Exploring the Anthelmintic and Antidiabetic Effects of Rosarin in Preclinical Studies Using Zebrafish and Earthworm Models (Pheretima posthuma).

Abstract

Background: Rhodiola rosea is a traditional medicinal plant that has been found to possess several beneficial properties, including the ability to mitigate cardiac ischemia-reperfusion damage, reduce blood lipid levels, prevent thrombosis, and exhibit antiarrhythmic effects.

Objective: This study aims to evaluate the potential of rosarin, a key compound derived from the root of Rhodiola rosea, in treating diabetes mellitus using a zebrafish model and in exhibiting anthelmintic (worm-expelling) activity using the Indian earthworm (Pheretima posthuma).

Methods: The study design utilizes an experimental approach, incorporating both zebrafish (Danio rerio) and earthworms (Pheretima posthuma) as subjects for testing. The zebrafish were randomly assigned to different experimental groups, including control and treatment groups (e.g., hyperglycemia induction and comparison with Metformin). The zebrafish were studied for a duration of 4 days, during which the glucose concentration was gradually increased. Zebrafish were housed in controlled aquatic environments with daily water changes and hyperglycemia in zebrafish was induced by gradually increasing the glucose concentration, starting with 50 mM for four days while closely monitoring their health and survival. The body weights, blood glucose levels and histopathological studies were noted and compared with the standard drug Metformin. Liver enzymes such as alkaline phosphatase (ALP), Aspartate aminotransferase (AST), and Alanine aminotransferase (ALT) derived from homogenate supernatants of fish viscera were determined using an autoanalyzer. Earthworms were collected from moist soil and randomly assigned to receive varying doses of test Rosarin or the standard drug albendazole in Petri dishes. Observations included changes in color, thickness, diameter, paralysis, and time to death. Dunnett's test was used to evaluate the statistical significance, followed by one-way ANOVA.

Results and discussion: Zebrafish (Danio rerio), three-month-old (500-1000 mg) and Pheretima posthuma (14 cm) were used for this research. The results confirm that the rosarin glycoside at 50 mg/ml showed significant anti-diabetic activity by decreasing blood glucose levels (82.1 ± 0.5 mg/dl) with p<0.001, 95% CI (81.628- 82.572) limits and body weights (2.0 ± 0.047 g) when equated with diabetic control (Blood glucose levels= 135 ± 3.14 mg/dl and body weights =13.4 ± 0.11 g). ALT, AST and ALP levels significantly decreased in the rosarin group when equated to diabetic control. The anti-diabetic effect of rosarin is comparable with standard Metformin (50 mg/ml). In anthelmintic activity, rosarin (75 mg/ml) significantly decreased the length of the worm (9.5 ± 0.36 cm), time of paralysis (22 ± 0.76 minutes) and time of death (40 ±0.76 minutes). Albendazole (50 mg/ml) is used as a standard drug. The study employed one-way ANOVA to compare the means of various experimental groups, followed by Dunnett's test for post-hoc analysis to evaluate the differences between the treatment groups and the control group.

Conclusion: The results of the current research indicate that rosarin has significantly reduced the blood glucose levels in zebrafish and decreased the time of paralysis and death in earthworms, suggesting its antidiabetic and anthelmintic activity and hence it is further recommended as an ideal candidate for therapy of diabetes and worm infestations.