Abstract: Ulcers are among the main problems that developing countries face, as has long been recognised. In developing countries, traditional healers provide primary healthcare to over 70% of the population. Ficus religiosa (F. religiosa) Miq. (Moraceae) is an important traditional medicinal plant used for spiritual purposes that is found across India, mostly around Indian temples. It is known by a number of slang terms, including peepal tree and arasa maram.

Methods:

The anti-ulcer efficacy of F. religiosa ethanolic extract (250 and 500 mg/kg body weight) was studied in animal models of ulceration produced by stress. Ranitidine was the norm. The histological analysis and ulcer area were utilised to evaluate F. religiosa's anti-ulcer properties. Furthermore, preliminary phytochemical screening and studies on the acute toxicity of F. religiosa were carried out. The results showed that the extract treatments prevented ulcer area and gastric secretion in a dose-dependent manner.

Results:

In albino mice, administration of 2,000 mg/kg extract did not result in any acute toxicity. The ethanolic extract of F. religiosa included flavonoids, according to a preliminary phytochemical investigation. Not even at comparatively high doses of the extract is harmful. The presence of flavonoids is most likely the cause of the anti-ulcer action.

Conclusion:

The plant extract also showed the presence of phenols, tannins, steroids, alkaloids and flavonoids, β-sitosteryl-D-glucoside, vitamin K, n-octacosanol, methyl oleanolate, lanosterol, stigmasterol, lupen-3-one.

Introduction:

It has long been acknowledged that one of the biggest issues facing developing nations is ulcers. Approximately 70% of people in developing nations receive their primary medical care from traditional practitioners [1]. Numerous plants have been examined and found to be effective in treating and managing ulcers, thanks to the ever-growing interest in alternative medicine.

Throughout India, Ficus religiosa (F. religiosa) Miq. (Moraceae) is a significant traditional medicinal plant that is primarily found close to Indian temples for spiritual purposes. It goes by several colloquial names, such as arasa maram and peepal tree. The plant's bark is rich in sugars, tannins, amino acids, steroids, flavonoids, and saponins, among other substances [2]. Traditional medicine makes use of this plant's bark and leaf extract. The plant's bark is used to treat various urogenital disorders such as hemorrhoids, ulcers, gastrohelcosis, leucorrhea, menorrhagia, and vaginal infections. Additionally, it has antibacterial, analgesic, anti-diabetic, anti-inflammatory, anti-burning, and anti-oxidant properties [3]. In this study, the ethanolic leaf extract of F. religiosa was investigated for its acute toxicological effects as well as its anti-ulcer (or ulcer-preventive) properties.

Materials & Methods:

Plant Material:

An F. religiosa plant that was growing in the Medicinal Plant Garden of Chebrolu Hanumaiah Institute of Pharmaceutical Sciences was selected and verified the plant's taxonomy identification by Dr. Pendyala Vijetha (Head of the Department of Pharmacognosy in Chebrolu Hanumaiah Institute of Pharmaceutical Sciences). After being sun-dried, the leaves were ground into a powder and kept in an airtight container. For extraction, 250 g of dried leaf powder was placed in the thimble of a soxhlet apparatus and allowed to sit there for 18 hours at 55 °C while being extracted with ethanol. Weighing was done after the extract was air dried at 25–30 °C. The extract was dissolved at various concentrations in 10 milliliters of phosphate buffer saline (PBS) for oral administration.

Phytochemical Screening:

Using techniques developed by Brain et al. [4], the presence of flavonoids, tannins, alkaloids, saponins, glycosides, and sterols/triterpenes in the ethanol extract of F. religiosa was assessed.

Animals:

Male Albino mice (20–25 g) and Wistar rats (180-200 g) were acquired from the animal house of Chebrolu Hanumaiah Institute of Pharmaceutical Sciences and housed in typical surroundings. They received free-flowing water and a diet high in rodents. The study was given the go-ahead by the institutional animal ethical committee and the experiments were conducted in compliance with CPCSEA guidelines.

Anti-ulcer activity of F. religiosa ethanolic extract

Ulcers brought on by stress (cold water immersion method):

The Wistar albino rats of both sexes were made to swim in a glass cylinder filled with water up to 35 cm above the floor and kept at 25 °C for three hours to cause stress ulcers. The animals were divided into three groups, each with six animals, and fasted for twenty-four hours before the experiment. Groups 3 and 4 received 250 and 500 mg/kg, p.o. of F. religiosa ethanolic extract, respectively; Group 2 received 50 mg/kg, p.o. of ranitidine as the standard control. Group 1 received 1.0 mL/kg, p.o. as the vehicle control. The animals were given four hours to swim in cold water following their medication treatment. Following this, the animals were put to death using a large dose of anesthetic ether. Using a dissecting microscope (10×), the stomachs were opened along their greater curvature and checked macroscopically for gastric erosions. The volume of the gastric juice was measured after it was collected into centrifuge tubes and centrifuged for 10 minutes at 1,000 r/min. The gastric juice's pH was measured using a pH meter, and the gastric content's free and total acidity were examined. The area of the ulcer (UA) was computed. Calculations were made to determine the degree of protection provided to the animals by the various treatments [5,6].

Histopathology:

Following the collection of the stomach contents and measurement of the UA, small portions of the stomachs from each group were embedded in paraffin wax and fixed in 10% (v/v) formalin. Using standard techniques, sections with a thickness of 5 µm were cut in a microtome and then mounted on glass microscope slides. The sections were photographed and examined under a light microscope following hematoxylin-eosin staining [7].

Acute Toxicity Studies:

Male albino mice, n = 6, were divided into four groups to examine the acute toxicity of F. religiosa extract administration. After an overnight fast, ethanolic extract (50, 500, and 2,000 mg/kg) was given to the animals. As a control group, there was an equal volume of PBS given to each animal. For a full day, behavioral changes in the animals were observed both before and after administration. For a maximum of 14 days, the treated animals were monitored for any indications of toxicity [8].

Statistical analysis:

The mean ± SEM is used to express the results. One-way ANOVA was used to calculate the statistical difference between means, and Dunnett's post hoc test (Del/was) was used to compare and analyze the data with a significance level of P>0.05.

Results:

One of the most effective stress models for causing ulcers in rats was water immersion stress [9]. The animal experienced both physiological and emotional stress from the model. We looked at the ethanolic extract from F. religiosa leaves' anti-ulcer properties in an animal model of cold stress. The present study's results demonstrated the anti-ulcer activity of the ethanolic extract from F. religiosa leaves, as demonstrated by the significant inhibition of ulcer formation induced by stress-induced ulcer models (Table 1). Rats given standard medication ranitidine showed a significant change in gastric secretion volume, whereas animals treated with 500 mg/kg of F. religiosa leaf extract showed no change. When the stomachs of the animals that did not receive treatment with F. religiosa or ranitidine were examined under a microscope, they revealed complete ulceration (Figure 1). Nonetheless, animals given ranitidine and F. religiosa ethanolic extract at doses of 250 mg/kg and 500 mg/kg showed protection against ulceration (measured in UA).

Table – 1:

Anti-ulcer activity of F. religiosa ethanolic extract.

The mean ± SEM is used to represent the gastric volume and UA of the treated and control animals. Significant differences exist between the values in the column denoted by different letters (one-way analysis of variance, P<0.001). Asterisk (*) values indicate significant differences (P<0.001) from the ulcer control. It is not relevant.

Microscopic Visualization of Ranitidine treated and F. religiosa-treated Ulcers

Figure – 1

Ethanolic extract from the leaves of F. religiosa with anti-ulcer properties.

A rat with an ulcer control showing a larger ulcer area; B: A rat's stomach after receiving ranitidine; C: Rats' stomachs after receiving extract 250 mg/kg; D: Rats' stomachs after receiving 500 mg/kg; E: Rats' stomachs without ulcer control; F: Rats' stomachs after receiving ranitidine; G: Rats' stomachs after receiving 250 mg/kg of F. religiosa extract showing normal mucosa with small strophic gland mild hyperplasia and no edema; H: Animals given 500 mg/kg of F. religiosa leaf ethanolic extract displaying normal mucosa with mild hyperplasia and mild edematous submucosa.

The histopathology of the stomach revealed that, like the animals treated with ranitidine, the animals that received 500 mg/kg of extract had a significant reduction in the formation of gastric lesions and submucosal edema. Photomicrographs showed hemorrhagic erosion, discontinuity in the epithelium cell lining, and severe submucosal damage in the ulcer control animals' mucosa. Rats given ranitidine showed normal mucosa, a small strophic gland, mild hyperplasia, and no edema. In a similar vein, the mucosa of animals given ethanolic extracts of F. religiosa at doses of 250 mg and 500 mg showed mild hyperplasia but otherwise appeared normal. However, in the latter, a somewhat edematous submucosa was observed. The ethanolic extract of F. religiosa caused mild CNS depression for a few hours following treatment, according to acute toxicological investigations (Table 2). Still, even up to 14 days after the dosage of 2,000 mg/kg, there were no signs of toxicity or death.

Table 2

Discussion & Report: 

The phytochemicals and flavonoids found in the extract may be crucial in preventing ulcers, even though the exact mechanism is unknown. Similar results with the F. arnottiana methanolic extract were obtained in our previous study, which we have reported [10]. The presence of proteins, carbohydrates, tannins, flavonoids, and phenolic compounds is confirmed by the phytochemical screening. Flavonoids are among the compounds that are particularly interesting because of their reported anti-ulcerogenic activity and ability to protect the stomach. This study clearly shows that the leaf extract of F. religiosa has strong anti-ulcer properties in animal models. When compared to reference medications ranitidine, it exhibits gastric antisecretory properties. Not even at comparatively high concentrations is the extract toxic. The presence of flavonoids is most likely the cause of the anti-ulcer activity. To further investigate and characterize the compounds included in the extract's biological activity, more research is being done.

Applications:

Recent years have seen a significant increase in interest in natural medicine, which is based on a conventional understanding of the pharmacological qualities of plants. Extract from F. religiosa exhibits excellent anti-ulcer properties. The findings point directly toward the creation of a possible novel medication for the treatment of ulcers.

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Author Contribution:

• Author 1: Design, Interpretation of Data and Documentation of Work
• Author 2: Collection of Plant and its Phytochemical Screening
• Author 3: Acute Toxicity Studies and Statistical Analysis

Acknowledgement:

We express our special thanks of gratitude to the management, staff, and students of Chebrolu Hanumaiah Institute of Pharmaceutical Sciences for providing us with all the necessary facilities for this Research.

Funding:

There is no funding from any external source. We authors equally arranged all the expences in this work.

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