Pharmacological evaluation of the anti-ulcer and anti-arthritic potential of ethanolic extract of acalypha indica linn

 Corresponding author: S. SENTHILKUMAR Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution License 4.0. Pharmacological evaluation of the anti-ulcer and anti-arthritic potential of ethanolic extract of acalypha indica linn S. SENTHILKUMAR 1, *, P. KUMAR SINGH 1 and C. KIRUBA RANI 2 1 Manipur, International University, Imphal, Manipur, India. 2 Department of Biochemistry, Vellalar College for Women, Erode, Tamil Nadu, India. World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 Publication history: Received on 29 March 2025; revised on 05 May 2025; accepted on 08 May 2025 Article DOI: https://doi.org/10.30574/wjbphs.2025.22.2.0471 Abstract The objective of present study to evaluate the anti-ulcer and anti-arthritic activity of ethanolic leaf extract of Acalypha indica. The anti-ulcer activity of ethanolic extract of Acalypha indica was investigated by pylorous ligation and indomethacin induced gastric ulcer in rats. The cause of ulceration in patients is mainly due to hyper secretion of gastric acid and pepsin. Acalypha indica plant extract some of the most attractive sources of new drugs and have been shown to produce promising results in the treatment of gastric ulcers. Rheumatoid arthritis is a systemic auto immune disease characterized by articular inflammation that eventually leads to the destructions of joints. The present work is aimed to evaluate the anti-arthritic effect of plant Acalypha indica extract by Freund’s Complete Adjuvant (FCA) induced arthritis in rats. Medicinal plants possess numerous active phyto constituents that are responsible for several biological activities. Acalypha indica is very important medicinal plant, to possess anti-ulcer and anti-arthritic activity. Keywords: Indomethacin; Pylorous Ligation; Acalypha Indica; Freund’s Complete Adjuvant; Anti Arthritic; Ethanolic Extract; Helicobacter Pylori 1. Introduction Peptic ulcer is a gastro intestinal disorder due to an imbalance between the aggressive factors like acid, pepsin, Helicobacter pylori and defensive factors like bicarbonate secretion, prostaglandins, gastric mucus, innate resistance of the mucosal cell factors [1]. Normally peptic ulcers develops when aggressive factors overcome the defensive factors are Helicobacter pylori, acid pepsin hyper secretion, non-steroidal anti-inflammatory drugs, sometimes idiopathic due to usage of tobacco, psychological stress, rapid gastric emptying and zollinger-ulcer Ellisson syndrome where there is a high and uncontrollable production of acid also leads to ulcer formation [2,3,4,5]. Synthetic drugs such as proton pumb inhibitors, H2 receptors, cytoprotectants, demulcents, anti cholinergies, antacids and prostaglandin analogues are used for the treatment of ulceration but these drugs produce several side effects. So herbal medicines are considered as better alternatives for the treatment of peptic ulcer [6]. For example (Omeprazole, lansoprazole) may cause nausea, abdominal pain, constipation, diarrhoea and H2 receptor antagonists (cimetidine) may cause gynaecomastia, loss of libido. Due to the occurrence of many side effects by use of synthetic drugs for many diseases, medicinal plants are considered as the main source of new drugs as they have less or no side effects. As herbal medicines are considered as safe for the treatment of ulcers with lesser adverse effects, economical, effective, relatively less toxic, extensive research is carried out in search for potent anti-ulcer agents of plant orgin[7,8]. Rheumatoid arthritis is an immune mediated inflammatory disease. Rheumatoid arthritis occurs when our immune system attacks the tissues near joints, this is due to release certain chemicals and enzymes that begin to eat away the cartilage and bones. Rheumatoid arthritis affects all the joints in the body, some forms of arthritis can also affect the World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 148 body’s internal organs. The symptoms of Rheumatoid arthritis include inflammation, pain, swelling and stiffness of the joints, it can also lead to deformity and disability of the joint in sever cases[9]. These are several causes for Rheumatoid arthritis these causes are unknown but some include genetic factor, family history, age, environmental factors, hormones, smoking etc[10,11]. The medicinal plants with phyto constituents which can be used in the treatment of Rheumatoid arthritis[12]. 2. Materials And Method 2.1. Chemicals All chemicals used including the solvents were analytical grade. Folin-ciocalteu phenol reagent 1,1-diphenyl-z-picryl hydrazyl (DPPH), polyvinyl polypyrro lidone (PVPP), ammonium acetate and 2,4,6 – tripyridyl-s-triazine (TPTZ) were purchased from merch co. (Germany). 2.2. Preparation of Plant Extract The Acalypha indica plant were shade dried at room temperature. The dried material was then homogenized to obtain coarse powder and stored in air-tight bottles for further analysis. The shade dried, powdered leaves were extracted with ethanol by hot extraction using soxhlet apparatus, collected and stored in a vial for further analysis. 2.3. Animals The experiment was carried out by male wister albino rats weighing (150-175g) and were procured from small animals breeding station, Mannuthy, Kerala, India. The animals were housed under standard conditions of temperature (23±10C), relative humidity (55±10C), 12h /12h light / dark cycle and fed with standard pellet diet (Pranav Agro Industries Ltd., Sangli, India) and water ad libitum. Animals described as tested were deprived of food for atleast 18h. but allowed free access to water. All the experimental procedures and used in the study were received by the Institutional Animals Ethics Committee (Reg.No.688/2/C-CPCSEA) and were in accordance with the guildelines of the CPCSEA. 3. Antiulcer activity 3.1. Sample Preparation Coarse powder from the shade dried plant material was exhaustively extracted with ethanol to yield a dark greenish semisolid residue. The dried extract was dissolved in distilled water right before use. 3.2. Indomethacin (IND) Induced Ulcers The experiment was performed according to the method of Djahanguiri Four groups of male wister rats (n=4) were fasted overnight prior to the start of the experiment and water ad libitum. The first group received distilled water, which the second group was treated with omeprazole (10mg kg-1 day-1 P.O.). Where as third to fourth groups were administrated with the ethanolic extract of present study plants (200 and 400 mg kg-1 day-1 P.O. respectively). On day third after 30 min of omeprazole and plant extract treatments, indomethacin (50 mg kg-1) suspended in 0.5% carboxymethy cellulose was given as a single oral dose to groups 2-4 to induce gastric ulcers. After 5h, the animals were sacrified with over dose of diethylether and each stomach was examined for ulcer index[13]. 3.3. Experimental Design • Group – I: Control which received distilled water orally. • Group – II: Omeprezole (10 mg kg-1 day-1 P.O.) • Group – III: Served as test sample which received ethanolic leaf extract of 200 mg kg-1 P.O. of Acalypha indica. • Group – IV: Served as test sample which received ethanolic leaf extract of 400 mg kg-1 P.O. of Acalypha indica. 3.4. Ulcer Index The stomach were removed and opened along the greater curvature, washed gently in normal saline and the mean ulcer index was calculated. The number of ulcer lesion were counted using a magnifying glass and the diameter of the ulcer was measured using a vernier calliper. Ulcer index was determined by following the scoring method [14]. World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 149 • Score 1: Maximal diameter of 1mm • Score 2: Maximal diameter of 1-2 mm • Score 3: Maximal diameter of 2-3 mm • Score 4: Maximal diameter of 3-4 mm • Score 5: Maximal diameter of 4-5 mm • Score 10: Maximal diameter of 5 mm and above • Score 25: A per forated ulcer The sum of the length (mm) of all lesions for each stomach was used as the ulcer index (UI), and the protection percentage was calculated from the following formula. 𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑃𝑟𝑜𝑡𝑒𝑐𝑡𝑖𝑜𝑛 =𝑈𝐼 𝐶𝑜𝑛𝑡𝑟𝑜𝑙 – 𝑈𝐼 𝑇𝑟𝑒𝑎𝑡𝑒𝑑 𝑈𝐼 𝐶𝑜𝑛𝑡𝑟𝑜𝑙 ×100 3.5. Anti-Arthritis 3.5.1. Procedure Wister Albino Male rats (150-200g) were divided into 5 groups of six animals each (n=6). Group I served as control. Arthritis was induced in rats by injecting 0.1 ml of 0.1% Freund’s Complete Adjuvant (FCA), (Sigma Aldrich, USA) into the sub planter region in the right hint paw of Group II-V was administrated with 200 mg kg-1 day-1 P.O. and 400 mg kg1 day-1 P.O. ethanolic leaf extract of Acalypha indica daily for 15 days [15]. 3.5.2. Experimental Design The plant extract treatment was administrated as follows for 5 days. • Group – I: Served as Control • Group – II: Freund’s Complete Adjuvant (FCA) into the sub planter region in right hind paw. • Group – III: Administered with indomethacin (10 mg kg-1 day-1 P.O.) daily • Group – IV: 200 mg kg-1 day-1 P.O. of Acalypha indica • Group – V: 400 mg kg-1 day-1 P.O. of Acalypha indica The increase in joint diameter was measured daily starting from day 1, by using vernier calibre. Percentage protection rendered by the plant extract is calculated using the formulate, Percentage Protection = Difference in Paw volume of inducedDifference in Paw Volume of Standard / or treated 𝐷𝑖𝑓𝑓𝑒𝑟𝑒𝑛𝑐𝑒 𝑖𝑛 𝑃𝑎𝑤 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑖𝑛𝑑𝑢𝑐𝑒𝑑 × 100 3.6. Statistical Analysis The data presented here are means ± SD of 6 rats in each group. The results were analysed using one-way analysis of variance (ANOVA) and the group means were compared by Duncan’s Multiple Range Test (DMRT) using Statistical Program For Social Sciences (SPSS Version 16.0) software for windows. The findings were considered statistically significant at P < 0.05[16]. 4. Results and discussion Gastric lesions were induced in rats by oral administration of Indomethacin (50 mg kg-1). Oral administration of ethanolic leaf extract of Acalypha indica registered a significant dose dependent decrease in the extent of gastric mucosal damage in indomethacin induced ulcer models (Figure I). The protective effect of the Acalypha indica extract at a dose level 400 mg kg-1 b.w. (Ulcer Score 13.95; 67.72% Protection) was slightly lower than that of the standard drug omeprazole (Ulcer Score 11.37; 72.44% Protection). Indomethacin induced depletion of gastric wall mucous formation has been significantly reduced by ethanolic leaf extract of Acalypha indica at the dose levels of 200 and 400 mg kg-1 b.w. Showing protection percentage as 48.13 and 67.72 respectively (Table 1 and Fig 1). The anti-arthritic activity of the ethanolic leaf extract of Acalypha indica (Tables 2 and 3) studied against FCA – induced arthritis is exhibited in Figure II. The tested plant drug extracts of Acalypha indica at two different concentrations (200 and 400 mg kg-1 b.w. P.O.) were administrated orally and the activity was compared with the standard drug Indomethacin (10 mg kg-1 day-1 P.O.). Sub planter injection of FCA (Freund’s Complete Adjuvent) in the rat hind paw led World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 150 to the development of arthritis which reached a peak edema on the 10th day of injection. The standard indomethacin seemed to inhibit this edema to an extent of 64.14%. Among the plant drugs used in the present study, percentage protection of Acalypha indica at a high dose was slightly higher (71.22%) and it was very close to the standard (72.20%) on the 15th day (Table 3 and Fig 2). Ethanolic extract of Acalypha indica found to contain alkaloids, glycosides, flavonoids, saponins, phenolic compounds and tannins as their phytoconstituents. These secondary metabolites play an important role to act as an anti microbial, anti-oxidant, anti-inflammatory, anti-ulcer and anti-arthritic activity in the presently investigated plant in Acalypha indica. Table 1 effect of ethanolic extract of acalyphaindica leaves on ulcer index of indomethacin induced gastric ulcer in ratsA Group Ulcer Score Percentage Protection (%) Induced 36.17α ± 0.98 - Standard 11.37d ± 0.72 72.44 Low dose (200mg kg-1b. w) 19.87b ± 0.36 48.13 High dose (400mg kg-1b. w) 13.95c ± 0.38 67.72 aValues are expressed as mean ± SD for 4 animals (n=4) significant at P < 0.05 level. One way ANOVA Followed by DUNCAN’S Multiple Range Test. ACALYPHA INDICA Figure 1 percentage protection of ethanolic extract of acalypha indica leaves on ulcer index of indomethac in induced gastric ulcer in rats A B World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 151 C D Figure 2 Effect of ethanolic leaf extract of acalypha indica leaves on ulcer index of indomethacin induced gastric ulcer in rats Table 2 antiarthritis activity of ethanolic leaf extract of acalypha indicaa Paw volume(mm) Days Control Induced Standard Low dose High dose 0 3.28 ± 0.16 3.15o ± 0.12 3.40p ± 0.08 3.34p ± 0.11 3.37p ± 0.14 1 3.28 ± 0.16 6.96n ± 0.24 6.60i ± 0.21 7.23i ± 0.43 7.25a ± 0.47 2 3.28 ± 0.16 7.45m ± 0.23 7.27f ± 0.14 7.29g ± 0.40 6.89b ± 0.55 3 3.28 ± 0.16 7.95l ± 0.02 7.83d ± 0.06 7.47d ± 0.39 6.67c ± 0.42 4 3.28 ± 0.16 8.60k ± 0.09 8.38a ± 0.09 7.53c ± 0.38 6.07d ± 0.28 5 3.28 ± 0.16 9.32j ± 0.05 8.16b ± 0.04 7.66a ± 0.35 5.96f ± 0.25 6 3.28 ± 0.16 9.92h ± 0.04 7.89c ± 0.05 7.60b ± 0.33 5.87g ± 0.26 7 3.28 ± 0.16 10.25f ± 0.09 7.73e ± 0.11 7.48e ± 0.32 5.71h ± 0.17 8 3.28 ± 0.16 10.77d ± 0.04 7.23g ± 0.06 7.37f ± 0.32 5.60i ± 0.16 9 3.28 ± 0.16 11.44b ± 0.08 6.89h ± 0.10 7.27h ± 0.33 5.48j ± 0.13 10 3.28 ± 0.16 11.85a ± 0.06 6.52j ± 0.12 7.18j ± 0.32 5.99e ± 0.11 11 3.28 ± 0.16 11.23c ± 0.07 6.15k ± 0.05 6.98k ± 0.34 5.31k ± 0.12 12 3.28 ± 0.16 10.73e ± 0.06 5.73l ± 0.06 6.77l ± 0.33 5.24l ± 0.13 13 3.28 ± 0.16 10.11g ± 0.04 5.43m ± 0.06 6.59m ± 0.35 5.22m ± 0.10 14 3.28 ± 0.16 9.85i ± 0.08 5.25n ± 0.08 6.40n ± 0.36 5.20n ± 0.09 15 3.28 ± 0.16 9.30k ± 0.08 5.11o ± 0.04 6.20o ± 0.36 5.14o ± 0.08 a Values are expressed as mean ±SD for 6 animals (n=6) significant at p<0.05 level. One way ANOVA followed by Duncan’s Multiple Range Test. Table 3 percentage protection of acalypha indica leaf extract against FCA induced arthritis at different days intervalsa Days Group Initial paw volume(mm) Final paw volume(mm) Difference(mm) Percentage protection (%) 5th day Control 3.28d ± 0.16 3.28e ± 0.16 - - Induced 3.15e ± 0.12 9.32a ± 0.05 6.17a ± - Standard 3.40a ± 0.08 8.16b ± 0.04 4.76b ± 22.85 Low 3.34b ± 0.11 7.66c ± 0.35 4.32c ± 29.98 High 3.37c ± 0.14 5.96d ± 0.25 2.59d ± 58.02 10th day Control 3.28d ± 0.16 3.28e ± 0.16 - - World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 152 Induced 3.15e ± 0.12 11.85a ± 0.06 8.70a ± - Standard 3.40a ± 0.08 6.52c ± 0.12 3.12c ± 64.14 Low 3.34c ± 0.11 7.18b ± 0.32 3.84b ± 55.86 High 3.37b ± 0.14 5.99d ± 0.11 2.62d ± 69.89 15th day Control 3.28d ± 0.16 3.28e ± 0.16 - - Induced 3.15e ± 0.12 9.30a ± 0.08 6.15a ± 0.19 - Standard 3.40a ± 0.08 5.11d ± 0.05 1.71d ± 0.09 72.20 Low 3.21c ± 0.13 5.64b ± 0.51 2.86b ± 0.33 53.50 High 3.23b ± 0.11 5.13c ± 0.36 1.77c ± 0.18 71.22 a Values are expressed as mean ±SD for 6 animals (n=6) significant at p<0.05 level. One way ANOVA followed by Duncan’s Multiple Range Test. A B C D E Figure 3 percentage protection of acalypha indica leaf extract against FCA induced arthritis at different day intervals World Journal of Biology Pharmacy and Health Sciences, 2025, 22(02), 147-154 153 Acalypha Indica Figure 4 Percentage protection of acalyphaindicaleaves extract against FCA induced arthritis at different days intervals 5. Conclusion Herbal medicinal products have a wide scope of assorted variety of multi-dimensional synthetic strctures in the on going occasions, the utility of normal items as natural capacity modifiers has impressive considerations. The botanical study evaluated the identification of 2,50,000 to 3,50,000 plant species over the plant. The curative properties of medicinal plant are due to presence of major group of active components which are mainly alkaloids, phenols, flavonoids, steroids, saponins, terpenoids, tannins and glycosides. Plants are served as major structural resources for traditional as well as modern medicinal system all over the world. The therapeutic potential of plants and plant products can be treated back to thousands of years ago. Acalypha indica possess the antimicrobial, anti-oxidant, anti-inflammatory, anti ulcer and anti arthritic activities. These findings justify the usefulness of these plant in the management and treatment of inflammation associated diseases like ulcer and arthritis. Compliance with ethical standards Disclosure of conflict of interest No conflict of interest to be disclosed. Statement of ethical approval If studies involve use of animal/human subject, authors must give appropriate statement of ethical approval. If not applicable then mention ‘The present research work does not contain any studies performed on animals/humans subjects by any of the authors’. References [1] Dashputre N.L. and Naikwade N.S. (2011). 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