INTRODUCTION:

Lesions that develop on the stomach's inner lining or in the duodenum—the first segment of the small intestine—are known as peptic ulcers¹. An illness known as peptic ulcer disease causes ulcers, or open sores, to develop in the lining of the GI tract². The term "peptic" describes its connection to digestion. Pepsin, the main digesting enzyme the stomach produces, is the basis for the word³. In many nations, a significant portion of the population suffers from peptic ulcers, also known as ulcus pepticum⁴. This ailment is known as Amla pitta in Ayurveda. A common, persistent, and recurrent problem in humans, peptic ulcers can occasionally be fatal and have an unclear cause⁵. Helicobacter pylori infection and nonsteroidal anti-inflammatory drug use are the primary causes in the United States⁶. Peptic ulcer disease symptoms include epigastric pain (especially pain relieved by meals or Antacids are and pain that causes midnight awakenings or between meals), appetite loss, and weight loss⁷. Endoscopy should be performed very away on elderly individuals and those with alarm signs that point to a problem or cancer⁸. The prevalence of H. pylori infection is considerable and contributes to the rising morbidity and mortality rates associated with PUD⁹. It is a leading cause of global hospitalizations, with approximately 301,400 deaths occurring worldwide in 2013¹⁰. Among patients in Sub-Saharan Africa who underwent surgery for PUD, 86% had duodenal ulcers and 14% had gastric ulcers. The main complications necessitating surgery included perforation (355 cases), bleeding (7%), obstruction (30%), and chronic cases (28%), with an overall fatality rate of 5.7%¹¹. Gastric ulcers and duodenal ulcers are the primary varieties of peptic ulcers. Gastric ulcers are found in the stomach, whereas duodenal ulcers are located in the first segment of the small intestine (the duodenum)¹². Peptic ulcers can also occur in the oesophagus, though this is less common. A significant number of individuals with peptic ulcers may not show any signs of the condition¹³. When symptoms are present, they might manifest as a dull or burning sensation in the abdomen. This discomfort can differ from person to person; for some, it may worsen between meals and during the night, while others might feel an increase in pain after eating¹⁴. Other possible symptoms include a sensation of fullness or bloating, regular belching, heartburn, and feelings of nausea¹⁵. If peptic ulcer disease (PUD) is not identified and managed quickly, it can result in significant complications¹⁶. One of these complications is upper gastrointestinal bleeding, which can be a common and potentially life-threatening issue¹⁷. Another possible consequence is gastric outlet obstruction, where food cannot move from the stomach into the small intestine. In more severe instances, an ulcer may perforate the walls of the stomach or intestines, allowing gastric content to spill into the abdominal cavity and potentially causing peritonitis¹⁸. Penetration represents another grave complication, involving the ulcer extending into neighbouring organs like the pancreas or liver. Furthermore, persistent PUD may elevate the risk of gastric cancer, especially in cases linked to chronic Helicobacter pylori infections¹⁹. Therefore, swift diagnosis and proper treatment are essential to avert these repercussions. The process of diagnosing a peptic ulcer usually includes an assessment of medical history, a physical examination, and several tests²⁰.

MATERIALS AND METHODS:

Plant Collection and Authentication:

The bark of M. parvifolia was gathered from a location near Mazar Chauraha at the NCC Girls' Hostel in Prayagraj during February. The Botanical Survey of India (BSI) in Prayagraj, Government of India, has verified the plant. The bark of Mitragyna parvifolia was gathered, and the specimen received certification from Mr. Vinay Ranjan, Scientist-E and Head Officer at the BSI Central Regional Centre, located at 10 Chatham Lines, Prayagraj 212002.

Preparation of Extract:

After gathering the plant bark, it was first cleaned with tap water and then again with double-distilled water. After that, it was allowed to air dry for fifteen to twenty days at room temperature. After drying, the bark was ground into a powder with a grinder and kept for research in a dry, clean, airtight bottle.

Soxhlet Extraction:

The extraction process of Mitragyna parvifolia bark using Soxhlet was performed with a hydroalcoholic solvent. For this procedure, 50 grams of coarsely powdered and shade-dried Mitragyna parvifolia bark were placed in a thimble made from Whatman filter paper, which was then inserted into the main chamber of the Soxhlet extractor. A hydroalcoholic solvent was prepared using ethanol and distilled water in a 70:30 (v/v) ratio, and 1000 mL of this solvent was poured into a 1-liter round-bottom flask connected to the extractor²¹. The Soxhlet apparatus was properly assembled with a condenser at the top, and the flask was set on a heating mantle to start the extraction process²². Upon heating, the solvent began to evaporate, condensed in the condenser, and then flowed into the extraction chamber containing the plant material. This setup allowed for the continuous and repeated washing of the bark powder with fresh solvent, thus ensuring effective extraction of phytoconstituents. The extraction was carried on for approximately 6 to 8hours until the siphon tube indicated that a colourless solvent was present, signalling that the extraction was complete. After completion, the extract was gathered and condensed under low pressure using a rotary evaporator. The obtained concentrated extract was refrigerated and kept in an airtight container for later phytochemical and pharmacological testing²³.

Experimental animals:

Albino rats (200–250g) of both sexes were acquired from the Chakraborty Enterprises located in Kolkata, West Bengal and used for acute toxicity studies following OECD guidelines 423. The Institutional Animal Ethical Committee (IAEC) reviewed and approved all experimental protocols (IAEC approval no. SIP/IAEC/006/05/25).

Fig 1. Preparation of Plant Extract

Preliminary Phytochemical Screening:

The preliminary phytochemical screening aims to identify bioactive phytoconstituents in Mitragyna parvifolia bark, including Tannins, alkaloids, and flavonoids, which may contribute to its anti-ulcer activity, using traditional chemical tests, qualitative tests, and a combination of these methods.

Acute Toxicity Studies:

The study will use 20 male albino rats in good condition to test the acute toxic effect of bark extract from Mitragyna parvifolia in accordance with OECD Guideline 423. There will be five groups of rats, each consisting of four rats. Toxicological symptoms will be noted as the extract is given at varying dosages. The LD50 of the root extract will be determined by comparing fatality counts with the recommended values.

Antiulcer Effect by Pylorus ligation:

For this investigation, healthy adult albino Wistar rats of both sexes weighing between 200 and 250g were employed. The animals were kept in typical laboratory settings, with a 12-hour light-dark cycle, a temperature control of 25±2°C, and a relative humidity of 50–60%. They have free access to drinking water and a typical pellet diet. The rats were given free access to water and fasted for a full day before to the experiment.
Five groups of four rats each were created from the animals. Oral distilled water was given to the control group. Pylorus ligation was carried out under light anesthesia in accordance with normal protocols one hour after therapy delivery. After four hours, the animals' stomachs were gently removed and they were mercifully slaughtered. Gastric contents were collected to measure gastric juice volume, pH, and free and total acidity. The excised stomachs were then gently washed, opened along the greater curvature, and examined for ulcer formation¹⁹.

Table 1: Grouping of animals for evaluating the in-vivo antiulcer activity of hydroalcoholic extracts of Mitragyna parvifolia

S. No.	Name of Groups	Treatment	Animals	Total no. of animals
1.	Normal Control	Vehicle (Distilled Water, Orally)	Wister Rat	4
2.	Disease Control	Pylorus Ligation only (no treatment)	Wister Rat	4
3.	Test Dose Ⅰ	Extract 200 mg/kg (Low dose)	Wister Rat	4
4.	Test Dose Ⅱ	Extract 400 mg/kg (High Dose)	Wister Rat	4
5.	Standard Group	Ranitidine 150 mg/kg	Wister Rat	4
Total Animal Required				20

Evaluation Parameters of Ulcers:

After the animals were sacrificed, the stomachs from all experimental groups were carefully removed. The collected samples were then used to evaluate the ulcer index, percentage of ulcer protection, gastric juice volume, pH, total acidity, and the percentage inhibition of ulcer formation, following the prescribed experimental methods²⁴.

Fig 2. Excised Rat Stomach for Pharmacological

Ulcer Index (UI):

The degree and severity of stomach ulcers are assessed using a numerical metric called the Ulcer Index (UI). It is determined by calculating the average number of ulcers seen in each animal, the percentage of animals in an experimental group who have ulcers, and the intensity of ulceration based on ulcer size and depth²⁵. A standard scale is used to grade the severity of ulcers:

0 signifies normal stomach mucosa

1 signifies superficial ulcer,

2 signifies deep ulcer,

3 signifies perforations.

Ulcer Index = (UN + US + UP) × 10^-1

In this equation, UN represents the average number of ulcers observed per animal, US denotes the mean severity score of the ulcers, and UP indicates the percentage of animals exhibiting ulceration.

Percentage Inhibition of Ulcer:

Evaluates the safeguarding impact of a particular medication or experimental drug.

UI(control) – UI (test)

% Inhibition = ---------------------------------------- x 100

UI(control)

Determination of Gastric volume and pH:

After being gathered in graduated tubes, the stomach contents were centrifuged for ten minutes at 1000rpm. Following centrifugation, the clear supernatant's volume was measured. One milliliter of gastric juice and one milliliter of distilled water were combined from this supernatant, and the pH of the mixture was measured with a pH meter. While a higher pH implies less acidity and is thought to be beneficial for ulcer healing, a lower pH shows higher gastric acidity, which is linked to an increased risk of ulcer formation^24,26.

Determination of Total Acidity:

A 50mL conical flask was filled with a 1mL sample of gastric juice and an equal volume of distilled water. After adding two drops of phenolphthalein indicator, 0.01N sodium hydroxide was used to titrate the mixture until a stable pink endpoint was seen. After noting the amount of NaOH used, the total acidity was computed and represented in mEq/L using the proper formula²⁵.

V_NaOH x N x 100mEq/L

Acidity=---------------------------------------
0.1

Where, V = Volume

N = Normality

RESULTS:

Phytochemical Screening:

The According to several studies, the ethanolic extract of Mitragyna parvifolia bark contains flavonoids, alkaloids, phenols, and tannins, according to the preliminary phytochemical study.

Pyloric ligation induced gastric ulcer:

The study found that ulcer induction effectively reduced stomach acid output, ulcer index, and stomach volume, while the hydroalcoholic Mitragyna parvifolia bark extract significantly reduced ulcer index and stomach secretions.

Table 2: Effects of Ethanolic Bark Extract of Mitragyna parvifolia on Pylorus Ligation-induced Ulcers

Groups	Gastric Volume (ml)	Free Acidity (mEq/L)	Total Acidity ((mEq/L)	Ulcer Index (Mean ± SEM)	% (Ulcer Protection)
Group 1- Normal Control (Normal Saline)	5.0 ± 0.20 ****	5.0 ± 0.15 ****	12.0 ± 0.25 ****	0.5 ± 0.05 ****	00.00
Group 2-Disease Control	8.5 ± 0.30	16.0 ± 0.40	40.0 ± 0.60	6.5 ± 0.20	00.00
Group 3-Test 1 (MP 200mg/kg)	7.2 ± 0.25 ****	12.0 ± 0.35 ****	30.0 ± 0.50 ****	3.8 ± 0.15 ****	41.5
Group 4-Test 2 (MP 400mg/kg)	6.0 ± 0.20 ****	9.0 ± 0.30 ****	23.0 ± 0.45 ****	2.3 ± 0.10 ****	64.6
Group 5- Standard (Ranitidine 150mg)	5.5 ± 0.15 ****	8.0 ± 0.25 ****	20.0 ± 0.40 ****	1.8 ± 0.08 ****	72.3

Histopathological Analysis:

A histological examination of the lining of the stomach was carried out in order to validate the biochemical findings and assess the extent of mucosal damage for protection. The stomach tissues were removed following the sacrifice, sectioned, stored in 10% formalin, and stained with haematoxylin and eosin (HandE) for microscopic examination. Gastric tissues examined under a microscope showed clear histological differences between the experimental groups. There was no indication of ulceration, congestion, or inflammatory cell infiltration in the normal control group, which had intact stomach mucosa with a well-preserved epithelial lining and normally placed gastric glands²⁷. The ulcer control group, on the other hand, showed severe pathological abnormalities that significantly disrupted the gastric architecture, such as widespread mucosal ulcers, bleeding, submucosal edema, loss of epithelial cells, and intense inflammatory cell infiltration. Significant gastroprotection was achieved by treatment with the conventional medication ranitidine (150mg/kg), as shown by mostly intact glandular and epithelial tissues with relatively minor inflammatory alterations. At a dose of 200mg/kg, the extract-treated group demonstrated considerable mucosal healing, which was marked by decreased edema and inflammatory cell infiltration as well as partial epithelial layer regeneration, suggesting a protective effect. Remarkably, the extract administered at 400mg/kg resulted in nearly normal stomach histoarchitecture with little inflammatory alterations and minimal mucosal damage, similar to the group that received standard treatment²⁸.

Fig 4. Histopathological Analysis: (A) Normal Control (B) Ulcer Control (C) Standard (Ranitidine 150mg/kg) (D) Test Group Ⅰ (Extract 200mg/kg) (E) Test Group Ⅱ (Extract 400mg/kg)

DISCUSSION:

The hydroalcoholic bark extraction for Mitragyna parvifolia was evaluated in the current study using a pylorus ligation-induced stomach ulcer model including Wistar rats to determine its potential antiulcer properties and acute toxicity profile. The extract's potent gastroprotective qualities increased in a dose-dependent manner, according to the data. The 400mg/kg dose demonstrated ulcer inhibition comparable to that of the widely used drug ranitidine. The demonstrated antiulcer activity is caused by the phytochemical components identified during the initial screening, including flavonoids, alkaloid compounds, tannins, and phenolic chemicals. Flavonoids are compounds and tannins are well known for their cytoprotective and antioxidant properties; they protect the stomach mucosa by scavenging free radicals and reducing oxidative damage. The anti-secretory qualities of alkaloids, which reduce stomach acidity, may help promote the healing of ulcers. In the pylorus ligation model, gastric hypersecretion is a crucial factor in ulcer formation due to the increased production of the pepsin and hydrochloric acid. The hydroalcoholic extract of M. parvifolia significantly reduced stomach value, total acidity, and free acidity, suggesting both oral and antisecretory protective actions. The increase in pH levels in treated groups further supports the extract's capacity to reduce the harmful effects of acid from the stomach on the stomach lining. Histopathological results corroborated the biochemical data. The untreated ulcer control group displayed severe mucosal destruction, edema, or inflammatory infiltration, whereas therapy in an extract retained oral architecture, reduced inflammation, and enhanced epithelial regeneration, particularly at the 400 mg/kg dose. This preservation shows that the extract supports mucosal defense factors, possibly by boosting stomach mucosal blood flow and increasing mucus secretion. The acute toxicity research, conducted in compliance with OECD Guideline 423, demonstrated a broad safety margin and no evidence of toxicity or fatality at dosages up to 2000 mg/kg. This suggests that when an extract is administered at therapeutic levels, the likelihood of serious adverse effects is low. The gastroprotective qualities with M. parvifolia bark extracts are consistent with other research on medicinal plants with comparable phytochemical profiles that showed antiulcer effects via antioxidant, anti-inflammatory, and acid-suppressive pathways. The most recent research supports its traditional use in gastrointestinal disorders and emphasizes it as a promising natural substitute for chemical-based antiulcer drugs, especially in light of growing concerns regarding the long-term efficacy and resistance of conventional medicines such as protons pumps inhibitors and H₂-receptor antagonists. Ultimately, Mitragyna parvifolia hydroalcoholic bark extract exhibits strong, dependent on dose antiulcer effects with a good safety record, most likely due to a confluence of antioxidant, cytoprotective, and antisecretory mechanisms. To completely establish its efficacy in the management of peptic ulcer disease, more research is necessary, including the isolation of its active constituents, the clarification of specific molecular routes, and clinical testing.

CONCLUSION:

In a model of stomach ulcers caused by pylorus ligation in Wistar rats, the present study shows that the hydro alcohol-based bark extracts of Mitragyna parvifolia has strong and dose-dependent antiulcer activity. The higher dose (400 mg/kg) showed ulcer inhibition comparable to ranitidine, suggesting strong gastroprotective potential. The reported effects are most likely caused by bioactive phytoconstituents such as flavonoids, which are alkaloids, tannins, and phenols that might have cytoprotective, antioxidant, and antisecretory characteristics. Histopathological examinations confirmed the biochemical results, demonstrating that groups treated with extract had reduced inflammation and maintained mucosal architecture. Acute toxicity studies, that demonstrated a wide safety margin with no adverse effects at doses over 2000mg/kg, further indicate its potential for safe therapeutic application. These results demonstrate M. parvifolia potential as a natural alternative to prescription antiulcer drugs and validate its long-standing use for treatment of gastrointestinal disorders. Future studies should focus on determining the active ingredients, elucidating the precise mechanisms of action, and conducting clinical trials in order to demonstrate its efficacy and safety in humans.

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Received on 27.01.2026 Revised on 06.03.2026

Accepted on 08.04.2026 Published on 21.04.2026

Available online from April 24, 2026

Res. J. Pharma. Dosage Forms and Tech.2026; 18(2):121-127.

DOI: 10.52711/0975-4377.2026.00019

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