Evaluation of Hydro-alcoholic extract of Corchorus capsularis seeds for its Anti-ulcer activity on Albino Wistar Rat

 

Kousik Saha¹*, Trisha Sen¹, Mrityunjoy Majumdar¹, Hira Das¹, Oindrila Baisya²

¹Department of Pharmacology, Netaji Subhas Chandra Bose Institute of Pharmacy,

Chakdaha, Nadia 741222, West Bengal, India.

²Department of Pharmaceutics, Netaji Subhas Chandra Bose Institute of Pharmacy,

Chakdaha, Nadia 741222, West Bengal, India.

 

ABSTRACT:

The objective of present study was to evaluate the anti-ulcer activity of hydro-alcoholic extract of Corchorus capsularis seeds. Mainly hyper secretion of gastric acid and pepsin causes ulcer in patients. Plant extracts and polyherbal formulations are two of the most appealing sources of new drug. This type of extract formulations are showing good results in the treatment of gastric ulcers. In this study anti-ulcer activity was investigated by pyloric ligation, aspirin and stress induced gastric ulcer in albino wistar rats. The extract was given at the dose of 250mg/kg (CCLD- Corchorus capsularis low dose) and 500mg/kg (CCHD- Corchorus capsularis high dose) in all the experimental models. Ranitidine (150mg/kg) was used as standard drug. CCLD and CCHD showed 37.49% and 63.14% ulcer inhibition respectively, whereas ranitidine 69.69% against pylorus ligation induced ulcer. In aspirin induced ulcer model the extract showed ulcer inhibition 48.64% and 64.00% in CCLD and CCHD respectively whereas the standard drug ranitidine exhibited 84.64% ulcer inhibition. In stress induced ulcer model CCLD, CCHD and ranitidine exhibited 48.64%, 64.00% and 84.64% ulcer inhibition respectively. In all the models ethanolic extract of Corchorus capsularis seeds has shown significant anti-ulcer activity in dose dependent manner.

 

 

INTRODUCTION:

Currently, ulceration is a common problem around the world and remains an important cause of increased morbidity and health care costs. Ulcer is a gastrointestinal disease, affects 10% of the world population¹. According to the WHO reports published in 2018, the total deaths in India have reached up to 55,560 or 0.63% due to ulcer and India ranks 60 in the world with the age-adjusted death rate is 5.58 per 100,000².

 

Ulcer is defined as a change in the integrity of mucosal barrier of the stomach and/or duodenum, causing local defect or excavation due to active inflammation³.

 

Ulcer occurs in stomach is known as gastric ulcer and in the first part of small intestine it is called duodenum ulcer. It is one of the major gastrointestinal (GI) disorders, which occurs because of imbalance in the offensive factors such as gastric acid, pepsin, bile, H. pylori and defensive factors such as gastric mucosal integrity, prostaglandins, bicarbonate secretion and innate resistance of the mucosal cells⁴. Common causes of ulcer are Helicobacter pylori, chronic usage of NSAIDs, smoking, alcohol intake, excessive stress etc. Upper abdominal pain, belching, and vomiting with some complications such as bleeding, perforation, and blockage of stomach are the symptoms of ulcer. Basically, treatment of ulcer involves symptoms reduction, ulcer healing and avoiding reappearance. The medications used to treat ulcer are either decrease acid secretion i.e. proton-pump inhibitor, H₂ blocker, and prostaglandin analogues or neutralize stomach secreted acid i.e. antacids. However, the majority of these drugs produce undesirable side effects. Due to numerous side effects associated with synthetic drugs, medicinal plants are considered as a primary source of new drugs as they have fewer or no side effects and they are also economical, effective and less toxic⁵.

 

The plant Corchorus capsularis commonly known as jute belongs to the family Malvaceae. It is mainly cultivated in India, Bangladesh, Bhutan and some other countries in the world. Usually the plants are 2-4 m tall annual herb. The leaves are simple, alternate and 5-15 cm long with acuminate tip. The flowers have five petals, small and yellow colour. Capsules are small, oval shaped and wrinkled⁶. The seeds contain cardiac glycosides like corchorin, corchortoxin, helveticoside, corchoroside A and B⁷. It also contains biosides, erysimoside, oligosaccharide, raffinose, stearic acid and some other micro and macro elements⁸.

 

Corchorus capsularis is used in the folk remedy for aches, pains, dysentery, enteritis, fever, tumours and also used as a gastro-protective agent. Various studies showed that it has cardiotonic⁹, anticancer¹⁰, antioxidant¹¹, anti-inflammatory, analgesic and antipyretic¹², antimicrobial¹³ and insecticidal¹⁴ activity. Present study was designed to scientifically evaluate the antiulcer activity of Corchorus capsularis seeds extract at different doses.

 

MATERIALS AND METHODS:

Experimental Animals:

Female albino wistar rats of 150-200g were used throughout the experiment. They were placed in polypropylene cages (32×24×16cm). The animals were purchased from authorised animal breeder. The animals were kept in CPCSE approved Netaji Subhas Chandra Bose Institute of Pharmacy animal house (approval no:1502/PO/a/11/CPCSEA), well maintained under standard hygienic conditions, at a temperature of 22±2°C, 65% relative humidity, and 12-hours light and dark cycle. Commercial food pallets and tap water ad libitum were provided. All the experiments were performed in between 10am to 6pm.

 

Drugs and chemicals used:

Ethanol (Loba Chemie Pvt. Ltd.), sodium hydroxide (Loba Chemie Pvt. Ltd.), hydrochloric acid (Loba Chemie Pvt. Ltd.), diethyl ether (Merck Specialities Pvt. Ltd., Mumbai.), ranitidine (Aciloc 150manufactured by Cadila Pharmaceuticals Ltd.), aspirin, distilled water.

 

Preparation of extract formulation:

The seeds of Corchorus capsularis were purchased from local market. They were washed by clean and fresh water to remove the unwanted particles. The seeds were dried, powdered and then extracted with 70% ethanol in soxhlet apparatus for about 48 hours. Then the extract was dried by using a water bath and kept at 4⁰C in refrigerator until the experimental testing.

 

Preliminary phytochemical investigations of the extract:

Hydro-alcoholic extract of Corchorus capsularis seeds was evaluated for phytochemical investigation¹⁵ and presence of phytoconstituents like alkaloid, carbohydrate, saponin, flavonoid, glycoside, sterol, protein etc. were observed.

 

Experimental design:

For this study the animals were divided into four groups of six animals in each group.

·       Group I

The animals of this group was untreated and known as control.

·       Group II

This group received the treatment of ranitidine 150 mg/kg orally.

·       Group III

This group was treated with 250 mg/kg dose of Corchorus capsularis seeds extract orally. This group was known as CCLD.

·       Group IV

      This group was treated with 500 mg/kg dose of Corchorus capsularis seeds extract orally and known as CCHD.

 

Pyloric ligation induced ulcer^16-19

In this method, all the animals were kept for 24 hours fasting before initiation of the experiment and allowed free access to drinking water. All the animals were given treatment as per their groups 30 minutes before the ligation. Then the animals were anaesthetised by diethyl ether. After ligation the sutures were given in the operated area. Water supply was restricted on postoperative period. After 4 hours the animals were sacrificed, the stomachs were isolated and opened. Stomach contents were collected into tubes and total volume of gastric content was measured. Centrifugation of gastric content was done at 1000 rpm for 10 min. About 1 ml of the supernatant liquid was pipette out and diluted with distilled water up to 10 ml. This solution was titrated against 0.01 N NaOH using Topfer’s reagent as an indicator.  NaOH volume was measured and acidity was calculated as on:

 

                Voloume of NaOH × Normality

Acidity = ––––––––––––––––––––––––––– × 100

                                mEq/L/100 g

 

Formation of any lesion in the stomach wall was observed and examined, the number of ulcers were recorded and scored as:

0 = No ulcer ; 1 = Superficial ulcers; 2 = Deep ulcers; 3 = Perforation.

 

Mean ulcer score for each animal was expressed as ulcer index (U_I):

U_I    = U_N + U_S+ U_P × 10^–1

U_N = Mean value of ulcers/animal

U_S = Average of severity score

U_P = Percentage of animals with ulcers

 

The acidity and ulcer index of the treated animals were compared with controls. The ulcer inhibition (%) was calculated as: 

 

                       Mean ulcer index of control - Mean ulcer index of test

Inhibition (%) = –––––––––––––––––––––––––––––––––––––– × 100

                                               Mean ulcer index of control

 

Aspirin induced ulcer^20-22:

In this model ulcer was induced by aspirin administration. Prior to the start of the experiment, all the rats were kept for 24 hours fasting and given free access to drinking water. The animals were given treatment as per their groups, after 45 minutes of the treatment, all rats were received aspirin (200mg/kg, orally) to induce gastric ulcer. After 4 hours the animals were sacrificed by inhalation of anaesthetic ether. The stomachs were isolated, dissected and cleaned to remove gastric contents and blood by using distilled water. The number of ulcers and the severity scores were recorded. The acidity, ulcer index and percentage of ulcer inhibition was calculated by the formulas described in pyloric ligation model.

 

Stress induced ulcer by water immersion^23-26:

In this model ulcer was induced by immersing the animals in water for 4 hours. All the animals were kept in 24 hours fasting before initiation of the experiment and given free access to drinking water. The animals were given treatment as per their groups, after 30 minutes of the treatment, the animals were placed in a cage and immured in a water tank (20°C–23°C) for 4 hours. After 4 hours, the animals were taken out from the water immersion cage and sacrificed. The stomach of each animal was isolated, mounted and the number of ulcer formation was counted for evaluation. The severity scores were noted. The acidity, ulcer index and percentage of ulcer inhibition was calculated by the formulas described in pyloric ligation model.

 

Statistical analysis:

The experimental result were shown as mean±SEM for each treatment group. The significance of activity was assessed using one-way ANOVA, Dunnett’s post-parametric test between the data of control and treated groups. *p<0.001 was considered statistically significant.

RESULTS AND DISCUSSION:

Gastric ulcer occurs because of imbalance in the aggressive and defensive factors. The basolateral membrane of parietal cell has four important receptor namely muscarinic (M₁), histaminic (H₂), gastrin (G) and prostaglandin (P), among them M₁, H₂ and G are responsible for positive action on proton pump, may enhance acid secretion in stomach²⁷. Prostaglandin produces inhibitory signal on proton pump, reduces the acid secretion. On the other hand it enhances the mucus production which produces a protective layer in stomach.

 

Ulcer is a burning issue mostly in the Eastern India, may be due to incorporation of more spices in the dishes of this region. Lots of drugs from different medicinal system like allopathic, homeopathic, ayurveda and unani are available to combat this situation. As per, depending upon the seriousness of the situation may many combination drug therapy are also used in India, but the most popular antiulcer drugs from allopathic system has major draw-backs like side effects and high cost. Some of the herbal drugs has a certain level of anti-ulcer activity but there are always scopes for exploration of new molecules which can be easily available and cost-effective.

 

In this study Corchorus capsularis seeds have been investigated for its antiulcer activity by using three different model namely pyloric ligation induced ulcer, aspirin induced ulcer and stress induced ulcer. In all the model ranitidine (H₂ antihistaminic) has been taken as taken as standard drug. The selection of the drug as a standard has come to the idea due to its no anti-androgenic and CNS effect.

 

In the first model namely pyloric ligation induced ulcer in rat the pylorus has been ligated with the intention to accumulate the secreted acid in the stomach within a stipulated period of time. The accumulated acid has shown a tendency towards ulcer formation in stomach. In the control group, where the animals had not received any treatment, all the animals were affected with number of ulcerative lesion in stomach. Pretreatment with ranitidine has given major protection towards the ulcer formation in rat. Corchorus capsularis seeds extract in two different doses i.e. CCLD and CCHD has shown different level of protection. In two different parameters i.e. ulcer index and total acidity it was clear that ranitidine and Corchorus capsularis extracts have shown significant decrease in ulcer index and total acidity while compared with untreated control. In the present study ranitidine, CCLD and CCHD have shown 69.69 %, 37.49 % and 63.14 % ulcer inhibition (Table 1) respectively when compared with untreated control.

 

 

Figure 1: Sample of dissected stomach and formation of ulcer in pyloric ligation induced ulcer model. (a) Control, (b) ranitidine, (c) CCLD, (d) CCHD.

All values are shown as mean ± SEM, n = 6. ***p < 0.001 was considered statistically significant compared with control group.

 

Table 1: Effect of control, ranitidine and different extracts of Corchorus capsularis in pyloric ligation induced ulcer model.

 

Figure 2: Sample of dissected stomach and formation of ulcer in aspirin induced ulcer model. (a) Control, (b) ranitidine, (c) CCLD, (d) CCHD.

All values are shown as mean ± SEM, n = 6. ***p < 0.001 was considered statistically significant compared with control group.

 

Table 2: Effect of control, ranitidine and different extracts of Corchorus capsularis in aspirin induced ulcer model.

 

Aspirin is an acetyl salicylic acid derivative, nonselective COX-2 inhibitor, used as an analgesic.  Due to non-selectivity on COX pathway it also induces ulcer in the patient and has side effect. Because of blockage of COX-1 pathway prostaglandin synthesis get inhibited and causes damage in the mucosal layer of stomach. By using this phenomena aspirin has been used as an ulcer inducer in preclinical antiulcer screening since long time.

 

In aspirin induced ulcer model the control has shown ulcer index 12.50±0.17, in the other hand ranitidine, CCLD and CCHD have shown ulcer index 1.92±0.08, 6.42±0.22 and 4.50±0.09 respectively. Ranitidine is a standard drug has shown total acidity of 8.06±0.62, at the same time CCLD and CCHD have shown 12.23± 0.36 and 10.13±0.72 respectively and all of these three has shown significant level of decrease in total acidity while compared with untreated control i.e. 19.21±0.93 (Table 2).

 

Formation of ulcer is a complex mechanism and certainly correlate with mental condition. Any emotional and mental stress can cause acidity and ultimately form ulcer in the victim. In the stress induced ulcer model the animal has gone through a continuous stress of four hours by water immersion technique. As like previous models here also Corchorus capsularis in two different doses has shown its antiulcer activity in all the four parameters used namely ulcer incidence, ulcer index, total acidity and percentage of ulcer inhibition significantly while compared with the untreated control. It has also been seen that CCHD has shown certain level of better result than CCLD (Table 3), proves gradient response of Corchorus capsularis seeds extracts in dose dependent manner.

 

Corchorus capsularis has shown significant antiulcer activity in not only surgically induced, drug induced ulcer model but also in stress induced ulcer model that stands very important to establish its anti-ulcer activity scientifically and warrant for further study.

 

Figure 3: Sample of dissected stomach and formation of ulcer in stress induced ulcer model. (a) Control, (b) ranitidine, (c) CCLD, (d) CCHD.

All values are shown as mean ± SEM, n = 6. ***p < 0.001 was considered statistically significant compared to control group.

 

Table 3: Effect of control, ranitidine and different extracts of Corchorus capsularis in stress induced ulcer model.

 

CONCLUSION:

This study was designed to assess whether the chemical constituents of Corchorus capsularis could promote ulcer healing in experimentally produce ulcer in rat. The observation substantiated the use of Corchorus capsularis seeds in future for treatment of ulcer.

 

In this study the Corchorus capsularis seeds were extracted in soxhlet apparatus by using 70% ethanol. The extract was formulated and given orally, decreased the ulcer induce as well as the total acidity. Acidity is a parameter directly influence the chance of formation of ulcer.

 

Lipid peroxidation is an important process of several types of wound and ulcer²⁸. Any drug that inhibits lipid peroxidation¹¹ is believed to increase the circulation thereby preventing the cell damage, promoting DNA synthesis followed by reconstitution of broken wound or ulcer. The antioxidant property of Corchorus capsularis may be most responsible for its anti-ulcer activity.

 

Though several types of medications are also available for the treatment of ulcer still Corchorus capsularis can bring a new era in the treatment of ulcer which is easily available and affordable.

 

CONFLICT OF INTEREST:

The authors have no conflict of interest regarding this investigation.

 

ACKNOWLEDGEMENTS:

The authors would like to express their most sincerely appreciation to Dr. Arnab Samanta, Principal, Netaji Subhas Chandra Bose Institute of Pharmacy. Authors also acknowledge the support and help of all non-teaching staffs of Netaji Subhas Chandra Bose Institute of Pharmacy.

 

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Accepted on 05.02.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(12):5751-5756.