
the lipoprotein phenotype. Effectiveness of C. odorata extracts in the
normalization of lipoprotein phenotype in serum (Fig. 2a-f)
was seen in the restoration of lipid profile values to their respective normal
values. Normalization of lipid profile values could be a mechanism of/or a consequence
of hepatoprotection. More generally, normalization of lipoprotein phenotype
by the extracts could point to their potential to reduce cardiovascular disease
risk. Hypercholesterolaemic serum increases the permeability of endothelial
cells through zonula occludens-1 with phosphatidylinositol 3-kinase signaling
pathway (Chang et al., 2009). Increased endothelial
cell permeability will result in loss of endothelial relaxing factor and increased
vascular complication. Inhibition of the processes resulting to hypercholesterolemia
and by implication increased endothelial cell permeability by C. odorata
extracts may be a mechanism of reduction of vascular complication. It is well
known that total cholesterol, triglyceride, LDL-cholesterol, HDL-cholesterol,
LDL/HDL-ratio and total non-HDL cholesterol are all independent and significant
predictors of cardiovascular disease risk (Wilson et al.,
1998). The reduction in the total-non-HDL-cholesterol is most interesting
since it is the single greatest predictor of cardiovascular risk. Measurement
of total-non-HDL-cholesterol has been shown to be as good as or better than
apo-lipoprotein fractions in the prediction of cardiovascular risk. There has
been an association between hypolipemic potential and hepatoprotective effect
(Alisi et al., 2008). C. odorata ethanol
extract, may be furthering its hepatoprotective effect by hypolipidemic mechanism.
Chromolaena odorata normalises lipoprotein phenotypes in carbon tetrachloride
toxicity-induced dyslipidemia.
CONCLUSION In conclusion, present investigation revealed that C. odorata leaf extract has got the ability to prevent dyslipidemia that would normally result from Carbon tetrachloride-induced oxidative damage. The mechanism of this action is not fully understood. However, the protection conferred on the liver by the extract would have preserved the liver cell to function in the maintenance of lipoprotein phenotype. Intake of C. odorata extract as drug or as supplement in diet may offer useful benefit in the preservation of lipoprotein phenotype. This will also be beneficial in the reduction of cardiovascular risk associated with dyslipidemia.
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REFERENCES |
1: Achudume, A.C. and K.E. Ogunyemi, 2007. Effects of the extracts of Pycanthus angolensis against chemically induced acute hepatotoxicity. Pak. J. Biol. Sci., 10: 3231-3233. CrossRef | PubMed | Direct Link |
2: Adinarayana, K., K.N. Jayaveera, P.M. Rao, C.M. Chetty, D.K. Sandeep, C. Swetha and T.S.M. Saleem, 2011. Acute toxicity and hepatoprotective effect of methanolic extract of Tephrosia calophylla. Res. J. Med. Plant, 5: 266-273. CrossRef | Direct Link |
3: Alisi, C.S., A.A. Emejulu, P.N.C. Alisi, L.A. Nwaogu and O.O. Onyema, 2008. Decreased cardiovascular risk and resistance to hyperlipemia-induced hepatic damage in rats by aqueous extract of Urtica dioica. Afr. J. Biochm. Res., 2: 102-106. Direct Link |
4: Alisi, C.S. and G.O.C. Onyeze, 2008. Nitric oxide scavenging ability of ethyl acetate fration of methanol leaf extracts of Chromolaena odorata (Linn.). Afr. J. Biochem. Res., 2: 145-150. Direct Link |
5: Alisi, C.S. and G.O.C. Onyeze, 2009. Biochemical mechanisms of wound healing using extracts of Chromolaena odorata-linn. Nig. J. Bioch Mol. Biol., 24: 22-29.
6: Alqasoumi, S.I., M.S. Al-Dosari, A.M. AlSheikh and M.S. Abdel-Kader, 2009. Evaluation of the hepatoprotective effect of Fumaria parviflora and Momordica balsamina from Saudi folk medicine against experimentally induced liver injury in rats. Res. J. Med. Plant, 3: 9-15. CrossRef | Direct Link |
7: Austin, M.A., J.E. Hokanson and K.L. Edwards, 1998. Hypertriglyceridemia as a cardiovascular risk fator. Am. J. Cardiol., 81: 7B-12B. PubMed |
8: Babu, B.H., B.S. Shylesh and J. Padikkala, 2001. Antioxidant and hepatoprotective effect of Acanthus ilicifolius. Fitoterapia, 72: 272-277. CrossRef | Direct Link |
9: Baigent, C., A. Keech, P.M. Kearney, L. Blackwell and G. Buck et al., 2005. Efficacy and safety of cholesterol-lowering treatment: Prospective meta-analysis of data from 90 056 participants in 14 randomised trials of statins. Lancet, 366: 1267-1278. CrossRef | Direct Link |
10: Chang, B., X. Geng, W. Jianan, M. Ji, X. MeiXiang and C. Peng, 2009. . Hypercholesterolaemic serum increases the permeability of endothelial cells through zonula occludens-1 with phosphatidylinositol 3-kinase signaling pathway. J. Biomed. Biotech., 2009: 1-5. CrossRef |
11: Chavda, R., K.R. Vadalia and R. Gokani, 2010. Hepatoprotective and antioxidant activity of root bark of Calotropis procera R. Br (Asclepediaceae). Int. J. Pharmacol., 6: 937-943. CrossRef | Direct Link |
12: Dahm, J.L. and P.D. Jones, 1996. Mechanisms of Chemically Induced Liver Disease. In: Hepatology A Textbook of Liver Disease, Zakim, D. and T.D. Boyer (Eds.). WB Saunders, Pennsylvania, USA., pp: 875-890
13: Gopal, D.V. and H.R. Rosen, 2000. Abnormal findings on liver fucntion tests: Interpretating result to narrow the diagnosis and establish a prognosis. Postgrad. Med., 107: 100-114. CrossRef | PubMed |
14: Doumas, B.T., W.A. Watson and H.G. Biggs, 1971. Albumin standards and the measurement of serum albumin with bromcresol green. Clin. Chim. Acta, 31: 87-96. CrossRef | PubMed | Direct Link |
15: Gole, M.K., S. Dasgupta, R.K. Sur and J. Ghosal, 1997. Hepatoprotective effect of Amorra rohituka. Int. J. Pharm., 35: 318-322.
16: Gornall, A.G., C.J. Bardawill and M.M. David, 1949. Determination of serum proteins by means of the biuret reaction. J. Biol. Chem., 177: 751-766. PubMed | Direct Link |
17: Yen, G.C., H.H. Lai and H.Y. Chou, 2001. Nitric oxide scavenging and antioxidant effects of Uraria crinitia root. Food Chem., 74: 471-478. CrossRef |
18: Houghton, P.J., 1995. The role of plants in traditional medicine and current therapy. J. Altern. Complement. Med., 1: 131-143. CrossRef | Direct Link |
19: Irobi, O.N., 1997. Antibacterial properties of ethanol extract of Chromolaena odorata (Astreriaceae). Pharaceutical Biol., 35: 111-115.
20: Khadr, M.E., K.A. Mahdy, K.A. El-Shamy, F.A. Morsy, S.R. El-Zayat and A.A. Abd-Allah, 2007. Antioxidant activity and hepatoprotective potentials of black seed, honey and silymarin on experimental liver injury induced by CCl4 in rat. J. Applied Sci., 7: 3909-3917. CrossRef |
21: Koneru, A., S. Satyanarayana, K. Mukkanti and K.A. Khan, 2011. In vitro antioxidant activity of Itrifal Kishneezi: A unani formulation. Am. J. Drug Discovery Dev., 1: 121-128.
22: Gupta, A.K., H. Chitme, S.K. Dass and N. Misra, 2006. Antioxidant activity of Chamomile recutita capitula methanolic extracts against CCl4-induced liver injury in rats. J. Pharmacol. Toxicol., 1: 101-107. CrossRef | Direct Link |
23: Mayes, P.A. and K.M. Botham, 2003. Lipid Transport and Storage. In: Harpers Illustrated Biochemistry, Murray, R.K., D.K. Granner, P.A. Mayes and V.W. Rodwell (Eds.). 26th Edn., MacGraw-Hill, New York, pp: 205-218
24: Milionis, H.J., A.I. Kakafika, S.G. Tsouli, V.G. Athyros, E.T. Bairaktari, K.I. Seferiadis and M.S. Elisaf, 2004. Effects of statin treatment on uric acid homeostasis in patients with primary hyperlipidemia. Am. Heart J., 148: 635-640. PubMed |
25: Moundipa, P.F., S. Ngouela, G.A. Tchamba, N.F. Njayou, P.D.D. Chuisseu, F. Zelefack and E. Tsamo, 2007. Antihepatotoxic activity of Xylopia phloiodora extracts on some experimental models of liver injury in rats. Int. J. Pharmacol., 3: 74-79. CrossRef | Direct Link |
26: Mukai, M., K. Ozasa, K. Hayashi and K. Kawai, 2002. Varoius S-GOT/S-GPT ratios in nonviral liver disorders and related physical conditions and lifestyle. Dig. Dis. Sci., 47: 549-555. CrossRef |
27: Mukherjee, P.K., 2002. Quality Control of Herbal Drugs. 1st Edn., Business Horizons, New Delhi, pp: 546-549
28: Mukherjee, P.K., 2003. Plant Products with Hypercholesterolemic Potentials. In: Advance in Food and Nutrition Research, Steve, L. (Ed.). Elsevier Science, USA., pp: 277-338
29: Noori, S., N. Rehman, M. Qureshi and T. Mahboob, 2009. Reduction of carbon tetrachloride-induced rat liver injury by coffee and green tea. Pak. J. Nutr., 8: 452-458. CrossRef | Direct Link |
30: Owoyele, V.B., J.O. Adediji and A.O. Soladoye, 2006. Anti-inflammatory activity of aqueous leaf extract of Chromolaena odorata. Inflammopharmacology, 13: 479-484. PubMed |
31: Pearlman, P.C. and R.T. Lee, 1974. Detection and measurement of total bilirubin in serum, with use of surfactants as solubilising agents. Clin. Chem., 20: 447-453. PubMed | Direct Link |
32: Pessayre, D., A. Berson, B. Fromenty and A. Mansouri, 2001. Mitochondria in steatohepatitis. Semin Liver Dis., 21: 57-69.
33: Raja, S., K.F. Ahamed, V. Kumar, K. Mukherjee, A. Bandyopadhyay and P.K. Mukherjee, 2007. Antioxidant effect of Cytisus scoparius against carbon tetrachloride treated liver injury in rats. J. Ethnopharmacol., 109: 41-47. CrossRef | PubMed | Direct Link |
34: Rao, G.M.M., C.V. Rao, P. Pushpangadan and A. Shirwaikar, 2006. Hepatoprotective effects of rubiadin, a major constituent of Rubia cordifolia Linn. J. Ethnopharmacol., 103: 484-490. CrossRef | Direct Link |
35: Rao, K.S., P.K. Chaudhury and A. Pradhan, 2010. Evaluation of anti-oxidant activities and total phenolic content of Chromolaena odorata. Food Chem. Toxicol., 48: 729-732. CrossRef |
36: Singh, S.K., U. Dimri, M. Kataria and P. Kumari, 2011. Ameliorative activity of Withania somnifera root extract on paraquat-induced oxidative stress in mice. J. Pharmacol. Toxicol., 6: 433-439. CrossRef | Direct Link |
37: Weber, L.W.D., M. Boll and A. Stampfl, 2003. Hepatotoxicity and mechanism of action of haloalkanes: Carbon tetrachloride as a toxicological model. Crit. Rev. Toxicol., 33: 105-136. CrossRef | PubMed | Direct Link |
38: Lee, W.M., 2003. Drug-induced hepatotoxicity. New Eng. J. Med., 349: 474-485. CrossRef | Direct Link |
39: Wilson, P.W., K.M. Anderson and W.P. Castelli, 1991. Twelve-year incidence of coronary heart disease in middle-aged adults during the era of hypertensive therapy: The Framingham offspring study. Am. J. Med., 90: 11-16. PubMed |
40: Wilson, P.W., R.B. D'Agostino, D. Levy, A.M. Belanger, H. Silbershatz and W.B. Kannel, 1998. Prediction of coronary heart disease using risk factor categories. Circulation, 97: 1837-1847. CrossRef | PubMed | Direct Link |
|
|
|
 |