0.5 MLKG
0.5 MLKG
0.5 MLKG
Received: July 23, 2012, accepted: July 27, 2012, published: August 20, 2012
Original article:
* corresponding author: Department of Biology, Science and Research Branch, Islamic Azad
University, P.O. Box 16535-446, Tehran, Iran, E-mail: [email protected],
[email protected], Tel: +98 9123380064, Fax: +9821 44865939
ABSTRACT
The inner bark of cinnamon (Cinnamomum zeylanicum L.) is commonly used as a spice and
has also been widely employed in the treatment and prevention of disease. The aim of the pre-
sent study is to evaluate the protective effect of cinnamon bark extract against carbon tetra-
chloride (CCl4)-induced liver damage in male Wistar rats. Administration with cinnamon ex-
tracts (0.01, 0.05 and 0.1 g/kg) for 28 days significantly reduced the impact of CCl4 toxicity
on the serum markers of liver damage, aspartate aminotransferase, alanine aminotransferase
and alkaline phosphatase. In addition, treatment of cinnamon extract resulted in markedly in-
creased the levels of superoxide dismutase and catalase enzymes in rats. The histopathologi-
cal studies in the liver of rats also supported that cinnamon extract markedly reduced the tox-
icity of CCl4 and preserved the histoarchitecture of the liver tissue to near normal. Thus, the
results suggest that cinnamon extract acts as a potent hepatoprotective agent against CCl4 in-
duced hepatotoxicity in rats.
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EXCLI Journal 2012;11:495-507 – ISSN 1611-2156
Received: July 23, 2012, accepted: July 27, 2012, published: August 20, 2012
limited and there is a great demand for the (CAT) were purchased from Parsazmoon
development of new effective drugs. A Company of Iran. All other reagents used in
number of studies have shown that the plant the experiment were of analytical grade.
extracts having antioxidant activity protect
against CCl4 hepatotoxicity by inhibiting Preparation of the plant powder and
lipid peroxidation and enhancing antioxi- extraction
dant enzyme activity (Shahjahan et al., The plant materials were obtained from
2004; Sheweita et al., 2001). the local market. Shade dried cinnamon
Cinnamon (Cinnamomum zeylanicum bark was milled and extracted using ethanol
L., Lauraceae) is a tropical evergreen tree 80 % in Soxhlet apparatus for 8 h. Then, the
and grows wild in Sri Lanka, Madagascar, extract was evaporated to dryness and the
India and Indochina. The inner bark of the final dry extract was stored in dark
tree has been used in ethno-medicine and at -20 °C until used for the experiments.
flavoring for foods (Bakkali et al., 2008; The percentage yield of extract was 17.7 %
Baytop, 1999). In addition to its culinary (w/w) of the initial raw material.
uses, cinnamon has been employed in tradi-
tional herbal medicine to treat a variety of Phytochemical analysis
health conditions (Gruenwald et al., 2010). The qualitative phytochemical analysis
Some studies showed that extracts and its of the crude extract of Cinnamomum
constituents from cinnamon also posses an- zeylanicum L. bark was carried out to de-
timicrobial (Carmo et al., 2008; Chao et al., termine the active phytochemical constitu-
2000; Dusan et al., 2006; Ranasinghe et al., ents which were responsible for the hepato-
2002; Shahverdi et al., 2007), insecticidal protective activity. Some of these methods
(Yang et al. 2005), acaricidal (Fichi et al., were as follows:
2007), antityrosinase (Marongiu et al., 20 mg extract was dissolved in 10 ml
2007), antioxidant and antimutagenic (Ja- ethanol and filtered. 0.5 ml conc. HCl and
yaprakasha et al., 2007) activities. In addi- magnesium ribbon were added to 2 ml fil-
tion, other evidence suggests that cinnamon trate. Development of pink-tomato red col-
may be effective in the treatment of cancer or indicated the presence of flavonoids (Pa-
(Hyeon et al., 2003; Nishida et al., 2003) rekh et al., 2006). 20 mg extract was dis-
and infectious diseases (Hayashi et al., solved in 2 ml distilled water and filtered.
2007; Premanathan et al., 2000), and that it 2 ml FeCl3 was added to the filtrate, blue-
also shows anti-inflammatory (Hong et al., black precipitate indicated the presence of
2002; Tung et al., 2008), antioxidant (Su et tannins (Parekh et al., 2006). To 0.5 ml of
al., 2007; Murcia et al., 2004; Okawa et al., the filtrate obtained in alkaloids test 5 ml
2001), hypotensive (Preuss et al., 2006), distilled water was added. Frothing persis-
and cholesterol-lowering effects (Khan et tence indicated the presence of saponins
al., 2003; Subash Babu et al., 2007). The (Parekh et al., 2006). 20 mg extract was
present study investigates the hepatoprotec- dissolved in 2 ml distilled water and fil-
tive potential of cinnamon ethanolic extract tered. To the filtrate, 2-4 drops of 1 % HCl
treatment against CCl4-induced liver toxici- was added and steam was passed through it.
ty in rats. To the 1 ml of this solution 6 drops of
Wagner's reagent was added. Brownish-red
MATERIALS AND METHODS precipitate indicated the presence of alka-
loids (Finar, 2003). Salkovski test was per-
Chemicals
CCl4 was obtained from Merck, Germa- formed using a small amount of extract so-
ny. Assay kits for alanine aminotransferase lution. To this solution 5 drops of conc.
(ALT), aspartate aminotransferase (AST), H2SO4 and 1 ml chloroform were added.
alkaline phosphatase (ALP), total protein, Change of yellow color into red indicated
superoxide dismutase (SOD) and catalase the presence of terpenoids (Finar, 2003). A
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Table 1: Body weight, liver weight and liver index of acute CCl4 (50% CCl4/olive oil)-treated rats with or without cinnamon ethanolic extract
Initial body weight (g) 224 ± 10.4 216 ± 9.8 223 ± 13.2 219 ± 12.5 221 ± 11.2 218 ± 8.9 220 ± 11.4
Final body weight (g) 256 ± 15.3 249 ± 17.2 226 ± 10.5 +++ 232 ± 8.5 239 ± 13.6 240 ± 17.4 * 245 ± 15.2 **
Weight gain (g) 32 ± 4.9 33 ± 3.7 3 ± 0.7 +++ 13 ± 3.4 18 ± 6.3 * 22 ± 5.9 * 25 ± 4.1 **
Liver weight (g) 5.3 ± 0.34 5.2 ± 0.41 8.5 ± 1.2 +++ 6.9 ± 0.64 6.5 ± 0.71 ** 6.3 ± 0.57 ** 5.9 ± 0.83 ***
Liver index 2.07 ± 0.04 2.09 ± 0.03 3.76 ± 0.07 +++ 2.97 ± 0.05 2.72 ± 0.04 ** 2.63 ± 0.03 ** 2.41 ±0.03 ***
Values are mean ± S.E.M. (n = 9 rats per each group). The liver index was calculated as liver weight / body weight × 100, * p <0.05, ** p<0.01, ***,
p<0.001 significantly different from the group treated with CC14. +++. p<0.001 significantly different from the control
Table 2: Effects of cinnamon ethanolic extract on serum and liver biochemical indices in CCl4-induced hepatotoxicity in rats
Parameters Control Cinnamon extract CCl4 CCl4 + Cinnamon extract (g/kg b.w.)
(0.05 g/kg b.w.) 0.005 0.01 0.05 0.1
ALT (IU/L) 44.81 ± 3.28 54.00 ± 4.71 595.33± 18.04 449.28 ± 67.45 266.00 ± 14.91 358.66 ± 28.83 417.40 ± 44.28
+++ *** *** *
AST (IU/L) 125.81 ± 10.28 197.25 ± 5.20 671.80 ± 19.63 572.71 ± 30.34 331.66 ± 20.60 335.83 ± 38.10 512.00 ± 23.15
+++ *** *** **
ALP (IU/L) 280.45 ± 13.44 270.75 ± 15.28 559.00 ± 17.17 593.00 ± 22.89 384.16 ± 25.36 420.00 ± 26.35 401.71 ± 10.37
+++ *** *** ***
SOD (U/mg protein) 9.4 ± 1.7 9.1 ± 2.1 5.8 ± 1.3 6.4 ± 0.9 6.9 ± 1.5 7.6 ± 1.3 8.3 ± 1.1
+++ * ** ***
CAT (U/mg protein) 4.05 ± 0.05 3.75 ± 0.05 0.61 ± 0.01 3.15 ± 0.05 2.45 ± 0.05 2.55 ± 0.05 0.95 ± 0.15
+++ *** *** ***
Total protein (g/dL) 62.66 ± 2.6 57.66 ± 3.57 26.27 ± 0.81 35.50 ± 1.84 41.52 ± 1.83 43.07 ± 3.38 45.01 ± 4.52
+++ *** *** ***
Values are mean ± S.E.M. (n= 9 rats per each group). * p <0.05, ** p<0.01, *** p<0.001 significantly different from the group treated with CC14. +++.
p<0.001 significantly different from the control
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Control 0 0 0 0
Cinnamon extract (0.05 g/kg b.w.) 0 0 0 0
CCl4 4 3 3 3
Cinnamon extract
0.005 g/kg b.w. + CCl4 2 1 2 1
0.01 g/kg b.w. + CCl4 0 0 0 1
0.05 g/kg b.w. + CCl4 3 2 2 2
0.1 g/kg b.w. + CCl4 2 2 2 2
a
Livers were scored for hepatic injury via light microscopy with score 0 = no visible cell damage; score
1 = focal hepatocyte damage on less than 25 % of the tissue; score 2 = focal hepatocyte damage on
25-50 % of the tissue; score 3 = extensive, but focal, hepatocyte lesions; score 4 = global hepatocyte
necrosis.
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Figure 1A: Liver section of normal control Figure 1D: Liver section from rat treated with
showing normal central vein (arrow) and radiat- 0.005 g/kg b.w. + CCl4 shows fatty degenera-
ing hepatocytes (arrowhead) (H&E*16) tion (arrowhead), necrosis and infiltration of in-
flammatory cells (arrow) (H&E*640)
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