Phytochemical and antiulcer properties of aqueous crude extract of Sargassum wightii from subtidal region of the Mandapam coast area, Gulf of ...
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Research Journal of Biotechnology Vol. 18 (4) April (2023)
Res. J. Biotech.
Phytochemical and antiulcer properties of aqueous
crude extract of Sargassum wightii from subtidal region
of the Mandapam coast area, Gulf of Mannar, India
Varshini R.1, Suganyadevi P.1* and Hariprasath L.2*
1. Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore, INDIA
2. Department of Biochemistry, School of Life Sciences (Ooty Off Campus), JSS Academy of Higher Education and Research, Mysuru, INDIA
*suganyadevi.palanisamy@kahedu.edu.in; hariprasath80@gmail.com
Abstract occurs because of an imbalance between aggressive
Peptic ulcers (PU) are sores or lesions in the injurious (e.g. pepsin, HCl) and defensive mucosa-protective
gastrointestinal mucosa extending throughout the factors (e.g. prostaglandins, mucus and bicarbonate barrier
muscularis mucosae, typically characterized by and adequate blood flow)37. All ulcers of the upper
gastrointestinal tract were originally thought to be caused by
different stages of necrosis, neutrophil infiltration,
the aggressive action of pepsin and gastric acid on mucosa.
blood flow reduction, increased oxidative stress and
inflammation. Peptic ulcer is the disease that is However, the denomination ―peptic ulcer23 has lately
associated with wound in the muscular mucous layer of pointed to Helicobacter pylori infection where the chronic
the stomach. Sargassum wightii is a brown seaweed uses of non-steroidal anti-inflammatory drugs (NSAIDs)
that contains bioactive components including fucoidan and acetylsalicylic acid (ASA) are some of the disease-
and fucoxanthin as well as large amounts of causing factors. The most common symptom of a peptic
polyphenols and is used by locals for the treatment of ulcer is burning abdominal pain that extends from the navel
peptic ulcer. to the chest, which can range from mild to severe. Other
common signs of a peptic ulcer include: changes in appetite,
nausea, bloody or dark stools (melena), unexplained weight
The objective of this study was to evaluate the
loss, indigestion and vomiting20. Maintaining a healthy
phytochemical and anti-ulcer properties of aqueous lifestyle through a balanced diet rich in fruits, vegetables and
crude extract of Sargassum wightii using a rat ulcer whole grains, quitting smoking and other tobacco use will
model. Antiulcer activity was performed by analysing also help in the prevention of peptic ulcer disease9.
the content of total protein, sialic acid, fucose, hexose,
glycoproetein, lipid peroxidation, catalase and In East Asian nations such as China, Korea and Japan,
superoxide dismutase enzymes in gastric juice and by seaweeds are employed both as therapeutic substances and
histopathology studies on stomach tissue after as nutritious food13. Seaweeds have attracted a lot of
treatment. The results showed that 500 mg/kg of the attention in recent years as a source of active component
seaweed extract showed good anti-ulcer activity as metabolites24. Brown seaweeds are generally attributed due
compared with the standard drug (20 mg/kg to the existence of the carotenoid fucoxanthin and the major
polysaccharides found include alginates, laminarins, fucans
Omeprazole). Thus, the present work validates the use
and cellulose10,12. Seaweeds are low in calories and high in
of Sargassum wightii for gastric ulcer treatment. soluble dietary fibers, proteins, minerals, vitamins,
antioxidants, phytochemicals and polyunsaturated fatty
Keywords: Sargassum wightii, Phytochemical, Acute acids16. Seaweed is a type of marine algae that is a wonderful
toxicity (LD50), Antiulcer activity. food source with several health advantages.
Introduction Sargassum wightii belongs to the family Sargassaceae, order
Peptic ulcers (PU) are sores or lesions in the gastrointestinal Fucales, subclass Cyclosporeae and class Phaeophyceae3,22.
mucosa extending throughout the muscular mucosae, It is a common coastal brown alga found on Indian
typically characterized by different stages of necrosis, shorelines. It is dark brown, 21-40 cm tall and densely
neutrophil infiltration, blood flow reduction, increased branching, with a spherical to ellipsoidal midrib 5-8 mm
oxidative stress and inflammation7. Peptic ulcers (PU) length and 2-4 mm broad23. These species are found in
manifest as a non-fatal disease, mainly represented by tropical and subtropical areas of the world and are known to
periodic symptoms of epigastric pain, which are often produce metabolites of various structural classes including
relieved by food or alkali. Ulcers trigger much discomfort to terpenoids, polysaccharides, polyphenols, sargaquinoic
patients, disrupting their daily routines and also causing acids, sargachromenol, plastoquinones, steroids and
mental agony14. The disease is mostly categorized based on glycerides, among others.
its anatomical origins such as gastric (found along the lesser
curvature of the stomach) and duodenal (occurring in the Fucoidan is a soluble polysaccharide with a wide range of
duodenal bulb—the most exposed area to gastric acid) biological actions including anticoagulant, antithrombotic,
ulcers38. Studies have shown that peptic ulcer disease (PUD) immunomodulatory, anticancer and antiviral properties.
https://doi.org/10.25303/1804rjbt064073 64
Research Journal of Biotechnology Vol. 18 (4) April (2023)
Res. J. Biotech.
This is a large, eukaryotic, aerobic, nonvascular, coastal with the animal house conditions (a cross-ventilated room
marine organism rich in sulfated polysaccharides with with temperature between 25°C and 28°C, 12 h light/12 h
significant free radical scavenging and antioxidant dark cycle) before the commencement of the study. The
properties, as well as hypolipidemic and anti-inflammatory research was conducted in accordance with the ethical
properties6,17,31. This study has led us to evaluate the committee guidelines and ethical clearance
phytochemical and anti-ulcer properties of aqueous crude (KAHE/IAEC/2021/11-09/007), the ARRIVE guidelines
extract of Sargassum wightii. (reporting of in vivo experiment) and the CPCSEA,
Government of India, guide for the CARE and use of
Material and Methods laboratory animals.
Seaweed Collection: Brown alga, Sargassum wightii
weighing about 1 kg was collected at the sub tidal region of Evaluation of ulcer activity using Wistar rats
the Mandapam coast (Lat. 09°17’ N; Long 79°07’ E, Gulf of Experimental design: The thirty (30) Wistar rat weighing
Mannar, India). The brown sea weeds were washed with 120 – 200g were divided into six groups of five rats each and
distilled water to remove salt, sand and epiphytes before fasted for 18 h before administration of extract. The rats in
being shade-dried in an air-conditioned chamber for 5 days. group 1 were considered as negative control treated as 1
The shade-dried specimen was chopped and powdered using ml/kg water. Group 2 was considered as positive control and
a mixer grinder. treated with omeprazole (20 mg/kg). Groups 3, 4 and 5 were
treated with aqueous crude extract of seaweed at 100 mg/kg,
Seaweed extracts: Crude extracts were obtained using 250 mg/kg and 500 mg/kg. Group 6 was treated with tween
water, the powdered sea weed samples were soaked with 80 (5ml/kg) and indomethacin was administrated to each rat
water at room temperature for 48 h and filtered through to induce ulcer after 30 min. The stomachs were dissected
Whatmann filter paper under vacuum; the filtrate was out and opened along the greater curvature, carefully rinsed
concentrated under negative pressure at 50⁰C using a rotary with distilled water, fixed in 10 % formalin and pinned on a
vacuum evaporator, Superfit, India. The crude extracts were cork board to examine the lesion with the help of hand lens.
stored at -20 ⁰ C until analysis. The stomachs were cut open, the stomach content was
deposited in a beaker, the ulcer index was determined.
Qualitative phytochemical analysis: The crude extract was
screened for the presence or absence of secondary The ulcer index computes the amount and time span for a
metabolites such as reducing sugars, alkaloids, steroidal percentage difference in relation to previous highs. As a
compounds, phenolic compounds, cardiac glycosides, result, the greater is the drawdown, the longer it will take for
flavonoids, saponins and tanninins using standard a stock to recover and return to its original high point and
procedures2. thus the higher will be the ulcer index. The number and
severity of the lesion were observed and scored8. To evaluate
Acute Toxicity Studies (LD50): The Lorke18 method of the antiulcerogenic activity, the status of lipid-peroxidation
LD50 determination was used, this method employs 13 (LPO), catalase (CAT), superoxide dismutase (SOD) and the
Wistar rats. The first phase is determination of the toxic glycoproteins (total hexoses, hexosamine, fucose and sialic
range. The rat was placed in three groups (n = 3) and will be acid) was estimated by appropriate methods.
given 10, 100 and 1000 mg/kg solubilized in a solution of 3
%, v/v Tween 85. The treated rat was observed for 24 h for The gastric content was collected in test tube and centrifuged
number of deaths. The death pattern in the first phase at 3000 rpm for 10 min. The pH of the supernatant was
determines the dose for the second phase. If no rats died measured using digital pH meter. The volume of supernatant
during the first phase, a new group of four rats will be was measured and expressed as ml/100 g body weight4,32.
provide 150, 250, or 500 mg/kg of the extract. The treated
animals was observed for lethality or signs of acute Determination of free and total acidity in gastric juice:
intoxication for 24 h. The LD50 is the geometric mean of the The free and total acidity of gastric juice collected from
highest non-lethal dose and the least toxic dose. The LD50 pylorus-ligated rats was determined by volumetric
was calculated by the formula: analysis15. One mL of gastric juice was pipetted into a 100
mL conical flask and 2 to 3 drops of Topfer’s reagent
LD50 = √(Do × D100) (HiMedia Pvt., Ltd., Mumbai) were added and titrated with
0.01N NaOH (which was previously standardized with
where Do = Highest dose that gave zero mortality dose and 0.01N oxalic acid) until all traces of red colour disappear and
D100 = Lowest dose that produced mortality. turn to yellowish orange.
Animal husbandry: Thirty Wistar rats (120–150g) were The volume of alkali was noted. This volume corresponds to
obtained from the animal house, Department of free acidity. Then 2 to 3 drops of phenolphthalein solution
Biochemistry, Karpagam Academy of Higher Education, were added and titration was continued until a definite red
Coimbatore. They were fed with grower mash (vital feed, tinge reappears. Again, the total volume of alkali added was
grand cereal) and water and kept for 2 weeks to acclimatize noted. This volume corresponds to total acidity. The free and
https://doi.org/10.25303/1804rjbt064073 65Research Journal of Biotechnology Vol. 18 (4) April (2023)
Res. J. Biotech.
total acidity of gastric juice was calculated by using the 95 percent ethanol and centrifuged at 3000 rpm for 30
following formula: minutes. The supernatant was removed in order to get a
glycoprotein precipitate which was utilized to calculate
Volume of NaOH x Normality of mucopolysaccharides such as sialic acid, hexose, fucose and
NaOH X 100 hexosamine.
Acidity =
meq/L
0.1
Assay of sialic acid: 0.1 N H₂SO₄ was used to hydrolyze
the glycoprotein precipitate of gastric juice. Following that,
Determination of ulcer index (UI) and percentage the hydrolysate was reduced with periodic acid and then
inhibition: Ulcer index and percentage inhibition were treated with thiobarbituric acid to liberate a color complex.
calculated in aqueous - induced rats for the initial studies of which was extracted with butanol and detected at 550nm25.
the efficacy of an optimized compound in an animal model.
Usually, circular lesions were observed but linear lesions Assay of total hexose: The gastric juice glycoprotein
were also seen. The ulcer index was calculated according to precipitate was treated with orcinol–H₂SO₄ reagent and
the scoring method36. heated at 80°C for 15 minutes to produce a color that was
detected at 540nm25.
Score 0 - no ulcer;
Score 1- vessel dilation and pointed ulcers; Assay of Fucose: The gastric juice glycoprotein precipitate
Score 2.5 - small ulcers 5 mm long. cystein hydrochloride. The difference in absorbance
measured at 393nm and 440nm was used as an indicator for
UI for each animal was calculated as mean ulcer score. The measuring the sample's fucose concentration25.
percentage of inhibition was calculated by the following
formula: Assay of hexosamine: HCl was used to dissolve the
glycoprotein precipitate. This hydrolysate was colored with
(UI control-UI treated) Erlich reagent and ethanol and measured at 530nm after
X 100 being treated with acetylacetone25.
UI control
Histopathology procedure: The different parts of the Assay of total non-amino polysaccharide: 0.1mL of
stomach were cleansed and fixed in 10% neutral buffered stomach juice was first precipitated with ethanol. After that,
formalin, dehydrated in graded series of alcohol, cleared in the precipitate was treated with bromosulfuric acid reagent
xylene and embedded in paraffin wax. The tissues were and tryptophan before being placed in a water bath to
sectioned at 5 µm with a rotary microtome and stained with produce color which was detected at 520nm25.
hematoxylin and eosin (H and E) and cresol violet stain11.
Estimation of lipid peroxidation: The technique of
Determination of free and total acidity: Volumetric Ohkawa et al27 was used to determine lipid peroxidation
analysis was used to measure the free and total acidity of the (LPO). 100 mL gastric juice and standard tubes containing
gastric juice, as described by Hawk15. To summarize, 0.5 mL MDA at concentrations ranging from 3 to 12 nM were
of gastric juice was pipetted into 100 mL conical flask, 2–3 combined with 0.2 mL sodium dodecyl sulfate, 1.5 mL
drops of Topfer's reagent were added and the mixture was acetic acid and 1.5 mL TBA. This combination was prepared
titrated with 0.01 N NaOH (previously standardized with up to 4mL with distilled water and maintained at 90°C for 1
0.01 N oxalic acid) until all traces of red color vanished and hour in a boiling water bath. 1mL distilled water was added
it turned into yellowish orange. Here hydrochloric acid after cooling to room temperature and the pink hue generated
(HCl) is typically not utilized in the titration process because was detected at 532nm against a reagent blank.
it reacts with the indicator potassium permanganate
(KMnO₄). It oxidizes after reacting with KMnO₄ solution, Estimation of protein: The protein content in the gastric
resulting in the release of chlorine gas. juice of pylorus-ligated rats was measured as described by
Lowry et al19.
The free acidity was calculated based on the volume of
titrated alkali. Then 2–3 drops of phenolphthalein solution Estimation of superoxide dismutase activity: Superoxide
were added and the titration was repeated until a clear red dismutase (SOD) (EC 1.15.1.1) activity in gastric juice was
tint emerged. The entire volume of alkali added was evaluated according to Marklund and Marklund21. 0.1 mL of
recorded once again. The overall acidity was represented by ethylenediamine tetraacetic acid (EDTA), 0.5 mL of
this volume. diethylenetriamine pentaacetic acid (DTPA) and 2.5 mL of
Tris-HCl buffer were mixed. To assess the rate of auto-
Estimation of glycoproteins: The glycoproteins found in oxidation of pyrogallol, 0.5mL of pyrogallol was added to
the gastric juice were isolated using Niebes25 modified the mixture and the increase in absorbance was detected at
technique1. 0.1 mL gastric juice was combined with 5 mL of 420nm for 3 minutes. 2.5mL of tris-HCl buffer, 0.1mL of
https://doi.org/10.25303/1804rjbt064073 66Research Journal of Biotechnology Vol. 18 (4) April (2023)
Res. J. Biotech.
EDTA and 0.5mL of DTPA were added to 0.1mL of gastric standard error of the mean (SEM) of the sample groups. Raw
juice obtained in a separate tube. 0.5mL pyrogallol was data were subjected to one way analyses of variance
added to this mixture and the rise in absorbance was (ANOVA) followed by post hoc Turkey‘s test.
measured at 420nm for 3 minutes. The rate of inhibition of
pyrogallol auto-oxidation caused by gastric juice was Results and Discussion
determined by this measurement. Peptic ulcer is one of the fastest growing disease in the
world. Despite many advances in the therapeutic
Estimation of catalase activity: The activity of catalase management of gastric ulcer, the prevalence of this disease
(CAT) (EC 1.11.1.6) in gastric juice was estimated by the is still high.
method of Sinha34. The optimum pH of catalase seems to be
pH 7.5. To 0.1mL of gastric juice, 1mL of phosphate buffer Phytochemical Analysis: Phytochemical test was carried
was added. Then 0.5mL of H₂O₂ was added to initiate the out on the aqueous crude extract as shown in table 1. The
reaction. The reaction was arrested at 0, 30 and 60 s intervals phytochemical screening of aqueous crude extract of
by the addition of 2mL of dichromate–acetic acid reagent. Sargassum wightii showed the presence of various chemical
The reagent blank was prepared by the addition of 1.6mL of constitutions mostly saponins, tannins, carbohydrates,
buffer and 2mL of dichromate–acetic acid reagent taken in flavanoids, alkaloids, glycosides, reducing sugar, proteins
separate tubes. The blank and the test tubes were heated in a which are conspicuously present in large amount.
boiling water bath for 10min to develop purple color. The
tubes were cooled to room temperature and their color These secondary metabolites have been reported to have
intensity was measured at 570nm against the blank. anti-ulcer activity34. Many phytochemical studies have
shown that plant compounds possess an important role in the
Statistical analysis: Data obtained from the study were prevention of gastric ulcer29. The composition of these
analyzed using Statistical Package for Social Sciences phytoconstituents depends upon the nature of the drugs and
(SPSS-21). The results were shown as the mean and the the solvent used.
Table 1
Phytochemical analysis of Sargassum wightii
S.N. Phytochemical Test Aqueous extract of Sargassum wightii
Alkaloids
1. a. Dragendorff’s test -
b. Mayer’s test -
2. Steriods -
3. Flavonoids
a. Magnesium turnings test +
b. Ferric chloride test +
4. Tannins
a. Ferric chloride test -
b. Gelatin test -
c. Lead acetate test +
5. Amino acids/ proteins
a. Ninhydrin reagent test +
b. Sodium hydroxide test -
6. Carbohydrates +
7. Cardioglycosides +
8. Saponin
a. Distilled water -
b. Vannilin test -
9. Terpenoids
a. Horizon test +
b. Libermann test -
+ = present, - = absent
Table 2
Acute toxicity study of the aqueous crude extract of Sargassum wightii18
S.N. Dose (mg/kg) Mortality
1. 150 mg/kg 0/3
2. 250 mg/kg 0/3
3. 500 mg/kg 0/3
https://doi.org/10.25303/1804rjbt064073 67Research Journal of Biotechnology Vol. 18 (4) April (2023)
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Acute Toxicity Studies (LD50): The acute toxicity test The crude extract Sargassum wightii showed a significant
carried out showed that the aqueous extract of Sargassum reduction (p < 0.05) in ulcer index, total acidity, gastric acid
wightii was highly safe having a high safety margin when volume and pH of gastric secretion when compared with the
compared to the standard range when given orally and this negative control distilled water (1 ml/kg) as shown in table
can be seen in the table 2. No deaths were recorded after 24 3.
hours of administration of the various doses (150, 250 and
500 mg/kg body weight) of the aqueous extract of The crude extract of Sargassum wightii also decreases the
Sargassum wightii. The LD50 was determined as severity and incidence of gastric erosions in Indomethacin
500mg/kg30. treated rat. For ulcer index, the rat treated with omeprazole
(3.00±0.52) and Sargassum wightii crude extract, at 150
Evaluation of anti - ulcer activity using Wistar rat model: mg/kg and 500mg/kg showed a significant reduction in ulcer
The antiulcer effect of the aqueous extract of the Sargassum index (7.10±0.20, 5.10±0.10, 3.65±0.1) than the rat
wightii showed the ulcer inhibition of the crude extract in pretreated with the negative control distilled water (1 ml/kg)
table 3. The antiulcer effect of the aqueous extract of the (12.05±0.20) at p < 0.05.
Sargassum wightii may be due to the flavonoids, saponins,
alkaloids and terpenoids which have been shown to produce For total acidity, the rat treated with omeprazole
some antiulcer and anti-gastric activity. The series of (57.3±0.3257.5±0.52) and Sargassum wightii ethanol
experiment were designed to determine the effects of extract, n-hexane, ethyl acetate, butanol and aqueous
Sargassum wightii aqueous extract on ulcer formation and fractions at 500mg/kg showed a significantly reduction in
its degree of toxicity. Indomethacin is a selective inhibitor of total acidity (57.3±0.32, 78.5±0.22, 85.0±0.00, 54.2±0.04)
cyclo-oxygenase and it strongly inhibits the cyclooxygenase than the rats treated with the negative control distilled water
pathway causing increased level of leukotriene in the gastric (1 ml/kg) (117.1±1.15) at p < 0.05.
mucosa which has been found to cause inflammation and
pain. It may also potentiate the secretory response elicited by For total acid volume, the rat treated with omeprazole
histamine due to the inhibition of prostaglandin E2. (6.46±0.14) at 500mg/kg showed a significant reduction in
total acidity (5.13±0.40, 7.02±0.004, 6.73±0.09, 5.82±0.21)
However, an anti-secretory effect might be indicated as the than the rats treated with the negative control distilled water
extract protected the stomach from NSAID (Indomethacin) (1 ml/kg) (6.46±0.10) at p < 0.05.
induced damage. This damage is elicited by the inhibition of
prostaglandin synthesis which is essential for mucosa Histology of the stomach of the Indomethacin - induced
integrity. This results to a sustained reduction mucosal blood ulcer in wistar rat: Histological examination of the stomach
flow and subsequent generation of ulcer28. of rat pretreated with negative control revealed several
changes in gastric mucosa such as severe desquamation and
Omeprazole is an effective agent in the treatment of peptic loss of surface epithelial cell, necrosis, vacuolization, edema
ulcer disease. Omeprazole is a proton pump inhibitor. Proton and dilated gastric glands along with infiltration of
pump inhibitors are the most effective anti-secretory agents inflammatory cells. The stomach of rat with the positive
available for the treatment of gastric acid-related disorders. control (Omeprazole) revealed few superficial surface of
mucosa: Gastric glands appeared normal without
These drugs inhibit basal and stimulated gastric acid inflammatory cells infiltration (Figure 1).
secretion by inhibiting the H+/K+-adenosine triphosphatase
(ATPase), also known as the proton pump which is located Histoarchitecture was not affected. The stomach of rat with
in the highly acidic luminal domain of the parietal cell. This aqueous extract of Sargassum wightii(500 mg/kg) showed
study has provided data suggesting that the aqueous crude decreased gastric lesions. Gastric mucosa exhibited focal
extract of Sargassum wightii contains biologically active loss of superficial gastric epithelium (Figure 2).
components that produce antiulcerogenic activities35.
Table 3
Dose dependent studies of aqueous crude extract of Sargassum wightii and Fractions
using indomethacin induced ulcer rat model.
Treatment Dose Ulcer Index Total Acidity (m Acid Volume pH (Control)
Eq/L) (ml)
Distilled water (1ml) 12.05±0.20 117.1±1.15 6.46±0.14 2.3±0.10
Omeprezole 20 mg/kg 3.00±0.52** 57.3±0.32** 5.13±0.40** 4.5±0.16**
Aqueous fraction 150mg/kg 7.10±0.20* 78.5±0.22* 7.02±0.04** 2.76±4.97**
Aqueous fraction 500mg/kg 3.65±0.1* 54.2±0.21* 5.82±0.04** 4.05±3.23**
Indomethacin 19.07±2.23** 114.3±1.15 6.53±0.14 2.6±0.14
Values are presented as mean ± Standard error of mean (SEM), * significant at p< 0.05, n=5.
https://doi.org/10.25303/1804rjbt064073 68Research Journal of Biotechnology Vol. 18 (4) April (2023)
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Control: C1-shows gastric mucosa with mucosal edema and moderate lymphocytic infiltration.
Mild ulceration occurs
C2-shows gastric mucosa with mucosal edema, mild lymphocytic infiltration.Mild ulceration occurs
Induced: I1-shows gastric mucosa with maintained glandular architecture and lamina propria showing mild
lymophytic infiltration. →No evidence of ulceration
I2-shows gastric mucosa with edema, areas of haemorrhage and infiltration. →No evidence of ulcerat
Figure 1: Histopathology of stomach tissue of control and induced rat
Standard: S1-shows gastric mucosa with maintained glandular architecture and lamina propria showing mild
lymophytic infiltration. →No evidence of ulceration
S2-shows gastric mucosa with mucosal edema and moderate lymphocytic infiltration. No evidence of ulceration
Extract: E1-shows gastric mucosa with maintained glandular architecture and lamina propria showing mild
lymophytic infiltration. →No evidence of ulceration
E2-shows gastric mucosa with mild lymophytic infiltration. →No evidence of ulceration
Figure 2: Histopathology of stomach tissue of standard and extract rat
The gastric glands were almost normal in appearance. The setting was based on visual endoscopy, but this study was
healing process of gastric ulcer included several processes in based on microscopic evaluation and gastric ulcer
gastric mucous e.g. congestive, hemorrhagic, edema, determination. Microscopic evaluation showed gastric
necrosis, inflammation, erosion, ulceration and dysplastic glands dilatation, increase of connective tissue, increase of
change. Evaluation for the healing process in the clinical micro vascularization and recovery of a sensory nerve. It
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could be the basis for evaluating the quality of the healing activity of ethanol leaf extract of Cassia fistula observed in
process of gastric ulcers26. this study could be due to the protection of the mucosal
barrier system of gastric mucosa in pylorus-ligated rats5.
Effect of crude sea weed on glycoproteins in the gastric
juice: The effect of Sargassum wightii of indomethacin- Effect of Lipid peroxidation on aqueous extract of
treatments on the status of glycoproteins in the gastric juice Seaweed treatment: Omeprazole pre-treatment (group II)
is presented in table 4. The data show a fall in the status of significantly decreased the levels of lipid peroxidation in the
sialic acid and fucose in omeprazole-treated rat (group II) as gastric juice of rat when this group was compared to positive
compared to the positive control (group I). In contrast, a control (group I). Pre-treatment with aqueous extract of
highly significant increase in the status of total hexose, seaweed produced a dose-dependent decrease in
hexosamine, total non-amino polysaccharide (TNAP) and peroxidation in the omeprazole -ligated rat (groups III) and
total carbohydrate was observed in group II rats when at the highest dose (group IV), the decrease in the above
compared to group I. Treatment with crude aqueous extract parameters was comparable to omeprazole treated animals
Sargassum wightii at 150 and 500 mg/kg (groups III and IV (Figure 3).
respectively) also caused a highly significant decrease in
sialic acid and fucose accompanied by an increase in all the Effect of catalase on aqueous extract of Sea weed
other glycoproteins in a dose-dependent manner, when these treatment: Omeprazole pre-treatment (group II)
groups were compared to control. significantly decreased the levels of catalase in the gastric
juice of rat when this group was compared to positive control
Treatment of rat at a higher dose of crude aqueous extract (group I). Pre-treatment with aqueous extract of seaweed
Sargassum wightii (group IV) produced an effect produced a dose-dependent decrease in catalase in the
comparable to that in omeprazole-treated rat in the status of omeprazole-ligated rat (groups III and IV) and at the highest
all the glycoproteins investigated in the gastric juice of dose (group III) the decrease in the above parameters was
indomethacin - rat. It was already reported that the antiulcer comparable to omeprazole treated animals (Figure 4).
Table 4
Levels of glycoproteins in gastric juice of Sargassum wightii
Determinant Control Standard Extract Extract Induced
(µg/mL) (150mg/kg) (500mg/kg)
Sialic acid 45± 0.08 51± 0.08 74± 0.05 69± 0.03 35± 0.06
Hexosamine 49± 0.07 54± 0.05 73± 0.06 67± 0.05 40± 0.07
Total hexose 50± 0.05 53± 0.06 74± 0.04 66± 0.06 41± 0.05
TNAP 47± 0.04 50± 0.03 67± 0.08 63± 0.04 37±0.01
Fucose 50± 0.04 54± 0.03 68± 0.07 62± 0.06 36± 0.04
Effect of Lipid Peroxidation
0.7
0.6
nmoles/min/mg Protein
0.5
0.4
0.3
0.2
0.1
0
Control Standard Extract Extract Induced
(150mg/kg) (500mg/kg)
Figure 3: Effect of aqueous extract of Seaweed on lipid peroxidation (LPO) in the gastric juice of indomethacin
induced rat. Group I was treated with saline (Control). Group II was treated with 20mg/kg of Omeprazole
(standard). Groups III and IV were treated with aqueous extract of Seaweed at 150 and 500mg/kg respectively
(Extract). Group V was induced with Indometacin. Results are given as mean ± SEM of six numbers
of animals in each group.
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Figure 4: Effect of aqueous extract of Seaweed on Catalase (CAT) in the gastric juice of indomethacin induced rat.
Group I was treated with saline (Control). Group II was treated with 20mg/kg of Omeprazole (standard).
Groups III and IV were treated with aqueous extract of Seaweed at 150 and 500mg/kg respectively (Extract).
Group V was induced with Indometacin
Effect of superoxide dismutase
0.5
0.45
0.4
0.35
Units/mg protein
0.3
0.25
0.2
0.15
0.1
0.05
0
Control Standard Extract Extract Induced
(150mg/kg) (500mg/kg)
Figure 5: Effect of aqueous extract of Seaweed on Superoxide dismustase in the gastric juice of indomethacin induced
rat. Group I was treated with saline (Control). Group II was treated with 20mg/kg of Omeprazole (standard).
Groups III and IV were treated with aqueous extract of Seaweed at 150 and 500mg/kg respectively (Extract).
Group V was induced with Indometacin
Effect of superoxide dismutase on aqueous extract of Effect of protein on aqueous extract of Seaweed
Seaweed treatment: Omeprazole pre-treatment (group II) treatment: While the total protein was decreased
significantly decreased the levels of SOD in the gastric juice significantly by about 40%, the protein content was elevated
of rat when this group was compared to positive control by about two-fold in the gastric juice of omeprazole pre-
(group I). treated rat (group II) when they were compared to control
(group I). Although pre-treatment with aqueous extract of
Pre-treatment with aqueous extract of seaweed produced a seaweeds at 100 and 500mg/kg (groups III and IV) also
dose-dependent decrease in SOD in the omeprazole-ligated produced an effect similar to that in omeprazole –treated rat,
rat (groups III and IV) and at the highest dose (group III) the their efficacy was not comparable to this group. On the other
decrease in the above parameters was comparable to hand, pre-treatment with indomethacin (group V) produced
omeprazole treated animals (Figures 5). an effect comparable to control rat (Figure 6).
https://doi.org/10.25303/1804rjbt064073 71Research Journal of Biotechnology Vol. 18 (4) April (2023)
Res. J. Biotech.
Effect of Protein
0.8
0.7
Units/mg Protein 0.6
0.5
0.4
0.3
0.2
0.1
0
Control Standard Extract Extract Induced
(150mg/kg) (500mg/kg)
Axis Title
Figure 6: Effect of aqueous extract of Seaweed on protein in the gastric juice of indomethacin induced rat.
Group I was treated with saline (Control). Group II was treated with 20mg/kg of Omeprazole (standard).
Groups III and IV were treated with aqueous extract of Seaweed at 150 and 500mg/kg respectively (Extract).
Group V was induced with Indometacin.
Conclusion B.L., Antiulcerogenic activity of Scutia buxifolia on gastric ulcers
Pharmacognostic properties help in the identification and induced by ethanol in rats, Acta Pharm Sin B, 4, 358-67 (2014)
authentication as a reliable tool for the standardization and
5. Bruce S.O. and Okoye C., Pharmacognostic, phytochemical and
quality evaluation of the seaweed. This study indicates that antiulcer properties of ethanol crude extract and fractions of the
the Sargassum wightii is a safe seaweed. The anti- leaves of picralima nitida durand and hook (apocynaceae), World
ulcerogenic properties exhibited that at 500mg/kg it showed Journal of Pharmaceutical Research, 11(1), 20-40 (2022)
a good anti-ulcer activity as compared with the standard drug
20mg/kg omeprazole statistically. Thus, the present work 6. Coura C.O., DeAraujo I.W., Vanderlei E.S., Rodriques J.A.,
validates the use of Sargassum wightii for gastric ulcer. Quindere A.L., Fontes B.P., de Queiroz I.N., de Menezes D.B.,
Bezerra M.M., E Silva A.A., Chaves H.V., Jorge R.J., Evangelista
Acknowledgement J.S. and Benevides N.M., Antinociceptive and anti-inflammatory
activities of sulphated polysaccharides from the red seaweed
The authors are thankful for the financial support granted by Gracilaria cornea, Basic Clin Pharmacol Toxicol, 110, 335-341
KAHE under Seed Money Project No. (KAHE/R- (2012)
Acad/A1/Seed Money/036). The first author is grateful to
the authorities of Karpagam Academy of Higher Education 7. Da Silva L.M., Allemand A., Mendes D.A.G., dos Santos A.C.,
for the financial assistance for herbal project. All the authors André E., de Souza L.M., Cipriani T.R., Dartora N., Marques
are grateful to the authorities of Karpagam Academy for M.C.A. and Baggio C.H., Ethanolic extract of roots from Arctium
providing the necessary facilities to carry out this work. Dr. lappa L. accelerates the healing of acetic acid-induced gastric ulcer
M. Ganesan is acknowledged for collecting and identifying in rats: Involvement of the antioxidant system, Food Chem.
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