The Effect of Verapamil on P-Glycoprotein Expression In Marin-Darby Canine Kidney (MDCK) Cells

School Name

Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Melissa Moss, University of South Carolina

Abstract

Alzheimer’s disease is affecting over five million people around the world. The disease is caused by a build-up of amyloid plaques in the brain. These plaques are made of protein fragments derived from an amyloid precursor protein (APP), which is present in the membrane of neurons. In an average brain, these plaques are generally transported out of the brain by P-glycoprotein (P-gp), but with Alzheimer’s disease the plaques continue to build up. P-gp is a transport protein that is known to transport toxins in and out of the blood stream, in the kidney as well as the brain. In previous research, it was shown when cells were treated with Verapamil there was a greater expression of P-gp in the cell. Verapamil is a hypertensive drug; and therefore associated with blood flow in the vascular system, as well as might have a role to play with P-gp expression in transportation. The goal of this project was to gather preliminary data regarding P-gp expression in Marin Darby Canine Kidney (MDCK) cells that are treated with Verapamil, a drug. This was achieved by a) Caspase Assay and b) XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) Cell Proliferation Assay. The Caspase Assay showed that the Verapamil was not as toxic as the TNF-α. The Caspase Assay, however, did not yield reliable results. The XTT Assay, however, showed that the cells were still alive and metabolizing when treated with Verapamil. Based on these results a Western Blot will be done for P-gp in presence and absence of Verapamil.

Start Date

3-25-2017 11:59 PM

Presentation Format

Written Only

Group Project

No

COinS
 
Mar 25th, 11:59 PM

The Effect of Verapamil on P-Glycoprotein Expression In Marin-Darby Canine Kidney (MDCK) Cells

Alzheimer’s disease is affecting over five million people around the world. The disease is caused by a build-up of amyloid plaques in the brain. These plaques are made of protein fragments derived from an amyloid precursor protein (APP), which is present in the membrane of neurons. In an average brain, these plaques are generally transported out of the brain by P-glycoprotein (P-gp), but with Alzheimer’s disease the plaques continue to build up. P-gp is a transport protein that is known to transport toxins in and out of the blood stream, in the kidney as well as the brain. In previous research, it was shown when cells were treated with Verapamil there was a greater expression of P-gp in the cell. Verapamil is a hypertensive drug; and therefore associated with blood flow in the vascular system, as well as might have a role to play with P-gp expression in transportation. The goal of this project was to gather preliminary data regarding P-gp expression in Marin Darby Canine Kidney (MDCK) cells that are treated with Verapamil, a drug. This was achieved by a) Caspase Assay and b) XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) Cell Proliferation Assay. The Caspase Assay showed that the Verapamil was not as toxic as the TNF-α. The Caspase Assay, however, did not yield reliable results. The XTT Assay, however, showed that the cells were still alive and metabolizing when treated with Verapamil. Based on these results a Western Blot will be done for P-gp in presence and absence of Verapamil.