Hepatic endogenous defense potential of propolis after mercury intoxication

© 2008 ISZS, Blackwell Publishing and IOZ/CAS

ORIGINAL ARTICLE

Hepatic endogenous defense potential of propolis after mercury

intoxication

Monika BHADAURIA,1 Sangeeta SHUKLA,1 Ramesh MATHUR,1 Om Prakash AGRAWAL,2 Sadhana

SHRIVASTAVA,1 Sonia JOHRI,3 Deepmala JOSHI,1 Varsha SINGH,1 Deepak MITTAL1 and Satendra

Kumar NIRALA1

1Reproductive Biology and Toxicology Laboratory and 2Insect Physiology and Biochemistry, School of Studies in Zoology, Jiwaji

University, Gwalior, India, and 3Boston College for Professional Studies, Gwalior, India

Abstract

Exposure to mercuric chloride (HgCl2; 5 mg kg–1 body weight; i.p.) induced oxidative stress in mice and substantially

increased lipid peroxidation (LPO) and oxidized glutathione (GSSG) levels, decreased the level of reduced glu-

tathione (GSH) and various antioxidant enzymes in liver and also increased the activities of liver marker enzymes in

serum. Therapy with propolis extract, a resinous wax-like beehive product (200 mg kg–1 orally, after mercury

administration), for 3 days inhibited LPO and the formation of GSSG and increased the level of GSH in the liver.

Release of serum transaminases, alkaline phosphatase, lactate dehydrogenase and ã-glutamyl transpeptidase were

significantly restored after propolis treatment. The activities of antioxidant enzymes, that is, superoxide dismutase,

catalase, glutathione-S-transferase and glucose-6-phosphate dehydrogenase, were also concomitantly restored

towards normal levels after propolis administration. These observations clearly demonstrate that propolis treatment

augments antioxidant defense against mercury-induced toxicity and provide evidence that propolis has therapeutic

potential as a hepatoprotective agent.

Key words: antioxidant enzymes, glutathione, hepatoprotection, mercuric chloride, propolis.

Correspondence: Monika Bhadauria, Reproductive Biology and

Toxicology Laboratory, School of Studies in Zoology, Jiwaji

University, Gwalior 474011, India.

Email: [email protected]

INTRODUCTION

Over the past century, there has been an increasing

awareness of the health and developmental risks associ-

ated with environmental exposure to toxic metals, such as

lead (Pb), mercury (Hg), cadmium (Cd) and arsenic (As).

Although exposure to toxic levels of any of these envi-

ronmental contaminants can impair the health of adults,

the toxicological effects of these metals are often more

devastating in the developing central nervous system and

general physiological systems of children. Although Pb is

perhaps the most publicized and well known of the pediat-

ric metal intoxicants, Hg is at least equally toxic (Counter

& Buchanan 2004). The element Hg is classified as a heavy

metal (atomic weight 200.59) and exists in three forms: el-

emental Hg, inorganic Hg compounds (primarily mercuric

chloride) and organic Hg (primarily methylmercury). Mer-

cury is presented in different industrial settings, in the air,

as well as in drinking water and food, resulting in continu-

ous exposure to the entire human population and other

organisms (World Health Organization 1991).

Integrative Zoology 2008; 4: 311–321 doi: 10.1111/j.1749-4877.2008.00103.x


The use of elemental Hg or inorganic Hg salts in vari-

ous industrial procedures (Kark 1994) as well as organic

(alkyl and aryl) Hg compounds applied as disinfectants

and fungicides can be the source of its occupational

exposure. On a population level, Hg released into the en-

vironment and dental amalgam are major exposure sources

(Papp et al. 2005). The Hg++ form of inorganic Hg has a

great affinity for sulphydril (SH) groups of endogenous

biomolecules (Clarkson 1997) and is invariably found in

cells and tissues attached to thiol-containing proteins and

small-molecular-weight thiols, such as cysteine and re-

duced glutathione (GSH). In addition, Hg can also give

rise to free radicals that induce lipid, protein and DNA

oxidation (Lund et al. 1993; Clarkson 1997; Perottoni et al.

2004). Thus, inorganic Hg has been considered in the

present study. Therapy using natural products that might

suppress the formation of free radicals or neutralize/re-

move them from the body would be useful in reducing Hg

poisoning.

Apitherapy or therapy with bee products (e.g. honey,

pollen, propolis, fortified honey, herb honey) is an old

tradition that has been revived by recent researchers.

These products, which are used as health foods and

medicines, are receiving renewed focus on their beneficial

effects in a general “back to nature” trend (Dobrowolski

et al. 1991). Propolis is a resinous material collected by

honey bees from plant exudates, which is used for the

construction and repair of honey comb. It has a pleasant

aromatic odor and is yellow-green to dark brown in color,

depending on its source and age (Ghisalberti 1979). Pro-

polis has attracted much attention in recent years as a

useful substance for medicines and cosmetics, although

it has been used in folk medicine since ancient times

(Sforcin et al. 2000). Several empirical and clinical findings

point to the fact that propolis may be more effective against

pathogenic microorganisms than conventional

medications, with the added advantage that it causes mi-

nor side-effects (Miyaves et al. 1988), which is true of

many natural prescriptions (Higashi & Castro 1994). Pro-

polis has a long history of being used in traditional medi-

cine dating back to at least 300 BC (Ghisalberti 1979) and

has been reported to have a broad spectrum of biological

activities, including arthritis (Park & Kahng 1999; Hu et

al. 2005) and as an hepatoprotective agent against galac-

tosamine (Sugimoto et al. 1999), econazole (Liu et al. 2004),

tert-butyl hydroperoxide (Wang et al. 2006), paracetamol

(Seo et al. 2003; Nirala & Bhadauria in press), ethanol

(Sharma et al. 1997) and carbon tetrachloride (Bhadauria

et al. 2007a). Synergism between propolis and antibiotics

(Krol et al. 1993) and antibacterial agents (Stepanovic et

Figure 1 Therapeutic influence of propolis on mercury-induced alterations in (A) aspartate aminotransferase and (B) alanine

aminotransferase. P-value Hg versus control taking +P ≤ 0.05 and *P ≤ 0.01; P-value treatments versus Hg taking ++P ≤ 0.05 and**P ≤ 0.01. F = 62.9 and F = 57.8 for aspartate aminotransferase and alanine aminotransferase, respectively. C, control; P per

se, propolis per se; Hg, mercury; Hg+P, mercury + propolis; Hg+S, mercury + silymarin.

M. Bhadauria et al.


al. 2003) and with chelators against light-metal (Nirala et

al. 2008) and heavy-metal intoxication (Geckil et al. 2005)

has also been observed. In the present study, an attempt

has been made to assess the therapeutic potential of pro-

polis to improve the health status against Hg-induced tox-

icity through various marker enzymes, oxidative stress and

antioxidant status.

MATERIALS AND METHODS

Animals and chemicals

Female swiss albino mice (25 ± 5 g body weight, 8–10

weeks old) were maintained in the institutional animal fa-

Figure 2 Therapeutic influence of propolis on mercury-induced alterations in liver marker enzymes (A) serum alkaline phosphatase

(SALP), (B) lactate dehydrogenase (LDH) and (C) g-glutamyl transpeptidase ( -GT). P-value Hg versus control taking +P ≤ 0.05 and*P ≤ 0.01; P-value treatments versus Hg taking ++P ≤ 0.05 and **P ≤ 0.01. F = 176.3, F = 209.3 and F = 153.8 for SALP, LDH and -

GT, respectively. C, control; P per se, propolis per se; Hg, mercury; Hg+P, mercury + propolis; Hg+S, mercury + silymarin.

Propolis reverses mercury-induced toxicity


cility under standard husbandry conditions of light (14 h)

and dark (10 h) at a temperature of 25 ± 2°C and a relative

humidity of 60–70%. The animals were fed dry pellets con-

sisting of a standard animal diet (provided by the animal

facility) and given drinking water ad libitum. The experi-

mental protocols were approved and carried out accord-

ing to the guidelines set by the Institutional Animal Ethics

Committee.

Mercury, in the form of mercury chloride [HgCl2] and

silymarin was purchased from Sigma-Aldrich., St. Louis,

USA and crude propolis was collected from the hive of

Apis mellifera. All other chemicals used in the present

Figure 3 Therapeutic influence of propolis on mercury-induced alterations in (A) lipid peroxidation (LPO), (B) reduced glutathione

(GSH) and (C) oxidized glutathione (GSSG) in the liver. P-value Hg versus control taking +P ≤ 0.05 and *P ≤ 0.01; P-value treatments

versus Hg taking ++P ≤ 0.05 and **P ≤ 0.01. F = 124.2, F = 41.3 and F =135 for LPO, GSH and GSSG, respectively. C, control; P per

se, propolis per se; Hg, mercury; Hg+P, mercury + propolis; Hg+S, mercury + silymarin.

M. Bhadauria et al.


study were of the highest purity and analytical reagent

grade.

Preparation and administration of the doses

Mercuric chloride was dissolved in triple-distilled wa-

ter (5 mg kg–1) and administered intraperitoneally (i.p.)

(Sener et al. 2007). A series of extractions was carried out

to yield an ethanolic extract of propolis (62.8% w/w) as

described previously and the propolis was kept at 4°C

until use (Shukla et al. 2005). Aqueous suspensions of

propolis (200 mg kg–1, p.o.) and silymarin (50 mg kg–1, p.o.)

were prepared in 1% gum acacia suspension (GAS)

Figure 4 Therapeutic influence of propolis on mercury-induced alterations in antioxidant enzymatic status. (A) Glutathione-S-

transferase (GST), (B) glucose-6-phosphatase dehydrogenase (G6PDH), (C) super oxide dismutase (SOD) and (D) catalase (CAT).

P-value Hg versus control taking +P ≤ 0.05 and *P ≤ 0.01; P-value treatments versus Hg taking ++P ≤ 0.05 and **P ≤ 0.01. F = 15.6,

F = 17.6, F = 21.2 and F = 34.3 for GST, G6PDH, SOD and CAT, respectively. C, control; P per se, propolis per se; Hg, mercury;

Hg+P, mercury + propolis; Hg+S, mercury + silymarin.



(Bhadauria et al. 2007b) and silymarin was given as a posi-

tive control. The dose of propolis was selected on the

basis of our previous study (Shukla et al. 2005).

Experimental protocol

Thirty mice were assigned into five groups of six ani-

mals each. Group 1: received saline (i.p.; 0.9% NaCl) fol-

Figure 5 (A) Photomicrograph of a con-

trol liver showing well-arranged hepato-

cytes with well-formed intact nucleus,

obvious sinusoidal space (arrow) and

well-formed central vein (CV) (100×). (B)

Propolis per se treated liver showing

well-formed sinusoidal space (arrow)

with central vein (CV) (100×). (C) He-

patic injury induced by mercury show-

ing congestion in the sinusoidal space

with damaged central vein (CV) (100×).

(D) Propolis recovered the hepatocytes

with better cord arrangement and nor-

mal sinusoidal space (arrows) and well-

formed central vein (CV) (100×); (E)

Silymarin treatment shows normal hepa-

tocytes with prominent nucleus and

proper sinusoidal space (arrow) and nor-

mal central vein (CV) (100×).

M. Bhadauria et al.


lowed by GAS (5 mL kg–1, p.o.) for 3 days. Group 2: re-

ceived saline (as in group 1) followed by propolis per se

(200 mg kg–1, p.o.) for 3 days. Groups 3–5: received mercu-

ric chloride (5 mg kg–1 i.p., single dose). Group 3: served as

the experimental control and received GAS (5 mL kg–1, p.

o.) for 3 days after toxicant administration. Groups 4 and 5:

treated with propolis and silymarin, respectively, for 3 con-

secutive days after Hg intoxication.

Blood biochemical analysis

Twenty-four hours after the final administration the

animals were killed under a mild ether anesthesia and blood

was drawn immediately by puncturing the retro-orbital

venous sinus for isolation of serum and used to determine

the leakage of aspartate aminotransferase (AST) and ala-

nine aminotransferase (ALT) (Reitman & Frankel 1957),

lactate dehydrogenase (LDH) (Wroblewski & La Due 1955)

and serum alkaline phosphatase (SALP) (Halk et al. 1954).

ã-Glutamyl transpeptidase (ã-GT) was measured using the

Merck kit (Merck Specialties Private Limited, Mumbai,

India) as per the manufacturer’s instructions.

Oxidative stress and antioxidant status in the

liver

Tissue samples of the liver were homogenized with ice-

cold 1.15% KCl for the determination of thiobarbituric acid

reactive substances (TBARS) and the activities of glu-

cose-6-phosphate dehydrogenase (G6PDH), glutathione-

S-transferase (GST) and catalase (CAT). For the estima-

tion of GSH and superoxide dismutase (SOD) activity, the

tissues were homogenized in 5% sulfo-salicylic acid and

normal saline solution, respectively.

Hepatic LPO was measured by estimating TBARS as

described by Sharma and Krishna Murti (1968). Reduced

glutathione and oxidized glutathione (GSSG) were mea-

sured using the method of Roberts and Francetic (1993).

Total GST activity was measured according to the method

of Habig et al. (1974). Total SOD activity was estimated by

measuring the inhibition of auto oxidation of epinephrine

as per Mishra and Fridovich (1972), and CAT activity was

estimated using the method of Aebi (1984). The activity of

G6PDH was measured using the method of Askar et al.

(1996). Total protein was estimated using the method of

Lowry et al. (1951) with bovine serum albumin as standard.

Histopathological analysis

For photomicroscopic observations, liver samples were

fixed in Bouin’s fixative and routinely embedded in

paraffin. Tissue sections (5-ìm thickness) were stained with

hematoxylin–eosin (HE) and examined under a

photomicroscope.

Statistical analysis

The results are expressed as the mean ± standard error

(SE) of the six animals in each group. The data were sub-

jected to statistical analysis using a one-way ANOVA and

statistical significance was set a priori at P = 0.05. The

data were also subjected to a Student’s t-test with statis-

tical significance set a priori at P = 0.01 and P = 0.05; P-

value Hg versus control taking +=0.05 and *£0.01; P-value

treatments versus Hg taking ++=0.05 and **=0.01 for

Student’s t-test (Snedecor & Cochran 1994).

RESULTS

Blood biochemical analysis

Figure 1 (a,b) and Fig. 2 (a–c) represent the effect of

propolis extract on Hg-induced blood biochemical

alterations. The AST, ALT, SALP, g-GT and LDH activi-

ties were significantly increased after Hg intoxication (P =

0.01). Propolis therapy significantly recovered all the en-

zymes back toward the control level (P = 0.01) and showed

>70% protection in all parameters except g-GT, which

showed more than 60% protection in its release. Mercury

administration significantly increased serum bilirubin con-

tents (P = 0.01), which were significantly diminished by

propolis (P = 0.01) and showed more than 80 % protection.

Oxidative stress and antioxidant status

The protective effect of propolis on the Hg-induced

experimentally oxidative stress and antioxidant status are

shown in Figs 3 (a–c) and 4 (a–d). Hepatic LPO increased

with Hg administration (P = 0.01). The administration of

propolis and silymarin significantly inhibited LPO (P £ 0.

01) and reduced hepatic peroxidative stress by more than

80% (Fig. 3a). Acute administration of Hg caused a signifi-

cant decrease in hepatic GSH contents and increased GSSG

contents (P = 0.01). Propolis therapy recovered the GSH

contents towards the control (P = 0.01) and showed better

protection compared with the positive control (Fig. 3b).

Three days of treatment with propolis diminished the con-

tents of GSSG (P = 0.01) and the ANOVA was significant at

P = 0.05 (Fig. 3c). Mercury diminished the antioxidative

status of the organs after its administration and decreased

the activities of GST (Fig. 4a), G6PDH (Fig. 4b), SOD (Fig.

4c) and CAT (Fig. 4d) (P = 0.01). Treatment with propolis

and silymarin recovered the activities of all these enzymes

towards normal levels (P = 0.01). Recovery of 60% was

found in SOD and CAT, whereas more than 70% improve-

ment was observed in the activities of GST and G6PDH



with propolis treatment.

Histopathological observations

Figure 5 (A–E) presents the histology of the liver after

the different treatments. Light microscopic evaluation

showed regular morphology of the liver parenchyma with

well-designed hepatic cells and sinusoids in the control

group and with propolis per se (Fig. 5A,B). The Hg-treated

group showed severe liver injury with prominent conges-

tion in the sinusoidal space and a damaged central vein

(Fig. 5C). The hepatocytes were found with normal

appearance, arranged in cord in most of the areas having

prominent nucleus and granular cytoplasm after propolis

(Fig. 5D) and silymarin treatment (Fig. 5E).

DISCUSSION

Acute administration of Hg caused toxic effects in the

liver and this damage was observed to be associated with

increases in LPO and GSSG, indicating oxidative tissue

damage, and with decreases in antioxidant enzymatic ac-

tivities and a significant reduction in GSH levels. Treat-

ment of the animals with propolis extract and silymarin

appeared to afford protection against this noxious stimulus.

Because of the high affinity of Hg to thiol groups, Hg is

known to affect living organisms by damaging thiol pro-

teins and enzymes (Clarkson 1997). Divalent ions of tran-

sition metals, especially Pb, Hg and copper, can promote

LPO and much attention is currently focused on LPO in

the pathogenesis of metal toxicity. The LPO is an auto-

catalytic free-radical process whereby polyunsaturated

fatty acids in cell membranes undergo degradation by a

chain reaction to yield lipid hydroperoxides, which subse-

quently decompose to form a variety of products, includ-

ing malondialdehyde (MDA).

The LPO induction by Hg has been demonstrated in

the present study and cell membrane permeability might

be affected by this process (Nava et al. 2000; Mahboob et

al. 2001). Propolis treatment was found to protect the liver

against Hg toxicity and suppressed the process of LPO.

Antioxidants have a protective effect against tissue inju-

ries in the pathogenesis of which LPO may be involved

and previous studies have demonstrated that quercetin (a

major component of propolis) inhibits LPO effectively by

scavenging free radicals and/or transition metal ions (Terao

& Piskula 1999). Flavonoids and phenolics are a group of

naturally occurring compounds that are widely distributed

as secondary metabolites in the plant kingdom and pre-

vent oxidative injury and cell death by protecting against

LPO (Molina et al. 2004) and by chelating the metals

(Afanasev et al. 1989).

The present study suggests that Hg administration

depletes GSH content and increases GSSG in the liver,

which makes hepatocytes more susceptible to oxidative

damage, particularly during increased free radical

production. The GSH is an important intracellular antioxi-

dant (hydrogen-donating compound) that spontaneously

neutralizes several electrophiles and reactive oxygen spe-

cies (Lu 1999); and the contents of GSH/GSSG play a key

role in cells and maintain the redox status of the cell (Rana

et al. 2002). Concomitant restoration of the GSH pool after

propolis treatment was consistent with some other reports

(Nirala et al. 2008). GSH is a substrate of the enzyme GST,

a phase II enzyme that plays a key role in the cellular

detoxification of xenobiotics, electrophiles and reactive

oxygen species through their conjugation to GSH (Mates

2000). In the present study, reduction in GST activity after

Hg exposure might decrease the conjugation reaction and

increase the toxic events. GSSG is reduced back to GSH

by nicotinamide adinine dinucleotide phosphate

(NADPH)-dependent reaction. G6PDH, the first enzyme

of the pentose phosphate pathway, is an important site of

NADPH production. Reduction in the activity of G6PDH

enzyme after Hg might affect the rate of generation of

NADPH and, thus, affect the redox status of the cell. Pro-

polis therapy maintained the normal status of GST and

G6PDH activities and, thus, improved the antioxidant sta-

tus of the hepatocytes in terms of increased GSH and de-

creased GSSG contents. Previous studies have shown that

propolis prevents hepatic disorders induced by acetami-

nophen (Nirala & Bhadauria 2008) and CCl4 (Bhadauria et

al. 2007a; b) as well as light-metal-induced lipid

peroxidation in liver homogenates (Nirala et al. 2008) ow-

ing to the presence of flavonoids and phenolics.

Mercury-induced toxic effects arise from alterations in

the structural integrity of the mitochondrial inner

membrane, resulting in a loss of normal cation selectivity,

which allows the membrane to participate effectively in

oxidative metabolism (Lund et al. 1993). Mercury ions per-

turb the mitochondrial inner membrane function, which

results in depletion of mitochondrial GSH content and in-

creased formation of H2O2 by the mitochondrial electron

transport chain (Lund et al. 1991). Increased H2O2 forma-

tion might be accompanied by increased peroxidation of

mitochondrial lipids, which is consistent with an oxidative

stress-like condition (Lund et al. 1993). Cells have a num-

ber of mechanisms to protect themselves from the toxic

effects of reactive oxygen species. The SOD removes su-

per oxide (O2) by converting it to H

2O2, which can be rap-

idly converted to water by CAT. The activity of SOD is a

M. Bhadauria et al.


sensitive index to hepatic damage because it scavenges

the superoxide anion to form hydrogen peroxide, dimin-

ishing the toxic effect. CAT is an enzymatic antioxidant

widely distributed in all animal tissues that decomposes

hydrogen peroxide and protects the tissue from highly

reactive hydroxyl radicals. Therefore, alteration in the ac-

tivity of these two enzymes might result in a number of

deleterious effects because of the accumulation of super-

oxide radicals and hydrogen peroxide. In the present study,

propolis extract significantly restored SOD and CAT

activity, which indicated that propolis extract could scav-

enge reactive free radicals and eventually lessen oxida-

tive damage to the tissues and subsequently improve the

activities of these antioxidant enzymes. It is proposed that

the beneficial effects of a pharmacological dose of propo-

lis might result from its antioxidant properties by reviving

the endogenous antioxidant defense system.

Mercury intoxication resulted in significant hepatic

damage and oxidative stress as evidenced by substantial

increases in the serum activities of AST, ALT, SALP, LDH

and g-GT into the circulation as a result of hepatocellular

nacrosis, which causes an increase in the permeability of

the cell membrane (Sharma et al. 2002). A reduction in the

levels of these parameters toward the respective normal

values by propolis therapy is an indication of the stabili-

zation of the plasma membranes and reparation of the he-

patic tissue damage caused by Hg. This effect is in agree-

ment with the commonly accepted view that serum levels

of transaminases would return to normal after healing of

the hepatic parenchyma and the regeneration of

hepatocytes. Histopathological observations of the im-

provement in the histoarchitecture of the liver after pro-

polis treatment are also consistent with the biochemical

findings of the present study.

In conclusion, our findings demonstrate that propolis

extract is able to reverse pathological parameters, such as

necrotic histological features, and serum levels of AST,

ALT, SALP, LDH and g-GT of acute liver damage induced

by Hg. This protective ability of propolis resulted from its

modulatory effects on detoxification (GST and G6PDH)

and antioxidative enzymes (SOD and CAT), which, in turn,

suppress the production of free radicals and reduce sub-

sequent liver damage. Furthermore, the high content of

polyphenols and flavonoids in propolis contributes to free

radical scavenging and antioxidation activities. Therefore,

dietary propolis administration might be useful as a

hepatoprotective agent against Hg-induced acute liver

damage.

ACKNOWLEDGMENTS

The authors wish to thank the Indian Council of Medi-

cal Research, New Delhi, India (45/15/2002/PHA/BMS) for

financial assistance.

REFERENCES

Aebi HL (1984). Catalase in vitro. Method of Enzymol-

ogy 105, 121–6.

Afanasev IB, Dorozhko AI, Brodskii AV, Kostyuk VA,

Potapovitch AI (1989). Chelating and free radical

scavenging mechanisms of inhibitory action of rutin

and quercetin in lipid peroxidation. Biochemical

Pharmacology 38, 1763–9.

Askar MA, Sumathy K, Baquer NJ (1996). Regulation

and properties of glucose-6-phosphate dehydroge-

nase from rat brain. Indian Journal of Biochemistry

and Biophysics 33, 512–18.

Bhadauria M, Nirala SK, Shukla S (2007a). Propolis pro-

tects CYP2E1 enzymatic activities and oxidative stress

induced by carbon tetrachloride. Molecular and

Cellular Biochemistry 302, 215–24.

Bhadauria M, Nirala SK, Shukla S (2007b). Duration

dependent hepatoprotective effect of propolis extract

against carbon tetrachloride induced acute damage

in rats. Advances in Therapy 24, 1134–43.

Clarkson TW (1997). The toxicology of mercury. Criti-

cal Reviews in Clinical Laboratory Sciences 34,

369–403.

Countera SA, Buchanan LH (2004). Mercury exposure

in children: a review. Toxicology and Applied Phar-

macology 198, 209–30.

Dobrowolski JW, Vohora SB, Sharma K, Shah SA, Naqvi

SAH, Dandiya PC (1991). Antibacterial, antifungal,

antiamoebic, anti-inflammatory and antipyretic stud-

i e s on p ropo l i s bee p roduc t s . Journa l o f

Ethnopharmacology 35, 77–82.

Geckil I, Ates B, Durmaz G, Erdogan S, Yilmaz I (2005).

Antioxidant, free radical scavenging and metal

characterictics of propolis. American Journal of Bio-

chemistry and Biotechnology 1, 27–31.

Ghisalberti E (1979). Propolis: a review. Bee World 60,

59–84.

Halk PB, Oster BL, Summerson WH (1954). The Practi-

cal Physiological Chemistry, 14th edn. McGraw Hill

Book Company, New York.

Habig WH, Pabst MT, Jakoby WB (1974). Glutathione–

Stransferases. The first enzymatic step in mercaptu-



ric acid formation. Journal of Biological Chemistry

249, 7130–39.

Higashi KO, de Castro SL (1994). Propolis extracts are

effective against Trypanosoma curzi and have an

impact on its interaction with host cells. Journal of


Hu F, Hepburn HR, Li Y, Chen M, Radloff SE, Daya S

(2005). Effects of ethanol and water extracts of pro-

polis (bee glue) on acute inflammatory animal models.

Journal of Ethnopharmacology. 100, 276–83.

Kark RAP (1994). Clinical and neurochemical aspects of

inorganic mercury intoxication. In: de Wolff FA, ed.

Handbook of Clinical Neurology, 20. Elsevier,

Amsterdam, pp. 367–411.

Krol W, Scheller S, Shani J, Pietsz G, Czuba Z (1993).

Synergistic effect of ethanolic extract of propolis and

antibiotics on the growth of Staphylococcus aureus.

Arzneim-Forsch. 43: 607–9.

Liu CF, Lin CH, Lin CC, Lin YH, Chen CF, Lin CK, Lin SC

(2004). Antioxidative natural product protect against

econazole induced liver injuries. Toxicology 196, 87–

93.

Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951).

Protein measurement with Folin’s phenol reagent.

Journal of Biological Chemistry 193, 265–75.

Lu SC (1999). Regulation of hepatic glutathione

synthesis: current concepts and controversies. The

FASEB Journal 13, 1169–83.

Lund BO, Miller DM, Woods JS (1991). Mercury-in-

duced H2O2 production and lipid peroxidation and

lipid oxidation in vitro in rat kidney mitochondria.

Biochemical Pharmacology 42, 181–7.

Lund BO, Miller DM, Woods JS (1993). Studies on Hg

(II)-induced H2O2 formation and oxidative stress in

vivo and in vitro in rat kidney mitochondria. Bio-

chemical Pharmacology 45, 2017–24.

Mahboob M, Shireen KF, Atkinson A, Khan AT (2001).

Lipid peroxidation and antioxidant enzyme activity

indifferent organs of mice exposed to low level of

mercury. Journal of Environmental Science and

Health B 36, 687–97.

Mates JM (2000). Effects of antioxidant enzymes in the

molecular control of reactive oxygen species

toxicology. Toxicology 153, 83–104.

Mishra P, Fridovich I (1972). The role of superoxide

anion in the autooxidation of epinephrine and a

simple assay for superoxide dismutase. Journal of

Biological Chemistry 247, 3170–75.

Miyares C, Hollands I, Castaneda C, Gonzales T, Fragoso

T, Curras R, Soria C (1988). Ensayoterapeutico con

un preparado a base de propoleo propolisina en la

giardiasis del humano. Acta Gastroenterologia

Latina-Americana 18, 195–201.

Molina MF, Sanchez-Reus I, Iglesias I, Benedi J (2003).

Quercitin, a flavonoids antioxidant, prevents and

protects against ethanol induced oxidative stress in

mouse liver. Biological and Pharmaceutical Bulle-

tin 26, 1398–1402.

Nava M, Romero F, Quiroz Y, Parra G, Bonet L,

Rodriquez-Iturbe B (2000). Melatonin attenuation of

acute renal failure and oxidative stress induced by

mercuric chloride in rats. American Journal of Physi-

ology for Renal Physiology 279, F910–18.

Nirala SK, Bhadauria M (2008). Propolis reverses ac-

etaminophen induced hepatorenal biochemical and

histopathological alterations. Archives of Pharmacal

Research 31, 451-461.

Nirala SK, Bhadauria M, Mathur R, Mathur A (2008).

Influence of á-tocopherol, propolis and piperine on

therapeutic potential of tiferron against beryllium

induced toxic manifestations. Journal of Applied

Toxicology 28, 44–54.

Papp A, Nagymajtenyi L, Vezer T (2005). Subchronic

mercury treatment of rats in different phases of

ontogenesis: functional effects on the central and

peripheral nervous system. Food Chemistry and

Toxicology 43, 77–85.

Park EH, Kahng JH (1999). Suppressive effects of pro-

polis in rat adjuvant arthritis. Archives of Pharamcal

Research 22, 554–8.

Perottoni J, Lobato LP, Silveira A, Rocha JBT, Emanuelli

T (2004). Effects of mercury and selenite on d-

aminolevulinate dehydratase activity and on selected

oxidative stress parameters in rats. Environmental

Research 95, 166–73.

Rana SV, Allen T, Singh R (2002). Inevitable glutathione,

then and now, Indian Journal of Experimental Bi-

ology 40, 706–16.

Reitman S, Frankel S (1957). A colorimetric method for

determination of serum glutamic oxaloacetic and

glutamic pyruvic transaminases. American Journal

of Clinical Pathology 28, 56–63.

Roberts JC, Francetic DJ (1993). The importance of

sample preparation and storage of glutathione

analysis. Analytical Biochemistry 211, 183–7.

Sener G, Sehirli O, Tozan A, Ovunc AV, Gedik N, Omurtag

M. Bhadauria et al.


GZ (2007). Ginkgo biloba extract protects against

mercury(II)-induced oxidative tissue damage in rats.

Food and Chemical Toxicology 45, 543–50.

Seo KW, Park M, Song YJ, Kim SJ, Yoon KR (2003). The

protective effects of propolis on hepatic injury and

its mechanism. Phytotherapy Research 17, 250–53.

Sforcin JM, Fernandes JA, Lopes CAM, Bankova V,

Funari SRC (2000). Seasonal effect on Brazalian pro-

p o l i s a n t i b a c t e r i a l a c t i v i t y . J o u r n a l o f


Sharma M, Pillai KK, Husain SZ, Giri DK (1997). Protec-

tive role of propolis against alcohol-carbon tetrachlo-

ride-induced hepatotoxicity in rats. Indian Journal

of Pharmacology 29, 76–81.

Sharma MK, Kumar M, Kumar A (2002). Ocimum sanc-

tum aqueous leaf extract provides protection against

mercury induced toxicity in Swiss albino mice. In-

dian Journal of Experimental Biology 40, 1079–82.

Sharma SK, Krishna Murti CR (1968). Production of lipid

peroxides by brain. Journal of Neurochemistry 15,

147–9.

Shukla S, Bhadauria M, Jadon A (2005). Evaluation of

hepatoprotective potential of propolis extract in car-

bon tetrachloride induced liver injury in rats. Indian

Journal of Biochemistry and Biophysics 42, 321–5.

Snedecor GW, Cochran WG (1994). Statistical Method,

8th edn. Iowa State University Press, Ames.

Stepanovic S, Antic N, Dakic I, Svabic-Vlahovic M

(2003). In vitro antimicrobial activity of propolis and

synergism between propolis and antimicrobial drugs.

Microbiology Research 158, 353–7.

Sugimoto Y, Tarumi T, Kaneko Y et al. (1999). Effect of

propolis extract on D-galactosamine-induced hepatic

injury in rats. Biological and Pharmaceutical Bul-

letin 22, 1237–9.

Terao J, Piskula MK (1999). Flavonoids and membrane

lipid peroxidatiotn inhibition. Nutrition 15, 790–91.

Wang BJ, Lien YH, Su CL, Wu CP, Yu ZR (2006). Frac-

tionation using supercritical CO2 influences and

hepatoprotective activity of propolis against induced

by tert-butyl hydroperoxide. International Journal

of Food Science and Technology 41, 68–75.

World Health Organization (1991). Inorganic mercury.

Environmental Health Criteria 118. World Health

Organization, Geneva, pp. 1–168.

Wroblewski F, La Due JS (1955). Lactic dehydrogenase

activity in blood. Proceedings of the Society for Ex-

perimental Biology and Medicine 90, 210–13.


Hepatic endogenous defense potential of propolis after mercury intoxication

Documents

Transcript of Hepatic endogenous defense potential of propolis after mercury intoxication