Antiulcer potential of Chloroform extract of Cucumis sativus 2018

7.0 RESULTS

7.1 Phytochemical evaluation:

Table 9: Organoleptic characters of C. sativus

Organoleptic characters C. sativus
Colour Greento brown
Odour Characteristic
Surface characteristics Glossy and leathery
Shape Cylindrical and tapered

Table 10: Physiochemical parameters of powdered fruits of C. sativus

S. no. Parameters Observed value (%w/w)
1 Foreign matter Nil
2 Water soluble extractive value 6.8
3 Alchoholic soluble extractive value 2.3
4 Total ash value 6.1
5 Water soluble ash value 3.1
6 Acid insoluble ash value 1.3
7 Sulfated ash value 0.9
8 Loss on drying /Moisture content 3.2

Result and discussion

It was found that there were no visible signs of plant, animal and mineral foreign matter.It
indicates that powder of C. sativus found to be free from foreign matters contamination. The
water soluble extractive value of C. Sativus was found to be 6.8 % w/w. It indicates that C.
sativus plant contain water soluble Phytoconstituents.

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
The alcohol soluble extractive value of C. Sativus was found to be 2.3 % w/w. Alcohol soluble
extractive value is used to determine the alcohol soluble Phytoconstituents of crude plant drug. It
indicates that C. sativus plant contain ethanol soluble Phytoconstituents. The total ash value of C.
sativus was found to be 6.1 % w/w. The total ash method used to measure the total amount of
material remaining after ignition. This includes both physiological and non-physiological ash
which is the residue of the extraneous matter adhering to the plant surface.

The water soluble ash value of C. Sativus was found to be 3.1 % w/w. The acid-insoluble ash
value of C. Sativus was found to be 1.3 % w/w. Acid insoluble ash value is used to determine the
percentage of siliceous matter like silicate and sand. The sulphated ash value of C. Sativus was
found to be 0.9% w/w.

Moisture content of C. Sativus was found to be 3.1 % indicates that the plant contain moisture
within the limit (not more than 5 %). Moisture content determination related to the stability of
powdered drug because excess of moisture content cause breakdown of Phytoconstituents by
enzymatic activity and may encouraged the growth of yeast and fungi during storage. As the
height of foam in every test tube was found to be less than 1 cm so the foaming index is less than
100which indicates that C. sativus plant contain high saponins contents.

Table 11: Percentage Yield of Various Extracts of C. sativus

S. No. Solvent used for Timebrequired for Colour of Percentage of

Extraction Complete extraction Extract yield(w/w)
(Hrs.)
1. Petroleum ether 40 Darkgreen 6.35%

2. Chloroform 40 Darkgreenish 7.26%

Brown

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
Results and discussion
The percentage yield of petroleum ether, chloroform and extract of C. sativus fruits was found to
be 6.35 %, 7.26 % respectively.These extracts and fractions were stored in airtight container for
further studies.

Table 12: Qualitative chemical tests of different extracts of C. sativus fruits

Petroleum ether Chloroform
Extract Extract
Tests
Steroid + -
Triterponoids - -
Glycosides - +
Carbohydrates - -
Alkaloids - +
Flavonoids - +
Tannins - -
Proteins and amino acid - -
Lipids + -

Results and discussion

Petroleum ether extract fruits showed presence of steroid, chloroform extract showed the
presence of glycoside, alkaloid, flavanoid.

Table1 3 : T.L.C plate of C. sativus extracts under visible light

S.no. Phytoconstituents Numbers of spot Rf Value

1 Flavonoids 1 0.75
2 Glycoside 1 0.48
3 Tannin 2 0.76
4 Alkaloid 2 0.11, 0.43

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
Results and discussion
The presence of major chemical constituents from various extracts was confirmed by TLC
studies. TLC of various extract of C. sativus was showed the presence of flavonoids, glycosides,
tannins and alkaloids.

7.2 Pharmacological evaluation:

(A) Dose Dependent Studies:

Table No.14: Dose Dependent Studies

S. No. Drug Dose Route of No. of Parameters for

Administration Animal Study

Control --------  Ulcer

1. (water) Oral 6 index

Standard 3mg/kg  Total

2. (famotidine) Oral 6 acidity

Fresh extract of C. 0.5ml/100gm  Acid

3. Sativus Oral 6 volume

Fresh extract of C. 1.0ml/100gm  pH

4. Sativus Oral 6  Glutathi

one
Fresh juice of C. 2.0ml/100gm
 Total
5. Sativus Oral 6
protein

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
(B) Time Dependent Studies :
Table No.15: Time Dependent Studies
S. Drug Dose No. of Durations Parameters for study

No. Animals

1. Control -----------  Ulcer index

(water) 6 15 Days

2. Standard 3mg/kg  Total acidity

(famotidine) 6 15 Days

3. Fresh extract 1.0ml/100gm

of leaves of 6 15 Days  Acid volume

C. sativus

4. Control -----------  pH

(water) 6 30 days

5. Standard 3mg/kg
 Glutathione
(famotidine) 6 30 days

6. Fresh extract 1.0ml/100gm

 Total protein
of C. sativus 6 30 days

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 Antiulcer potential of Chloroform extract of Cucumis sativus 2018
(C) Dose Dependent Studies:

Table No.16: Dose dependent studies of the fresh extract of fruits of C. sativus using rats in
ethanol induced ulcer model.

S. Treatment No of Dose Ulcer Index Total Acid pH

N. Animal Acidity Volume
s (mEq/L) (ml)

1 Control 6 --- 10.14±0.32 117.1±1.12 8.31±0.24 2.3±0.12

(water)

2 Famotidine 6 0.3ml/100gm 4.23±0.32** 55.6±0.54** 4.65±0.31** 4.94±0.12**

3 FEFCS 6 0.5ml/100gm 8.76±0.21 95.5±0.22* 6.55±0.13* 3.22±0.27

4 FEFCS 6 1.0ml/100gm 6.19±0.31* 75.0±0.56* 5.23±0.19* 3.81±1.17**

5 FEFCS 6 2.0ml/100gm 1.45±0.21** 62.3±0.31** 5.03±0.07** 4.13±0.53**

FJLPM - fresh juice of leaves of C. sativus

**P<0.001, *P<0.05, compared with control.

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018

Dose dependent studies of the fresh extract of fruits of C. sativus using

rats in ethanol induced ulcer model.

12
10
8
6
4 Ulcer Index
2 Acid
0 pH

Figure 4 -Dose dependent studies of the fresh extract of fruits of C. sativus using rats in
ethanol induced ulcer model.

Total Acidity
120
100
80
60
40
20
Total Acidity
0

Figure 5-Total Acidity

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 Antiulcer potential of Chloroform extract of Cucumis sativus 2018
(D) Time Dependent Studies
Table No.:17 Time dependent studies of the fresh extract of fruits of C. sativus using rats in
ethanol induced ulcer model.

S. Treatment No Dose Treat Ulcer Total Acid Ph

No of ment Index Acidity Volume
Ani Durati (mEq/L) (ml)
mal on
s (days)
1 Control 6 --- 15 12.4±0.32 118.8±3.43 8.35±0.42 2.52±0.16

2 Famotidine 6 0.3ml/100gm 15 4.3±0.41 59.7±0.91** 4.59±0.25** 5.13±0.12**

3 FEFCS 6 1.0ml/100gm 15 1.28±0.17 67±0.35* 4.29±0.08 4.31±0.05#

4 Control 6 --- 30 11.6±0.30 116±0.44** 7.95±0.16 2.7±0.08

5 Famotidine 6 0.3ml/100gm 30 0.17±0.03* 58.5±0.22** 3.75±0.12 5.0±0.06**

6 FEFCS 6 1.0ml/100gm 30 0.43±0.01* 55.5±0.22 4.05±0.02* 5.2±0.02**

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018

Time dependent studies of the fresh extract of fruits of C. sativus using rats
in ethanol induced ulcer model.

14
12
10
8
6
4 Ulcer Index
2
Acid Volume (ml)
0
pH

Figure 6-Time dependent studies of the fresh extract of fruits of C. sativus using rats in ethanol
induced ulcer model.

Total acidity (mEq/L)

120
100
80
60
40
20 Total acidity (mEq/L)
0

Figure No. 7 -Total acidity (mEq/L)

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
Table No.:18 Time dependent studies of the fresh extract of fruits of C. sativus using rats in
ethanol induced ulcer model

S.No Treatment No of Dose Treatment Glutathione Total Protein

animals Duration (mcg/gm) gm/Dl

(days)

1 Control 6 - 15 0.81±0.11** 0.836±0.18

2 Famotidine 6 0.3ml/100gm 15 1.61±0.36 0.620±0.16**

3 FEFCS 6 1.0ml/100gm 15 1.13±0.13* 0.541±0.13

4 Control 6 - 30 0.84±0.11 0.812±0.11

5 Famotidine 6 0.3ml/100gm 30 1.72±0.12** 0.538±0.21*

6 FEFCS 6 1.0ml/100gm 30 1.18±0.07 0.495±0.s12

FJLPM - fresh juice of leaves of C.sativus

**P<0.001, *P<0.05, compared with control.

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
Time dependent studies of the fresh extract of fruits of C. sativus using rats in
ethanol induced ulcer model.

1.8
1.6
1.4
1.2
1
0.8
0.6
0.4 Glutathione (mcg/gm)
0.2
0 Total Protein gm/dL

Figure 8-Time dependent studies of the fresh extract of fruits of C. sativus using rats in ethanol
induced ulcer model.

Result and discussion:

Peptic ulcer is defined as disruption of the mucosal integrity of the stomach and/ or duodenum
leading to a local defect or excavation due to active inflammation. Despite the constant attack on
the gastroduodenal mucosa by a host of noxious agents (acid, pepsin, bile acids, pancreatic
enzymes, drugs, and bacteria), integrity is maintained by an intricate system that provides
mucosal defense and repair. This intricate biologic system consist of mucusbicarbonate layer,
surface epithelial cells and a rich submucosal micro-circulatory bed which provides bicarbonate
ions to neutralize the acid generated by parietal cell secretion of hydrochloric acid. Moreover,
this microcirculatory bed provides an adequate supply of micronutrients and oxygen while
removing toxic metabolic by products. Various studies suggested that changes in gastric motility
may play a role in the development and prevention of experimental gastric lesions.

Relaxation of circular muscle may protect the gastric mucosa through flattening of the folds.
This will increase the mucosal surface area exposed to necrotizing agents and reduce the volume

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
of the irritant on the rugal crests. Such an action has been postulated to play a role in the
cytoprotective effect of prostaglandins.

The present study has been done to evaluate the antiulcer effect of fresh juice of the leaves of C.
sativus on ethanol induced time and dose dependent ulcers. The results obtained from the present
study have been shown that fresh juice of the leaves of C. sativus possesses antiulcer effect on
ethanol induced ulcers. In ethanol and stress induced model, there is decrease in ulcer index, total
acidity, total volume of gastric secretion, total protein and increase in glutathione content and pH
of gastric secretion when compared with control. In the present study Famotidine has been used
as a standard.

Ethanol induced gastric ulcer:

Ethanol serves as a most common ulcerogenic agent and when given intragastrically to rats it
produces severe gastric hemorrhagic erosions. The genesis of ethanol induced gastric lesions is
multifactorial with the depletion of gastric wall mucus content as one of the involved factors and
this damage induced by ethanol may be due to mucosal leukotriene release. Mucosal blood flow
has also been attributed to be an important factor in the damage caused by alcohol and is
modulated by prostaglandin. Submucosal venular constriction by ethanol and eventual injury is
caused due to perturbations of superficial mucosal cells, notably the mucosal mast cells leading
to release of vasoactive mediators including histamine, that cause
damage to gastric mucosa. Ethanol-induced damage to the gastric mucosa is associated with a
significant production of free redicals leading to an increased lipid peroxidation and damage to
the cell and cell membranes. Accumulation of activated neutrophils in the gastric mucosa may be
a source of free radicals.

In the present study, Famotidine used as a standard drug. Famotidine is a H2 receptor blocker, is
capable of reducing over 90% of basal, food stimulated and nocturnal secretion of gastric acid,
stimulated by histamine, gastrin, cholinomimetic drugs and vagal stimulation. Famotidine exerts
its antisecretory effect by inhibiting the histamine induced c-AMP dependent pathway.
Famotidine also increase certain mucus component of gastric mucus in patients with duodenal
ulcer.

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018

Total acidity indicates that how much acid is present in the gastric secretion. It is a major
aggressive factor which produce ulcer. Gastric secretion is under vagal control and over activity
of vagus also contributes to ulcer formation. Vagal stimulation releases acetylcholine that act
directly on muscarinic receptors on parietal cells and secretes hydrochloric acid through a Ca++
dependent pathway.

On ethanol treatment, the mucosal mast cells leading to release of vaso active mediators
including histamine. Histamine is thought to stimulate the production of cyclic AMP by
activating the enzyme adenylcyclase which causes activation of gastric proton pump and release
of hydrogen ions. FEFCS treatment showed decrease in the total acidity of the gastric secretion.
Serum protein consists of albumin and globulin. In the peptic ulcer the total protein content of
serum or gastric juice is increased. This could be due to leakage of plasma protein in to the
gastric juice or serum with weakening of the mucosal resistance/barrier of the gastric mucosa.

After treatment with FEFCS there was a significant decrease in protein content of gastric juice
which suggest that it might be primarily decrease the leakage of plasma protein in to the gastric
juice with strengthening of the mucosal barrier and increase in its resistance to the damaging
effect of aggressive factors.

Acid volume is a volume (ml) of acid present in the gastric secretion which contains HCl,
pepsinogen, mucus, bicarbonates, intrinsic factor and proteins. Volume of acid secretion is also
an important factor in the production of ulcer due to exposure of unprotected lumen of the
stomach to the accumulating acid. FEFCS treatment showed decrease in the acid volume of the
gastric secretion.

Higher pH indicates a lower concentration of the hydrogen ion. The hydrogen ion is a major
aggressive factor involved in the genesis of ulcer and gastric damage. FEFCS treatment showed
a increase in pH of the gastric contents. This directly indicates that the FEFCS lowers the
chances of ulcer and has a protective action upon the gastric mucosa.

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Antiulcer potential of Chloroform extract of Cucumis sativus 2018
In gastric ulcer tissues, Glutathione (g-glutamylcysteinylglycine, GSH) levels were found to be
decreased. Ethanol-induced generation of free radicals reduces the cysteine which is required for
GSH synthesis, which is, there for, decreased. Data from this study indicated that depletion of
gastric GSH is associated with generation of gastric lesion in the rats. GSH is a tripeptide and a
superoxide radical scavenger and it protects thiol protein groups required for maintaining the
integrity of cell against oxidation. In our present study, FEFCS treatment showed increase in the
glutathione content.

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