Effect of Bcl-xL and Glucolipotoxicity on

Mitochondria Metabolism
Emily Kamma, Imelda Suen, and Grace Zhao, Faculty of Science, University of British Columbia
Mentor: Alexis Shih, Luciani Lab, Child & Family Research Institute

Introduction

Hypothesis
The overexpression of Bcl-xL decreases mitochondrial metabolism under conditions of
glucolipoxicity.

Normal-WT

High sugar / high fat diet HGHF-WT

(20% fat, 4% sugar)
Normal Bcl-xL present

2) At endpoint, perform tests with:

Seahorse mitochondria stress test kit to
measure mitochondrial function
High sugar / high fat diet HGHF-Bcl-xL MitoSOX Red reagent to measure amount
of reactive oxygen species
(20% fat, 4% sugar)
TUNEL assay to measure apoptosis
Overexpressed Bcl-xL
Diagram 2. 2 months old transgenic mice are used, prepared as in a related
Pancreatic -cells will be isolated 6 months after treatment for assessment.

experiment.3

increased oxidative stress. High levels of Bcl-xL reduces ROS by decreasing

mitochondrial activity and beta-cell death. However, high levels of Bcl-xL also increases
blood glucose levels, which negatively impacts the body. Although elevated glucose
fff
levels are damaging, decreased oxidative stress
and beta-cell death could improve

350

300

HGHF-WT
HGHF-Bcl-xL

Our study focuses on the regulation of mitochondrial metabolism in blood glucose

Normal-WT

levels. However, storage of glucose in muscle and tissue can also influence blood

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200

glucose levels and should be studied.

150

ATP
Production

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Summary

HGHF Diet - WT HGHF Diet - Bcl-xL Normal Diet - WT

Overexpression

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40

60

Time (min)

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100

120

Figure 2. ATP Production. (A) WT mice on a HGHF diet had lower ATP production than
normal diet mice, indicating the HGHF diet affected metabolism. HGHF-Bcl-xL mice
showed a further decrease in ATP production, suggesting lower mitochondrial
metabolism than the WT. (B) Mitochondriafffv
stress kit test revealed a similar decrease
in ATP production as Panel A.
A. HGHF-WT

Glucolipotoxicity

Oxidative Stress

Decreased
Glucose
Metabolism

Mitochondrial
Apoptosis

B. HGHF-Bcl-xL
Figure 3. MitoSOX Live Cell
Fluorescence Imagery.
Fluorescence imagery revealed a
greater presence of reactive
oxygen species (ROS) produced
by pancreatic mitochondria in
wild type cells compared to
overexpressed Bcl-xL mice cells.
The red colour indicates the
presence of ROS.

Bcl-xL
Overexpression

Prevents Beta
Cell Apoptosis
Decreased
Mitochondria
Metabolism

Increased Blood
Glucose Level

Decreased
Insulin
Secretion

Acknowledgements

Adapted from https://tools.lifetechnologies.com/content/sfs/gallery/high/g002499.jpg

60

% Cell Apoptosis by TUNEL

Assay

Normal chow diet

(5% fat, 1% sugar)
Normal Bcl-xL present

Methodology
1) Every week, perform a glucose tolerance
test (IGPTT) after fasting mice 6 hours
prior to testing

conditions of high fat and sugar, more ROS than normal is produced, resulting in

of Bcl-xL required for long-term beneficial effects.

Experimental Methods
Group

production. At low concentrations, the cell can neutralize these harmful species. In

Adapted from (Zhou et al., 2000)

K+

Diagram 1. ATP production leads to channels

closing and voltage-gated Ca2+ channels opening.
Increase in Ca2+ concentration signals for insulin
secretion (Maechler and Wollheim, 2001).

Reactive oxygen species (ROS) are produced when oxygen is reduced during ATP

pancreatic health over time. Further research is needed to determine optimal amounts

OCR (pmol/min)

Our research aims to study how Bcl-xL

affects metabolism specifically in
pancreatic beta-cells under conditions
of glucolipotoxicity in vivo.

Figure 1. Intraperitoneal Glucose

Tolerance Test (IPGTT). IPGTT
analysis of HGHF diet mice after 6
months showed higher blood
glucose levels in Bcl-xL
overexpressed mice than in WT
mice, suggesting impaired glucose
metabolism.

ATP Production

Diabetes is a metabolic disease that affects more than 360 million worldwide. Diabetic
patients have high blood sugar due to both insulin resistance and failure of the pancreas
to produce sufficient insulin, a hormone which maintains normal blood sugar levels.
Mitochondria is important in coupling metabolism with insulin secretion (Diagram 1).
The presence of high blood glucose and free fatty acid synergistically promotes oxidative
stress, beta-cell death, and decreased insulin secretion. This condition is termed
glucolipotoxicity.1 The cellular pathways involved in beta-cell failure still remains
unclear. Recent research has found that Bcl-xL, a protein localized primarily in the
mitochondria, reduces beta-cell apoptosis (programmed cell death) and affects
mitochondrial metabolism.2 However, there are no studies investigating the effect of
Bcl-xL on pancreatic beta-cells under
the conditions of glucolipotoxicity.

Treatment

Discussion

Anticipated Results

Figure 4: TUNEL Assay.

Percentage of cell apoptosis was
determined using TUNEL Assay.
Overexpression of Bcl-xL
reduced the number of cell
deaths in pancreatic beta-cells.

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We would like to thank Alexis Shih for her guidance and support, and the Undergraduate Research
Opportunities (URO) club for their support throughout this process.

References
1 Kim

10

JW, Yoon KH. Glucolipotoxicity in pancreatic -Cells. Diabetes Metab J. 2011, 35:44450. doi: 10.4093/dmj.2011.35.5.444.

2 Luciani

DS, White SA, Widenmaier SB, et al. Bcl-2 and Bcl-xL suppress glucose signalling in pancreatic -cells. Diabetes 2013, 62:170182. doi: 10.2337/db11-1464.

HGHF Diet - WT

HGHF Diet - Bcl-xL

Overexpression

Normal Diet - WT

3 Zhou

YP, Pena JC, et al. Overexpression of Bcl-xL in -cells prevents cell death but impairs mitochondrial signal for insulin secretion. Am.
J. Physiol. 2000, 278:340-355.