High-fat diet prevents the

development of
autoimmune diabetes in
NOD mice
NUTR 625
Ilana Fried
 Historical Perspective
● John Rollo and William
Cruickshank treated two patients
with glycosuria with a diet full
of vegetables, meat and fat.
○ One patient responded to the
diet
○ The other did not!
● Inconsistent response - higher
protein content in one of the
diets (glucose value of 58%)
Historical Perspective
● Rollo’s diet was further improved upon
in 1874 when Lauder Brunton published
this in the British Medical Journal,
regarding a low-carb diet

“Consisting of nitrogenous food, such as butcher-meat,

fish, eggs, and soups. Fat (which does not contribute
in the least to the formation of sugar) may be given in
all its forms, such as cream, butter, cheese, and oil.
Spinach, lettuce, and cresses may be freely used, but
celery and radishes only sparingly; while potatoes,
carrots, parsnips, turnips, peas, French beans, cabbage,
Brussels sprouts, cauliflower, broccoli, asparagus,
seakale, and fruit of all kinds, both fresh and
preserved, should be avoided,”
Historical Perspective

● The Allen Era (1914-1922)

○ Severe fasting to reduce blood
glucose
○ Control of calories
○ Low carb, high fat
○ Cause of death fell over 40%
● The “Michigan Diet” of Newburgh and
Marsh
○ Patients placed on an increasing
high fat diet (90g, 140g, 170g)
○ No adverse effects of the diet
○ Acidosis cleared up
○ Limiting protein and carbohydrates

Louis “Harry” Newburgh

Low carb + Very low carb diet: Type
2 diabetes
● Presented by Laura Saslow, PhD (School of
Nursing)
● ADA guidelines for type 2 diabetes - low
or very low carbohydrate diet
● Why is it recommended?
○ Lower glucose
○ Lower insulin
○ Weight loss
○ Lower blood pressure
○ Reduced inflammation
● Patients have been able to stop
glucose-lowering medication
 Introduction
● Type 1 diabetes is an autoimmune
disease where beta-cells are
destroyed and can’t produce
insulin
● Studies have shown that there is a
strong environmental component of
developing the disease
○ Infections
○ Diet
○ Gut microbiome
○ Vitamin D deficiency
● T1D has increased as childhood
obesity has increased
Q1
● Determine the effect of HFD-induced
obesity on the development of T1D in
genetically predisposed individuals
What is the purpose of ● Explore the relationship between a
this study? high fat diet and diabetes
● Is a high fat diet an environmental
trigger for diabetes?
 Q2
In the introduction,authors
mention opposing views and
previous conclusions regarding a
high fat diet and the
development of T1 diabetes. What
are these two views and what
have these studies concluded?
● View 1: HFD initiates and
accelerates Type 1 diabetes
○ Induces insulin resistance
○ Increases oxidative stress
○ Impairs insulin function
○ Induces beta-cell death
● View 2: HFD has protective effects
against the development of
diabetes
○ Stimulate beta cell
replication
○ Increase beta-cell mass
Q3
● Non-obese Diabetic Mice
● Develop spontaneous diabetes
What is a NOD mice? What with similar pathophysiology
is the purpose of using a to humans with T1D
NOD mice in this
particular study?
Q4 ● 4-week old NOD female mice
● Randomly separated into Standard
Chow Diet (SCD) or High Fat Diet
(HFD)
○ SCD - 21% F, 55% C, 23% P
Briefly describe the study ○ HFD - 44% F, 40% C, 15% P
design. ● Mice had free access to their specific
diet
● Body weight and blood glucose were
monitored weekly
● Mice were considered diabetic once
they had two random blood glucose
values >250 mg/dL on two separate
days
● Once declared diabetic, mice were
killed
Q5

Figure 1A shows the weight

gain in the SCD-NOD mice
and the HFD-NOD mice from 4
weeks to 40 weeks old. What
is the trend in this graph?
What could be a possible
explanation for this trend
in weight gain? ● Mice on HFD gained more weight
than the Mice on the SCD diet
● Fat is more calories than
carbohydrates and protein
● 9 kcal/g
Q6
Figure 1B focuses on
non-fasted blood glucose of
SCD-NOD mice and HFD-NOD
mice. Explain what is shown
in the graph and offer an
explanation for the sharp
increase at 14 weeks among
the SCD-NOD mice.
● Random blood glucose
measurements for HFD and SCD
mice
● Elevation in non-fasted blood
glucose in HFD-NOD mice in
weeks 6-10
● SCD-NOD mice had an increase
in blood glucose around week 14
● Loss of beta cell mass and
glucose intolerance in NOD mice
begins after 10 weeks
Q7

Figures 1D-1F
demonstrate the fasting
blood-glucose between 4
weeks and 15 weeks. What
are the graphs
demonstrating?

● (D,E,F): fasting blood glucose

between groups was pretty
similar between 4 weeks and 15
weeks.
 Q8
For the glucose tolerance test,
mice were injected with 2g/kg
dextrose and their blood glucose
was monitored a 0, 15, 30. 60 and
120 minutes at 4 weeks, 10 weeks
and 15 weeks. Based on figures
1G-1I, how does a HFD impact
glucose tolerance compared to the
SCD?
● (G): Glucose levels were similar between HFD
and SCD mice at 4 weeks
● (H): Impaired glucose tolerance among HFD
mice at 10 weeks
● (I): Glucose levels still higher among HFD
mice
Q9

While insulin resistance

and impaired glucose
tolerance were observed in
the HFD-NOD mice, what
surprising outcome was
observed? (Figure 2A)
● HFD mice had a higher
percentage of being diabetes
Reminder: *Mice were considered free compared to the SCD mice
diabetic once they had two random ● More SCD mice developed
blood glucose values >250 mg/dL on diabetes compared to the HFD
two separate days* mice
 ● Type 1 diabetes is
Q10 associated with
reduced beta cell
mass
What is the relationship
between beta-cell mass and ● Mice fed the HFD had a
type 1 diabetes? Take a significant increase in
look at figures 2D and 2E. beta-cell mass compared
What is happening with the to the SCD mice
beta-cell mass between the
SCD-NOD mice and the
HFD-NOD mice?

Reminder: *Beta cell mass was

evaluated at 10 weeks because this is
before significant loss of beta-cell ● Green - insulin
mass in NOD ● Red- glucagon
● Much more insulin and larger size in
40 week HFD mice
Q11
● To grade insulitis sections of the
Figure 3A shows the insulitis.
pancreas were stained
What does grade 1 – 4 indicate ○ Grade 1 - no islet associated
and how are the results mononuclear cell infiltrates
different between the HFD and ○ Grade 4 - invasion of the
the SCD? How does this relate to
the development of diabetes?
interior of the islet by immune
cells
● Less islet infiltration = less insulitis
● Patients with Type 1 diabetes: their
immune system attacks the islet cells
(infiltrates the islet cells) and kills
them
● Islet cells contain beta cells which
produce insulin
● T-regulatory cells ● To examine the
have been shown impact of
to suppress T-regulatory cells
immune response on the HFD-NOD
and decrease mice, they reduced
T-regulatory cells
autoimmunity
by using
● Increase immune
cyclophosphamide
regulation ● CP - has been
● More splenic shown to reduce
T-regulatory cells T-regulatory cells
in HFD-NOD mice ● Increased incidence
of diabetes
 Q12 ● 4 weeks: Types of
bacteria between
SCD and HFD mice
were similar
How did the HFD alter the
microbiome of the mice?
● 8 weeks: Large
increase in
bacteroidetes
among SCD mice

● 25 weeks: Increase
Bacteroidetes have been shown to increase risk for in verrucomicrobia
and developing T1D in humans

Previous study: Verrucomicrobia increased islet

T-regulatory cells and delayed onset of diabetes
Q13

● Potentially recommend a
How do you think the high fat diet for patients
results of this study who are predisposed to T1
will inform future diabetes.
dietary recommendations ● Patients who have a family
for people who may be history of T1 diabetes to
predisposed to diabetes? prevent onset.
 Study Conclusions
● While HFD-NOD mice gained weight, developed
decreased glucose tolerance and insulin
resistance, they were protected from developing
diabetes
● HFD-NOD mice protective factors in the study
○ Increased insulin secretion
○ Increased beta cell mass
○ Decreased insulitis
○ Altered stool microbiome
● Further studies on the mechanism of the high fat
diet in prevention of diabetes could lead to
interventions to prevent or delay T1D development
References
● Clark AL;Yan Z;Chen SX;Shi V;Kulkarni DH;Diwan A;Remedi MS;
“High-Fat Diet Prevents the Development of Autoimmune Diabetes in
Nod Mice.” Diabetes, Obesity & Metabolism, U.S. National
Library of Medicine,
https://pubmed.ncbi.nlm.nih.gov/34212475/#:~:text=Conclusions%3A%2
0This%20study%20shows%20that,regulatory%20cells%20and%20decreased%
20insulitis.
● Henderson G (2016) Court of Last Appeal - The Early History of the
High-fat Diet for Diabetes. J Diabetes Metab 7: 696. doi:
10.4172/2155-6156.100696
● UpToDate,
https://www.uptodate.com/contents/type-1-diabetes-and-diet-beyond-
the-basics.