ABSTRACT
Diabetes mellitus is a persistent metabolic condition marked by elevated blood glucose levels due to compromised insulin secretion or functionality. The search for natural antidiabetic agents has gained attention due to their potential effectiveness and safety profiles. Sessuvium portulacastrum, a coastal plant, has been traditionally used for various medicinal purposes. This study investigates the antidiabetic potential of Sessuvium portulacastrum aqueous extract by analyzing its inhibitory effects on key enzymes involved in carbohydrate metabolism and exploring its molecular interactions with critical target proteins. The aqueous extract of Sessuvium portulacastrum was prepared and used for in vitro analysis. The reduced activity of the extract against α-amylase and α-glucosidase enzymes, crucial in glucose absorption and postprandial hyperglycemia, was assessed. Molecular docking techniques were employed to explore the potential interactions between active compounds in the extract and diabetes-related proteins, including BAX, GSK3β, and CADH. The study revealed significant inhibition of both alpha-amylase and alpha-glucosidase enzymes by Sessuvium portulacastrum aqueous extract, indicating its potential to reduce glucose absorption and postprandial hyperglycemia. Moreover, the molecular docking analysis demonstrated strong binding interactions between active compounds in the extract and key proteins involved in diabetes-related pathways, namely apoptotic pathways, glycogen synthesis, and cell adhesion. The findings of this study highlight the promising antidiabetic potential of Sessuvium portulacastrum aqueous extract. Upcoming research should get an attention on isolating and characterizing the active compounds responsible for these effects on antidiabetic therapies from natural sources.
KEYWORDS: Antidiabetic, alpha-amylase, alpha-glucosidase, molecular docking, BAX, GSK3B, CADH, novelty, diabetes mellitus, Health and well-being
INTRODUCTION
Diabetes mellitus, often referred to as diabetes, is a chronic metabolic ailment distinguished by increased blood sugar levels attributed to either inadequate insulin synthesis or impaired responsiveness to insulin.[1] This condition poses a substantial worldwide health challenge, impacting a vast population. Despite progress in medical knowledge, managing diabetes remains complex, urging the exploration of supplementary and adjunctive treatment approaches.[2] In current years, there has been an increased interest in natural products as potential sources of anti-diabetic drugs. Sessuvium portulacastrum, commonly known as ‘sea purslane’ or ‘beach succulent’, is a coastal halophyte of traditional medicinal use in different regions.[3] Sessuvium portulacastrum has been used in traditional medicine for its purported anti-diabetic properties.[4] The plant is known to contain several phytochemicals, including flavonoids, polyphenols, alkaloids and terpenoids, which may contribute to its medicinal potential.[5]
The main objective is to investigate the anti-diabetic potential of Sessuvium portulacastrum through a comprehensive in silico and in vitro analysis. Through this multifaceted research, we gain knowledge of the molecular interactions between the bioactive compounds of the plant and key enzymes involved in glucose metabolism. In addition, the study evaluates the potential of the plant extract in mitigating hyperglycaemia in an experimental in vitro model.
MATERIALS AND METHODS
Preparation of plant material and aqueous extract
Sessuvium portulacastrum plants were collected from a coastal region known for the abundance of this species. The plant material was taxonomically authentic, and voucher specimens were deposited in the herbarium for future reference. Aerial parts of the plant were thoroughly washed to remove debris and dried in the shade at room temperature. The aqueous extract was prepared by soaking the ground plant material in distilled water at a fixed ratio (e.g., 1:10 w/v) for a specified period of time (e.g., 24 hours) with occasional shaking.
Enzyme inhibition assay
In vitro assays were conducted to determine the inhibitory activity of the aqueous extract against α-amylase and α-glucosidase. Commercially available enzymes were used for the assays. For the α-amylase inhibition assay, a standard substrate solution of starch was prepared, and the enzyme was pre-incubated with the extract. Afterward, the process was started by adding the substrate, and the reaction progress was monitored using a spectrophotometer. The α-glucosidase inhibition assay involved a similar procedure using p-nitrophenyl-alpha-D-glucopyranoside as the substrate.
Molecular docking
Molecular docking studies were conducted to investigate the potential interactions between the active compounds in the Sessuvium portulacastrum extract and target proteins (BAX, GSK3B and CADH). The 3D of the proteins was gained from the Protein Data Bank (PDB). The compound phenol, 2,4-bis (1,1-dimethylethyl)-6-methyl- was chosen for docking based on its high inhibitory potential. The chemical structure of the compound was obtained from PubChem, a chemical database. Molecular docking simulations were performed using molecular docking software, such as AutoDock, employing a grid-based approach.
Statistical analysis
The experiments were conducted in triplicate, and the results are presented as the mean ± standard deviation (SD). Statistical analysis was carried out using suitable software (such as GraphPad Prism), with significance assessed through one-way analysis of variance (ANOVA) followed by post-hoc tests.
RESULTS
α-amylase and α-glucosidase activity
The initial screening of the Sessuvium portulacastrum plant revealed the presence of various phytochemical compounds, with terpenoids being positively identified [Table 1]. The in vitro enzyme inhibition assay revealed that the Sessuvium portulacastrum aqueous extract exhibited significant inhibitory activity against both α-amylase [Table 2 and Figure 1] and α-glucosidase enzymes [Table 3 and Figure 2]. The inhibitory effects were concentration-dependent, with higher concentrations of the extract leading to greater enzyme inhibition. The extract demonstrated comparable or even superior inhibitory activity compared to the positive control acarbose, indicating its potential as a natural anti-diabetic agent.
Table 1.
Phytochemical screening
| S. No | Phytochemical | Presence/Absence |
|---|---|---|
| 1 | Phenols | ++ |
| 2 | Saponins | -- |
| 3 | Steroids | ++ |
| 4 | Tannin | ++ |
| 5 | Terpenoids | ++ |
Table 2.
α-amylase inhibitory activity
| Conc (μg/ml) | Mean (Std % inhibition) | Mean (Extract % inhibition) |
|---|---|---|
| 20 | 26.82 | 17.17 |
| 40 | 37.18 | 27.62 |
| 80 | 45.56 | 36.83 |
| 160 | 57.89 | 48.84 |
| 320 | 70.25 | 56.88 |
Figure 1.
Alpha amylase inhibitory activity. Represents the Antidiabetic Alpha amylase inhibitory activity (% of inhibition). Values are expressed in mean ± SD of 3 replication
Table 3.
α-glucosidase inhibitory activity
| Conc (μg/ml) | Mean (Std % inhibition) | Mean (Extract % inhibition) |
|---|---|---|
| 20 | 27.63 | 19.99 |
| 40 | 36.27 | 28.06 |
| 80 | 48.82 | 39.74 |
| 160 | 62.37 | 54.13 |
| 320 | 81.95 | 62.54 |
Figure 2.
Alpha glucosidase inhibitory activity. Represents the Antidiabetic Alpha glucosidase inhibitory activity (% of inhibition). Values are expressed in mean ± SD of 3 replication
Molecular docking analysis
Molecular docking results demonstrated that the drug phenol, 2,4-bis (1,1-dimethylethyl)-6-methyl- identified from the Sessuvium portulacastrum aqueous extract showed strong binding interactions with the target proteins BAX, GSK3B and CADH [Figure 3]. The docking scores and binding modes suggested favourable binding affinities, indicating the potential of this compound to modulate apoptotic pathways, glycogen synthesis, and cell adhesion, which are relevant to diabetes pathogenesis [Table 4].
Figure 3.
Molecular docking analysis of Phenol, 2,4-bis(1,1-dimethylethyl)-6-methyl ligand with targets IR, AKT and PI3K using PyRx software and 3D structure visualized using Biovia Discovery Studio
Table 4.
Molecular docking analysis
| Drug | Protein | Binding energy (Kcal/mol) |
|---|---|---|
| Phenol, 2,4-bis (1,1-dimethylethyl)-6-methyl | BAX | -6.4 |
| GSK3β | -63 | |
| CADH | -5.7 |
DISCUSSION
The findings from the in vitro enzyme inhibition assay indicate that the Sessuvium portulacastrum aqueous extract possesses potent inhibitory activity against α-amylase and α-glucosidase enzymes. These enzymes are crucial in the breakdown of complex carbohydrates into glucose, and their inhibition can lead to reduced glucose absorption and postprandial hyperglycaemia.[6,7,8] The concentration-dependent nature of the inhibitory effects suggests that the extract contains active compounds capable of modulating the enzymatic activities. The molecular docking results provide further insights into the potential mechanisms of action of Sessuvium portulacastrum extract. The strong binding interactions of the compound phenol, 2,4-bis (1,1-dimethylethyl)-6-methyl-, with the target BAX, GSK3B and CADH indicate that this compound may play a significant role in modulating key cellular pathways relevant to diabetes pathogenesis. The identified compound’s favourable binding affinities and interactions with these proteins suggest its potential as a multitarget agent in diabetes management.
CONCLUSION
The study reveals that Sessuvium portulacastrum aqueous extract exhibits significantly reduced activity against α-amylase and α-glucosidase enzymes, indicating its potential as a natural anti-diabetic agent. Moreover, its comparable or superior activity to the commonly used anti-diabetic drug acarbose suggests promising therapeutic properties. Molecular docking results further suggest a multitarget approach, with the drug phenol, 2,4-bis (1,1-dimethylethyl)-6-methyl-, interacting strongly with key diabetes-related proteins, potentially influencing apoptotic pathways, glycogen synthesis and cell adhesion. This study highlights the importance of exploring natural sources for novel anti-diabetic therapies and sets the stage for future research to isolate and characterize the active compounds.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
Acknowledgements
The authors would like to thank Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 600 077, India for providing infrastructure facilities to carry out this work.
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