Master of Science
Dr. Doug Quan
Ischemia reperfusion injury (IRI) occurs during transplantation and causes apoptosis and inflammation. The purpose of this research was to determine the effect of caspase-3, complement 3, and RelB gene silencing in the reduction of IRI using an in vitro model.
LLC-PK1 cells were used along with antimycin A for the in vitro IRI model. Prior to exposure to antimycin A, cells were transfected with caspase-3, C3, and RelB small interfering RNA (siRNA) alone or in combination and then analyzed.
The relative risk reduction of apoptosis in antimycin A treated cells with caspase-3 siRNA was 46.6% (p=0.019), RelB siRNA 42.8% (p=0.038), and complement 3 siRNA 13.9% (p=0.968). Combinations caspase-3 and RelB siRNAs showed significant changes, but were similar to transfection with caspase-3 and RelB alone.
Caspase-3 and RelB siRNA are effective at reducing apoptosis in an in vitro model of IRI and will be used in future large animal studies.
Zwiep, Terry M., "Treatment of Ischemia Reperfusion Injury with RNA Interference" (2015). Electronic Thesis and Dissertation Repository. 3422.