Effects Of TLR4 And UCP2 On Non-Alcoholic Steatohepatis Of The Liver

Emmaleigh Messer

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Kenneth Chavin, Division of Transplant Surgery, Medical University of South Carolina

Abstract

Mice with Toll-like receptor 4 (TLR4) induces the presence of UCP2 (uncoupling protein 2). UCP2 is found in great quantities in the hepatocytes of the Steatotic liver. UCP2 uncouples proteins in the mitochondria causing a decreased production of ATP. Steatotic mice lacking TLR4 have an increased survival rate because they do not produce as much, if any, UCP2 and therefore produce enough ATP. It was confirmed that TLR4 follow the JNK pathway to stimulate UCP2 growth in earlier experiments. During my six weeks, we genotyped the mouse colonies to confirm the presence or absence of TLR4 and UCP2 based on which gene was knocked out. We used Real-time PCR to find the presence of TNF-alpha, TNF-beta and CCL2, which are inflammatory cytokines. We also isolated hepatocytes by the perfusion and removal of the mice liver to be treated with LPS. The cells were cultured over a 24 hour time period and then treated with LPS, a bacteria commonly found in the gut. The RNA of the hepatocytes was then isolated and quantified using a nanodrop technique. The series of experiments showed an increase of NDUFB8 and Drp1, both of which show mitochondrial dysfunction in the lard-fat diet fed mice. The mice lacking TLR4 did not show any change, because the LPS was not accepted.

Start Date

4-11-2015 9:15 AM

End Date

4-11-2015 9:30 AM

COinS
 
Apr 11th, 9:15 AM Apr 11th, 9:30 AM

Effects Of TLR4 And UCP2 On Non-Alcoholic Steatohepatis Of The Liver

Mice with Toll-like receptor 4 (TLR4) induces the presence of UCP2 (uncoupling protein 2). UCP2 is found in great quantities in the hepatocytes of the Steatotic liver. UCP2 uncouples proteins in the mitochondria causing a decreased production of ATP. Steatotic mice lacking TLR4 have an increased survival rate because they do not produce as much, if any, UCP2 and therefore produce enough ATP. It was confirmed that TLR4 follow the JNK pathway to stimulate UCP2 growth in earlier experiments. During my six weeks, we genotyped the mouse colonies to confirm the presence or absence of TLR4 and UCP2 based on which gene was knocked out. We used Real-time PCR to find the presence of TNF-alpha, TNF-beta and CCL2, which are inflammatory cytokines. We also isolated hepatocytes by the perfusion and removal of the mice liver to be treated with LPS. The cells were cultured over a 24 hour time period and then treated with LPS, a bacteria commonly found in the gut. The RNA of the hepatocytes was then isolated and quantified using a nanodrop technique. The series of experiments showed an increase of NDUFB8 and Drp1, both of which show mitochondrial dysfunction in the lard-fat diet fed mice. The mice lacking TLR4 did not show any change, because the LPS was not accepted.