Antigout and Antioxidant Activity of Stem and Root of Origanum majorana Linn.
Surendra Kumar Sharma
The present study was aimed at investigating the antioxidant and antigout activity of the various extracts of the stem and root of Origanum majorana Linn. The antioxidant activity of ethanol extracts of both stem and root of marjoram was evaluated by various in vitro antioxidant assays. The ethanol extracts of stem and root showed potent antioxidant activity in all models when compared with ascorbic acid having IC50 values 21.05 μg mL-1 and 84.98 μg mL-1 for DPPH (2, 2-diphenyl-picrylhydrazyl); 492.8 μg mL-1 and 477.6 μg mL-1 for H2O2 radical scavenging assay; 156.9 and 141.79 μg mL-1 for metal chelating assay, respectively. The reducing ability of root ethanol extract (absorbance 0.747±0.23) was found to be high, compared to stem ethanol extract (0.527±0.14). The in vitro xanthine oxidase inhibitory activity was performed on ethanol and aqueous extracts (stem and root) of the plant and activity was found to be significant with IC50 59.21 and 148 μg mL-1 in both ethanol extracts. Further in-vivo antigout was studied gout induced in rats. Oral administration of stem and root ethanol extracts (200 and 400 mg kg-1 body weight) showed a significant decrease in uric acid, Creatinine, ESR (Erythrocyte Sedimentation Rate) and MDA (Malondialdehyde) levels in the gouty rats. A significant increase was observed in reduced glutathione. No change in protein content was noticed. In conclusion, the studied plant extracts showed significantly variable anti-gout activity associated with both antioxidant and anti-inflammatory effects which may be due to the presence of flavonoids, phenolics, saponins and triterpenoidal compounds revealed by preliminary phytochemical screening.
Received: November 20, 2013;
Accepted: March 11, 2014;
Published: April 12, 2014
Gout is a multi-factorial disease affecting the flexibility of joints. It is
usually characterized by re-current attacks of acute inflammatory arthritis-a
red, tender, hot, swollen joints leading to bursitis. It is caused by elevated
uric acid levels in the blood, resulting into deposition of urate (as monosodium
urate-monohydrate) crystals, generally known as tophi crystals in joints, tendons
and surrounding tissues, characterized by hyper-uricemia and in chronic stage,
may lead to renal failure (Singh et al., 2010).
These crystals cause an acute inflammatory response and can induce a permanent
tissue damage which is characterised by the appearance of ulceration of the
joint cartilage, marginal osteophytosis, geodic and erosive lesions and chronic
inflammation of synovial membrane (Dalbeth and Haskard,
2005; Corrado et al., 2006).
Current treatments to gout includes (a) NSAIDS such as ibuprofen, naproxen,
indomethacin, aspirin, etoricoxib (cox-2 selective inhibitors); corticosteroids
such as prednisone; allopurinol, probencid, colchicines (to decrease severity
of episodes). Although these agents are generally effective, they generates
superoxide (Berry and Hare, 2004) and lead to several
side effects such as skin allergies, fever, rash and diarrhoea progressively
developing leukocytosis, eosinophilia, vasculitis, aseptic meningitis, nephritis
and renal dysfunction, and hepatic dysfunction (Nguyen et
al., 2004; Strazzullo and Puig, 2007). So, there
is a need of herbal extracts with antioxidant property to nullify oxidative
and inflammatory response produced by xanthine oxidase.
Origanum majorana Linn. is a tender perennial aromatic herb of
the mint family (Labiatae), rich in phenols, flavonoids and terpenoids (Novak
et al., 2002; Raina and Negi, 2012; Proestos
and Komaitis, 2006; Janicsak et al., 1999).
Polyphenols (Costantino et al., 1992), flavonoids
(Chang et al., 1993; Selloum
et al., 2001), coumarins (Chang and Chiang,
1995), ellagic acid, Valoneic Acid Dilactone (VAD) (Unno
et al., 2004) have been reported as potent plant-based Xanthine Oxidase
The objective of the present research was to determine the potential of stem
and root of Origanum majorana as a potent antioxidant and antigout agent.
This was accomplished through testing different extracts of stem and root of
the plant as uric acid lowering (XOI), in vitro antioxidant and in
vivo anti-inflammatory in experimental gout model in rats. The possible
role of marjoram in reducing serum uric acid levels and its effect on biomarkers
of oxidative stress was also investigated. In the present investigation XO inhibitory
activity of O. majorana was evaluated to discover a natural substitute
of plant origin, which could have a superior effect of inhibiting XO activity
and can be used as an alternative to allopurinol for the treatment of gout as
well as for the treatment of other inflammatory-related diseases.
MATERIAL AND METHODS
Collection and authentication of plant material: Plant was collected from
Guru Jambheshwar University of Science and Technology Hisar in August, 2012.
Identified by Dr. K.C Bhatt, Senior Scientist, National Herbarium of Cultivated
Plants, National Bureau of Plant Genetic Resources, New Delhi, vide reference
no. NHCP/NBPGR/2012-28/. The plant has been deposited in Pharmacognosy division
of Department of Pharmaceutical Sciences, GJUS&T, Hisar for further references.
Chemicals: Folin-Ciocalteu reagent, 2, 2-Diphenyl-Picrylhydrazyl (DPPH),
hydrochloric acid (HCl), ferric chloride (FeCl3) and gallic acid
were obtained from Sigma-Aldrich, USA. Sodium carbonate (Na2CO3),
methanol, potassium dihydrogen phosphate (KH2PO4), sodium
chloride (NaCl), sodium hydroxide and hydrochloric acid were purchased from
S D Fine Chemicals Ltd., New Delhi. Xanthine, xanthine oxidase of bovine milk
origin, allopurinol (standard drug) and potassium oxonate (gout inducing drug)
were obtained from Sigma Aldrich, USA. All the other chemicals used were of
analytical grade, obtained from S D Fine Chemicals Ltd., New Delhi.
Animals: Swiss albino rats were procured from disease free small animal
house of LLRUVS, Hisar (Haryana). The experimental protocol was approved vide
letter no. IAEC/136-144 dated 10.01.13 by Institutional Animals Ethics Committee
(IAEC) and animal care was taken as per the guidelines of Committee for the
Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Govt.
of India (Registration No. 0436). Rats were maintained at normal pellet diet
in the animal house of Guru Jambheshwar University of Science and Technology,
Preparation of the extracts: The 1000 g powdered herb (stem and root)
of Origanum majorana was extracted successively first with 2.5 L of ethanol
(95 %) by using soxhlet apparatus and then with 2.5 L of water by cold maceration
for 72 h. The extracts were filtered and concentrated at 45°C using rotary
vacuum evaporator. The extract obtained was vacuum dried and stored for further
Preliminary phytochemical screening: The ethanol and aqueous extracts
of both stem and root of were screened for the presence of alkaloids, glycosides,
carbohydrates, sterols, phenolic compounds, tannins, flavonoids, saponins, proteins
and free amino acids using standard procedures (Goyal et
al., 1997; Kar, 2003; Kokate
et al., 2005; Ahmad, 2007).
In vitro antioxidant activity
DPPH radical scavenging assay: Plant extracts (stem and root ethanol)
and standard ascorbic acid solution (1 mL) of different concentrations viz;
25, 50, 75, 100, 125 and 150 μg mL-1 were added to 3 mL of a
0.004% methanol solution of DPPH. An equal amount of methanol and DPPH served
as control. After 30 min incubation in the dark, absorbance was recorded at
517 nm and the percentage inhibition was calculated. All the tests were performed
in triplicate and the graph was plotted with the average of three observations
(Kumaran and Karunakaran, 2007). The percentage inhibition
was calculated from:
where, Ao is the absorbance of the control and A1 is the absorbance
of the extract/standard.
Scavenging of hydrogen peroxide: A solution of hydrogen peroxide (20
mM) was prepared in phosphate buffer saline (pH 7.4), different concentrations
of plant extracts and standard ascorbic acid solution viz; 10, 25, 50, 100,
200 and 500 μg mL-1 in methanol (1 mL) were added to hydrogen
peroxide solution (2 mL). Absorbance of hydrogen peroxide at 230 nm was determined
after 10 min against a blank solution containing phosphate buffer without hydrogen
peroxide. A separate control sample was prepared for back ground subtraction.
The percentage inhibition activity was calculated with the average of three
observations (Kumaran and Karunakaran, 2007; Srinivasan
et al., 2007):
where, Ao is the absorbance of the control and A1 is the absorbance
of the extract/standard.
Ferric reducing power assay: Different concentrations of plant extracts
and standard ascorbic acid solution viz; 10, 25, 50, 100, 200 and 500 μg
mL-1 in 1 mL of methanol were mixed with phosphate buffer (2.5 mL,
0.2 M pH 6.6) and potassium ferricyanide (2.5 mL, 1%). The mixture was incubated
at 50°C for 20 min. A portion (2.5 mL) of tricholoroacetic acid (10%) was
added to the mixture which was then centrifuged at 3,000 g (rpm) for 10 min
at room temperature. The upper layer (2.5 mL) was mixed with distilled water
(2.5 mL) and ferric chloride (0.5 mL, 0.1%) and the absorbance of the reaction
mixture indicated increased reducing power. The absorbance was measured at 700
nm. All the tests were performed in triplicate and the graph was plotted with
the average of three observations (Kumaran and Karunakaran,
2007; Kumar et al., 2005).
Metal chelating complex assay: The reaction mixture containing 1 mL
of different concentrations of extracts viz. 10, 20, 40, 80, 100 and 200 μg
mL-1 in methanol were added to 0.1 mL of 2 mM ferrous chloride and
0.2 mL of 5 mM ferrozine to initiate the reaction and the mixture was shaken
vigorously and left to stand at room temperature for 10 min. The absorbance
of the solution was measured at 562 nm. The positive controls were those using
ascorbic acid and all tests and analysis were run in triplicate (Subhashini
et al., 2011). The percentage chelating effect of ferrozine-Fe2+
complex formation was calculated from:
where, Ao is the absorbance of the control and A1 is the absorbance
of the extract/standard.
In-vitro xanthine oxidase inhibitory activity: Xanthine oxidase
inhibition assay was measured spectrophotometrically at λ 295 nm under
aerobic condition (Unno et al., 2004; Umamaheswari
et al., 2007). The reaction mixture contained solution of xanthine
as a substrate, a solution of enzyme (xanthine oxidase) and the test solutions
(50, 100, 250 and 500 μg mL-1) prepared in Di-Methyl Sulfoxide
(DMSO). Concentrations of allopurinol Viz., 10, 25, 50 and 100 μg mL-1
were prepared in phosphate buffer (ph 7.5). To each 1 mL sample, added 2.9 mL
phosphate buffer and 2 mL xanthine (0.15 Mm) solution. The sample mixture was
pre-incubated for 10 min at 30°C. 0.1 mL of xanthine oxidase (0.1 units
mL-1) was added to each test tube and incubated at 30°C for 30
min. After the incubation period, 1 mL of 1 N HCL was added to stop the reaction
and the contents homogenized. After then absorbance was measured at 284 nm to
see the formation of uric acid that occurs in the test solution and percentage
inhibition was determined according to the following equation:
IC50 values were obtained through linear regression analysis the
plot of concentration against percent inhibition.
In-vivo antigout activity
Experimental design: Thirty swiss rats were maintained on laboratory
stock diet and divided into five groups, each group comprised of 6 rats. Group1
(Normal control), received only vehicle (0.5% carboxy methyl cellulose in water)
whereas group 2 (Gouty control) received Potassium Oxonate (P.O) (250 mg kg-1
i.p dissolved in 0.9% saline) solution for the induction of gout along with
the vehicle (Yonetani et al., 1987). An hour
later group 3, 4, 5 (gouty rats) received root alcohol, stem alcohol and standard
drug (allopurinol) at the oral dose of 200, 400, 10 mg kg-1 extract,
respectively. The procedure was repeated for 4 days. Rats were fasted for 16
h before blood collection on fourth day. After 1 h from injection, blood samples
were withdrawn from eye vein orbital and divided into two parts one mixed with
sodium citrate (109 mmol L-1) for determination of erythrocyte sedimentation
rate (ESR) (Westergren, 1921; Gilmour
and Sykes, 1951) and the sec part mixed with heparin for the separation
of plasma for the determination of uric acid (Watts, 1974;
Yonetani et al., 1987), malondialdehyde (MDA)
as an indicator for lipid per-oxidation (Satoh, 1978),
glutathione (Ellman, 1959) and creatinine content (Owen
and Johns, 1999) as an indicator for renal functioning. Along with this,
animals were sacrificed and total protein present in the liver tissue was determined
by standard methods (Smith et al., 1985).
Statistical analysis: The results obtained were expressed as the mean±SEM.
Rats of the control gouty group were compared with the healthy rats. Test groups
were compared with the control gouty groups. The significance of values was
analysed by ANOVA followed by Dunnets
Test statistical value of p>0.5 was considered as significant.
The ethanol extract of both stem and root showed the presence of terpenoids,
flavonoids and tannins whereas saponins and carbohydrates were present in stem
and root aqueous extracts, respectively. Alkaloids, glycosides ad proteins were
absent in both of the extracts (root and stem).
In-vitro antioxidant activity: A significant decrease was observed
in the concentration of DPPH radicals due to the scavenging ability of extracts
of the plant. The activity was found to be dose dependent. Maximum scavenging
activity was observed at different concentrations in stem and root, respectively.
All the extracts demonstrated H+-donor activity. The highest DPPH
radical scavenging activity was detected in stem ethanol extract (IC50
21.05 μg mL-1) in comparison to root ethanol extract (84.98
μg mL-1) and that of standard ascorbic acid was (IC50
8.45 μg mL-1).
A significant dose dependent response in the hydrogen peroxide scavenging activity
aws reflected in both ethanol stem (IC50 492.8 μg mL-1)
and root ( IC50 477.6 μg mL-1) extracts whereas,
IC50 value of standard was found to be 173.4 μ mL-1.
The extracts of O. majorana interferes with the ferrous-ferrozine complex
and the red colour of the complex is decreased with the increasing concentrations
of the extracts. Both the extracts captured ferrous ions before ferrozine and
thus have ferrous chelating ability. The root ethanol extract showed the highest
ferrous ion chelating ability (IC50 141.79 μg mL-1)
among both the extracts (stem ethanol extract: IC50 156.9 μg
mL-1; standard: IC50 15.8 μg mL-1).
Figure 1 showed the reductive capabilities of different extracts
of O. majorana when compared to the standard, ascorbic acid. The root
ethanol extract showed the highest reducing ability (absorbance 0.747±0.23)
compared to stem ethanol extract (0.527±0.14).
Ferric reducing power assay. Absorbance
of standard (Std), stem ethanol extract (S.A), root ethanol extract (R.A)
at different concentrations
|| Xanthine oxidase (XO) inhibitory activity of allopurinol
Xanthine oxidase inhibitory activity.
std: Standard, S.A: Sstem ethanol extract, R.A: Root ethanol extract,
S.W: Stem water, RW: Root water, SH: Stem hydrosol, Ap.H: Aerial part
hydrosol, showing %inhibition at concentration 100 μg mL-1
In-vitro xanthine oxidase inhibitory activity: The XO inhibitory
activity of allopurinol and optimized different extracts of O. majorana
at various concentrations by in-vitro method are represented in Table
1. The percent xanthine oxidase inhibitory activity of the assayed samples
was determined through the slope of the plot of absorbance against time (sec).
IC50 values were obtained through the slope of plot of concentrations
(50, 100, 250, 500 μg m-1) against percent inhibition.
Ethanol extracts of stem and root has shown significant XO inhibition with
42.4 and 75.4% inhibition at concentration of 100 μg mL-1 whereas
aqueous extracts demonstrated non significant XO %inhibition with values of
25.62, 21.83% stem and root aqueous extract, respectively. The percentage inhibition
was found to be concentration dependent and comparable to the standard (78.0%)
in case of root ethanol extract as shown in Fig. 2.
The IC50 values of stem, root ethanol and aqueous extracts were
calculated from the curve and compared with standard (allopurinol IC50
1.11 μg mL-1). A significant value of IC50 i.e.148
and 59.21 was obtained in case of stem and root ethanol extracts, whereas very
high value of IC50 i.e., 500 and 600, respectively was determined
in stem and root aqueous extracts.
In-vivo anti-gout activity: In-vivo antigout activity
was determined for the extracts that showed significant results in in-vitro
anti-gout model and further dose of the extracts (stem ethanol extract: 400
mg kg-1; root ethanol extract: 200 mg kg-1) were decided
according to significant results.
Xanthine oxidase (XOD) catalyzes the oxidation of hypoxanthine to xanthine
and then to uric acid and is a key enzyme in the pathogenesis of hyperuricemia.
The ability of extracts of O. majorana to inhibit uricase was investigated
in this study.
|| Various biochemical parameters studied during in-vivo
|Values are presented as mean±S.E.M n: 6 in each group,
one way ANOVA followed by Dunnets test. ap<0.01 significant
as compared to normal control, *p<0.005, **p< 0.001 as compared to
Treatment with the uricase inhibitor potassium oxonate significantly affected
various bio-chemical parameters (uric acid, ESR, creatinine, MDA, glutathione,
total protein content) on wistar rats blood (Table 2).
An increased serum urate levels compared to normal control group was observed
after potassium oxonate injection. Allopurinol (10 mg kg-1), significantly
(p<0.01) reduce serum urate levels of hyperuricemic rats to values than that
found in normal animals. A four day treatment with ethanol extracts (stem and
root) at the dose of 200 mg kg-1 and 400 mg kg-1, respectively
reduce significantly (p<0.01) serum uric acid levels compared to hyperuricemic
The present study also investigated the efficacy of orally administered O.
majorana extracts and allopurinol on serum non-invasive biomarkers of oxidative
stress (malondialdehyde concentration) and lipid peroxidation (reduced glutathione)
levels in rats blood. As shown in Table 2 significant increase
(p<0.01) in serum was observed in root and stem ethanol extract treated normal
rats compared to positive control group in case of MDA levels, whereas a drastic
decrease in glutathione levels. A similar effect was obtained in hyperuricemic
animals following extracts administration compared to hyperuricemic control
In the present study gouty rats showed a significant increase in Erythrocyte
Sedimentation Rate (ESR) and creatinine levels. The oral administration of ethanol
root and stem extracts of marjoram produced the significant reduction in ESR
and creatinine levels 3 hours after i.p administration of oxonate. The root
and stem ethanol extract showed significant (p<0.01) decrease in ESR level
compared to positive gouty control rats and values are comparable to the standard
(allopurinol) whereas in case of reduction of creatinine levels, root extract
showed more significant (p<0.01)reduction than stem ethanol extract (p<0.05).
As shown in Table 2, the amount of protein content did not
vary neither with administration of oxonate nor with the oral dose of extracts.
There were insignificant (p>0.015) results shown by both stem and root ethanol
extracts compared to positive control in the total protein content in the liver
tissues of rats treated with the uricase inhibitor, potassium oxonate. It was
thus concluded that as per the results, there is no effect of potassium oxonate
on protein content in liver tissues.
DISCUSSION AND CONCLUSION
Anti-oxidant activity: Antioxidants exert their mode of action by suppressing
the formation of reactive oxygen species either by inhibition of enzymes or
by chelating trace elements. DPPH is widely used to evaluate the free radical
scavenging effect of natural antioxidants. DPPH is a stable free radical at
room temperature, it contains an odd electron which is responsible for visible
deep purple (Sreejayan and Rao, 1996). DPPH shows a
strong absorption band at 517 nm in visible spectrum. As the electron became
paired in the presence of free radical scavenging the absorption diminishes
and the resulting discoloration stochiometrically coincides with the number
of electrons taken up. The bleaching of DPPH absorption represents the capacity
of the test drugs to scavenge free radicals independently. The bleaching of
DPPH molecules can be correlated with the number of available hydroxyl groups.
A significant decrease was observed in the concentration of DPPH radicals in
all extracts of the plant. The highest DPPH radical scavenging activity was
detected in stem ethanol extract. It can be concluded that, the good antioxidant
activity of the extract may be probably due to the presence of substance with
an available hydroxyl group.
Hydroxyl radicals are the major active oxygen species that cause lipid oxidation
and enormous biological damage. Thus, scavenging of H2O2
is a measure of the antioxidant activity of the fractions. H2O2
can cross membranes and may oxidize a number of compounds. Both the ethanol
extracts of stem and root of O. majorana scavenged hydrogen peroxide
which may be attributed to the presence of phenolic groups that could donate
electrons to hydrogen peroxide, thereby neutralizing it into water (Haraguchi
et al., 2001). A dose dependent hydrogen peroxide scavenging activity
in both ethanol stem and root extracts was observed with IC50 values
492.8 and 477.6, respectively.
Ferrozine can quantitatively form complexes with Fe2+ but in the
presence of ion chelating agents, the complex formation is disrupted resulting
in a reduction in the red colour of the complex measurement of the rate of reduction
of the colour, therefore,s allows estimation of the chelating activity of the
coexisting chelator (Halliwell et al., 1987).
The absorbance of Fe2+-Ferrozine complex linearly decreased in dose
dependent manner. Both the extracts (stem and root ethanol) demonstrated an
effective capacity for iron binding, suggesting that its action as peroxidation
protector may be related to its iron binding capacity. In this assay the extract
and standard compound interfered with the formation of ferrous complex with
the reagent ferrozine, suggesting that it has chelating activity and captures
the ferrous ion before ferrozine. The results of our study express that the
extracts have an effective capacity for iron binding, suggesting its antioxidant
potential. In addition, the metal chelating ability of the fractions demonstrated
that they reduce the concentration of the catalysing transition metal involved
in the peroxidation of lipids. The root ethanol extract showed the highest ferrous
ion chelating ability (IC50; 141.79 μg mL-1) among
both the extracts.
The conversion of Fe3+ into Fe2+ in the presence of various
fractions was calculated to determine the reducing power ability (Umamaheswari
et al., 2007). The reducing ability of a compound generally depends
on the existence of reductones (antioxidants) which exert the antioxidant activity
by breaking the free radical chain by donating a hydrogen atom. The root ethanol
extract showed the highest reducing ability (absorbance 0.747±0.23) compared
to stem ethanol extract (0.527±0.14). The antioxidant principles present
in the extracts cause reducing power ability.
Based on the results obtained, it may be concluded that both ethanol extracts
(stem and root) of O. majorana showed strong antioxidant activity, reducing
power ability, free radical scavenging activity and metal chelating ability
when compared to standards such as ascorbic acid which may be due to presence
of flavonoids, phenols, tannins (phenolic compounds) and triterpenoids found
in the preliminary phytochemical screening. Overall, the plant extract is a
source of natural antioxidant that can be important in disease prevention, health
preservation and promotion of longevity.
Anti-gout activity: Gouty arthritis is characteristically acute inflammatory
reaction which occur in response to articular deposits of monosodium crystals
and associated with edema and erythema of joints, together with severe pain
(Sabina et al., 2010). Gout is an inherited
metabolic disease and results from hyperuricemia, an elevation of serum uric
acid level. Control of hyperuricemia is most often achieved by reducing uric
acid production with an inhibitor of XO, the enzyme catalyzing the two terminal
reactions in uric acid synthesis, or less frequently, by employing uricosuric
agents to increase renal clearance of uric acid (Boss and
Seegmiller, 1979; Emmerson, 1996). To date, the
only commercially available XO inhibitor is allopurinol, a purine analogue in
clinical use for more than 30 years (Takano et al.,
2005). However, allopurinol generates superoxide (Berry
and Hare, 2004) and lead to several side effects such as skin allergies,
fever, rash and diarrohea progressively developing renal dysfunction and hepatic
dysfunction (Osada et al., 1993; Takano
et al., 2005). The hypouricaemic activity was found to be significant
with IC50 59.21 and 148 in both root and stem ethanol extracts, after
in vitro studies, hence the further in-vivo study was performed.
The doses of extracts (root ethanol extract: 200 mg kg-1; stem ethanol
extract: 400 mg kg-1) given to the rats were selected based on the
results of in-vitro xanthine oxidase inhibitory activity. Serum uric
acid levels were elevated to approximately two-fold in oxonate-induced hyperuricaemic
rats. The groups treated with stem, root ethanol extracts and allopurinol for
four days decreased the uric acid level to almost normal value.
Flavonoids are a group of polyphenolic compounds which exhibit several biological
effects such as anti-inflammatory, anti-hepatotoxic, antiulcer activities, etc.
They are well known anti-oxidants and used as therapeutic agents for diseases
mediated by free radicals (Rice-Evans et al., 1996).
Also, the structureactivity relationship of different chemical classes
of flavonoids have been reported as potential inhibitors of XO (Costantino
et al., 1992). Since, phytochemical screening of the different extracts
of O. majorana revealed the presence of flavonoids, phenolics, saponins
and triterpenoids accounting for its antioxidant property, the present activity
of the extracts may be attributed to the same.
A significant increase (p<0.01) in serum was observed in root and stem ethanol
extract treated normal rats compared to positive control group in case of MDA
levels, whereas a decrement in levels of glutathione was there. Both the extracts
(root and stem ethanol) have shown significant decrease in level of MDA and
increase in glutathione level, comparable to normal. Thereby revealing that
serum uric acid through lipid peroxidation, might be working towards the etiopathogenesis
of oxidative stress diseases and its serum level may be a deciding factor for
progression of the disease. In conclusion, root and stem ethanol extracts (more
percentage was found in root extract) prevented oxidative stress by enhancing
total antioxidant capacity and decreasing lipid peroxidation. Moreover, the
hypouricemic and XO inhibitory action of ethanol extracts of O. majorana
has been confirmed in this study. Taking into account that marjoram is an antioxidant,
non toxic and can be used safely long term, this feature makes it a possible
alternative for allopurinol, or at least in combination therapy to minimise
the side-effects of allopurinol. Therefore, the use of suboptimal dosages of
allopurinol in combination with dietary changes may provide a safer approach
for prevention and treatment of hyperuricemia.
The authors are grateful to express our gratitude to Dr. K.C Bhatt, NBPGR,
Pusa campus, New Delhi for identification and authentification of the plant
for our project. We thank LLRUVS, Hisar (Haryana) for providing the animals
for biological studies.
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