Effect of Methamphetamine on the Gut Epithelial Barrier Function

Poster Number

25

College

College of Arts and Sciences

Department

Biology

Faculty Mentor

Dr. Sabita Roy, University of Minnesota, Twin Cities

Abstract

Methamphetamine(METH) is a highly abused and addictive stimulant, with an estimated 35 million users worldwide. METH usage elicits a plethora of adverse consequences in the body including the brain, the circulatory system, heart, and liver. Recent studies by our lab suggests that METH increases bacterial translocation within the liver, lung, and mesenteric lymph node. Bacteria can be translocated by two mechanisms—an imbalance within the microbiome or an impaired epithelial barrier. Therefore, in the present study, we analyzed the effect of METH on epithelial barrier function using two cell lines—IEC-6 and CaCo-2. We hypothesize that METH will modulate the barrier function by inducing cell death, and disrupting the epithelial tight junctions. We found that high dosage METH at 100 micromolar causes a significant reduction in cell viability at 24h and 48h in IEC-6 and CaCo-2 cells using CCK8 assay, however, low dosage of METH at 10 micromolardoesn’t have the same effect. We also found that 10 micromolarMETH caused a significant increase in caspase-3 activity in IEC-6 cells. Furthermore, using Electric Cell Impedance Sensing(ECIS) and immunofluorescence we show a disruption in barrier integrity. In summary, this study shows that METH treatment in high doses disrupts epithelial barrier function by modulating tight junction integrity and epithelial cell viability.

Previously Presented/Performed?

Summer Undergraduate Research Symposium, University of Minnesota, Twin Cities, August 2015
Annual Biomedical Research Conference for Minority Students (ABRCMS), Seattle, Washington, November 2015

Grant Support?

Supported by grants from the National Institute of Health

Start Date

22-4-2016 2:15 PM

End Date

22-4-2016 4:15 PM

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Apr 22nd, 2:15 PM Apr 22nd, 4:15 PM

Effect of Methamphetamine on the Gut Epithelial Barrier Function

Richardson Ballroom

Methamphetamine(METH) is a highly abused and addictive stimulant, with an estimated 35 million users worldwide. METH usage elicits a plethora of adverse consequences in the body including the brain, the circulatory system, heart, and liver. Recent studies by our lab suggests that METH increases bacterial translocation within the liver, lung, and mesenteric lymph node. Bacteria can be translocated by two mechanisms—an imbalance within the microbiome or an impaired epithelial barrier. Therefore, in the present study, we analyzed the effect of METH on epithelial barrier function using two cell lines—IEC-6 and CaCo-2. We hypothesize that METH will modulate the barrier function by inducing cell death, and disrupting the epithelial tight junctions. We found that high dosage METH at 100 micromolar causes a significant reduction in cell viability at 24h and 48h in IEC-6 and CaCo-2 cells using CCK8 assay, however, low dosage of METH at 10 micromolardoesn’t have the same effect. We also found that 10 micromolarMETH caused a significant increase in caspase-3 activity in IEC-6 cells. Furthermore, using Electric Cell Impedance Sensing(ECIS) and immunofluorescence we show a disruption in barrier integrity. In summary, this study shows that METH treatment in high doses disrupts epithelial barrier function by modulating tight junction integrity and epithelial cell viability.