Current Symposium Poster

Effects of Trigonella Foenum-Graecum on Hypoglycemia in C. Elegans

By: Aditi Adapala, Mahathi Shankarram, Riya Patel, Srivar Chigurupati, Jishnuu Senthil Kumar, Pranav Rajan

Attempted Assays

Methods:

Abstract:

Total Content Assays

• Isolated compounds from fenugreek
• amino acids, alkaloids, flavonoids, saponins, phenolics
• Extraction methods
• Soxhlet reflux, Ultrasonic-assisted extraction, Liquid-liquid extraction

• Antioxidant assay

• Studies have shown that the development of diabetes mellitus is partly associated with oxidative stress and non-enzymatic protein glycation
• Assay can be used to identify compounds that offer antidiabetic activity

• MTT Assay

• The assay has been optimized and validated to rapidly assess the viability of C. elegans after drug treatment.
• C. elegans take up MTT and reduce it to the blue formazan, providing visual, qualitative, and quantitative assessment of viability.
• The assay can be used to quantitate the effects of different fenugreek extracts on the viability of C. elegans.

We have isolated bioactive compounds from fenugreek using the most effective extraction methods and quantified the phytoconstituent concentration via total content assays. From our extracts, we have attempted assays like nile red, pharyngeal pumping, and the antioxidant assay on the C. Elegans model throughout this project to help further understand our extracts and the potential they have against diabetes.

Total Phenolics Content Assay

Standard: Gallic Acid

Total Flavonoids Content Assay

Standard: Quercetin

Colorimetric Total Content Assays were performed for each extract with a Spectrophotometer and compared to a standard.

Total Alkaloids Content Assay

Standard: Berberine Chloride

Total Triterpenoid Content Assay

Standard: Diosgenin

Introduction:

In Vivo Attempted Assays

Conclusion:

• Diabetes Mellitus are a group of metabolic diseases that result in too much sugar in the blood
• Often caused by a lack of insulin or insulin resistance
• Current treatments for DM:
• Dietary management and physical activity
• insulin therapy
• oral medication (Metformin)

• Fenugreek is a medicinal plant used in cooking and medicine for several years
• Consists of many phytoconstituents including amino acids, phenolics, flavonoids, saponins, and alkaloids

• Quantification of each extract gave significant information of the contents of each phytoconstituent
• These studies can help further explore the roles of the compounds as inhibitors and/or treatments of other diseaseds
• Our results aren't consistent enough to form definite conclusions
• Trying to attempt new assays to measure antioxidant potential
• Reattempt assays we have done in the past to obtain consistent results
• Nile Red, Pharyngeal pumping

• Nile Red Assay
• Nile Red dye: Used to localize and quantify lipids, particularly neutral lipid droplets within cells
• Assay involved Nile Red staining of the lipids in hyperglycemic C. Elegans worms exposed to fenugreek extracts
• Measured if the lipid content was greater, smaller, or the same than the control (hyperglycemic worms not exposed to extracts)

• Pharyngeal Pumping Assay
• Feeding tendencies of C. Elegans controlled by pharynx, via rhythmic contractions and relaxations
• Induced hyperglycemia (high blood sugar) in the worms
• Assay tested effect of each fenugreek extract on the worms' pharyngeal pumping rates
• higher pumping rates = higher blood sugar (hyperphagia)
• Slowing pharyngeal contractions indicates hypoglycemic properties

Future Studies:

In terms of future studies, we hope to conclude our assays soon and publish our paper before researching fenugreek's properties in cancerous cells to determine its effects in treating cancer.

Acknowledgments:

We would like to thank:

• Mrs. Gayathri Renganathan
• Previous group members
• ASDRP lab staff
• ASDRP
• Olive Children Foundation

References :

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2. O. Trott, A. J. Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading, Journal of Computational Chemistry 31 (2010) 455-461 DOI 10.1002/jcc.21334

3. Ref. Dassault Systèmes BIOVIA, Discovery Studio Modeling Environment, Release 2017, San Diego: Dassault Systèmes, 2016.

4. The PyMOL Molecular Graphics System, Version 1.2r3pre, Schrödinger, LLC.

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