Project Title

Effects of Nictotinamide Riboside and Beta-Hydroxybutyrate on C. elegans Lifespan

Authors' Affiliations

Jeffery Peters, Department of Biomedical sciences, College of Medicine, Easy Tennessee State University, Johnson City, TN Patrick C. Bradshaw , Department of Biomedical Sciences, College of Medicine, East Tennessee State University, Johnson City, TN. Jeffery Peters and Patrick C. Bradshaw, , Department of Biomedical Sciences, College of Medicine, East Tennessee State University, Johnson City, TN.

Location

Ballroom

Start Date

4-12-2019 9:00 AM

End Date

4-12-2019 2:30 PM

Poster Number

48

Faculty Sponsor’s Department

Biomedical Sciences

Name of Project's Faculty Sponsor

Dr. Patrick Bradshaw

Type

Poster: Non-Competitive

Classification of First Author

Undergraduate Student

Project's Category

Biochemistry

Abstract Text

The vitamin B3 precursor nicotinamide riboside (NR) and the ketone DL-body beta-hydroxybutyrate (BHB) are two of the most promising natural compounds yet identified for the treatment of aging and aging-related diseases. NR increases nicotinamide adenine dinucleotide (NAD) levels to activate sirtuin protein deacetylases and BHB is an anti-aging calorie restriction (CR) mimetic. Caenorhabditis elegans is a 1 mm long nematode worm used as a model system to study aging with a mean lifespan of roughly 2-3 weeks depending upon the exact temperature of culture. NR and BHB have previously been shown to increase lifespan when administered to C. elegans by roughly 20%. However, the effect on lifespan when both compounds are added together has not yet been studied. It is hypothesized that when added together the effect on lifespan will be slightly larger than when either compound is given alone, due to the activation of complementary signaling pathways. The results will help determine if humans could benefit from taking both compounds simultaneously as most signaling pathways that regulate lifespan are conserved from nematodes to humans. For these experiments cultures of mixed age C. elegans nematodes were first treated with alkaline-bleach to kill adult worms leaving only eggs that are protected by their thick eggshell. Using this protocol isolated eggs are age-synchronized to within 9 hours of each other. The eggs were then transferred into E. coli-permeable, but nematode impermeable 8 micron cell culture inserts placed in twelve well microplates with roughly 25-40 eggs per insert. 1.35 mL of liquid S-media containing 9 x109 E. coli cells/mL as food was added to each well. Microplates were shaken to provide culture aeration. After three days, the worms reached adulthood and 0.4 mM fluorodeoxyuridine (FUdR), a DNA synthesis inhibitor, was added to prevent C. elegans egg-laying to maintain age-synchrony. Every Monday, Wednesday, and Friday for the approximate 4 weeks of the lifespan experiments the worms were counted under a microscope and the culture media and bacteria replaced. Results were analyzed using Kaplan-Meier survival curves and Log-rank analysis. Results indicated that individual treatment with BHB or NR or both combined increased lifespan in the two trials performed thus far. Another trial is currently underway, and results will be analyzed after it is completed to determine if the combined treatment has a greater benefit on lifespan then either individual treatment. Future studies could also be performed to determine if either NR or BHB can further extend the lifespan of animals given rapamycin, a TOR kinase inhibitor, another promising anti-aging therapeutic.

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Apr 12th, 9:00 AM Apr 12th, 2:30 PM

Effects of Nictotinamide Riboside and Beta-Hydroxybutyrate on C. elegans Lifespan

Ballroom

The vitamin B3 precursor nicotinamide riboside (NR) and the ketone DL-body beta-hydroxybutyrate (BHB) are two of the most promising natural compounds yet identified for the treatment of aging and aging-related diseases. NR increases nicotinamide adenine dinucleotide (NAD) levels to activate sirtuin protein deacetylases and BHB is an anti-aging calorie restriction (CR) mimetic. Caenorhabditis elegans is a 1 mm long nematode worm used as a model system to study aging with a mean lifespan of roughly 2-3 weeks depending upon the exact temperature of culture. NR and BHB have previously been shown to increase lifespan when administered to C. elegans by roughly 20%. However, the effect on lifespan when both compounds are added together has not yet been studied. It is hypothesized that when added together the effect on lifespan will be slightly larger than when either compound is given alone, due to the activation of complementary signaling pathways. The results will help determine if humans could benefit from taking both compounds simultaneously as most signaling pathways that regulate lifespan are conserved from nematodes to humans. For these experiments cultures of mixed age C. elegans nematodes were first treated with alkaline-bleach to kill adult worms leaving only eggs that are protected by their thick eggshell. Using this protocol isolated eggs are age-synchronized to within 9 hours of each other. The eggs were then transferred into E. coli-permeable, but nematode impermeable 8 micron cell culture inserts placed in twelve well microplates with roughly 25-40 eggs per insert. 1.35 mL of liquid S-media containing 9 x109 E. coli cells/mL as food was added to each well. Microplates were shaken to provide culture aeration. After three days, the worms reached adulthood and 0.4 mM fluorodeoxyuridine (FUdR), a DNA synthesis inhibitor, was added to prevent C. elegans egg-laying to maintain age-synchrony. Every Monday, Wednesday, and Friday for the approximate 4 weeks of the lifespan experiments the worms were counted under a microscope and the culture media and bacteria replaced. Results were analyzed using Kaplan-Meier survival curves and Log-rank analysis. Results indicated that individual treatment with BHB or NR or both combined increased lifespan in the two trials performed thus far. Another trial is currently underway, and results will be analyzed after it is completed to determine if the combined treatment has a greater benefit on lifespan then either individual treatment. Future studies could also be performed to determine if either NR or BHB can further extend the lifespan of animals given rapamycin, a TOR kinase inhibitor, another promising anti-aging therapeutic.