The Effects of a High

Protein Diet on
Development of Chronic
Kidney Disease and
Contributing Factors in
Jennifer Wihlborg
Male & Female Wistar Rats PI: Dr. Alireza Jahan-Mihan
 Research Assistants:
Sarah Funk Leila Ninya
4 French Students:
Kea Schwartz Madison Borland
Galle Edjoa-Mintcha
Tyler Gehm Ashley Denisar
Manon sacr
Tatyana Kimble Jillian Friederich
Lumineau Mlodie
Jessica Lindamood
Victoria Brown
Rachel Wakefield Houdelot Antoine
 Importance of the Kidneys
Waste removal
Urea, uric acid, creatinine, etc.

Fluid and electrolyte balance

Hormone release
Controls erythropoiesis
Bone health
 Background
Chronic kidney disease (CKD) is a condition characterized by a gradual loss of
kidney function over time.
CKD is a major health burden with the estimated prevalence of 8-16% worldwide.1
An association between high protein intake and decreased renal function has been
established through both clinical and animal studies.2-6
However, whether high protein diets alter renal function in a sex-dependent manner is
still unclear.
The mechanisms by which high protein diets may influence kidney function differently in
males and females are still elusive and need further study.
 High Protein Intake
High protein diets have been utilized widely in both preventive and therapeutic
approaches against obesity.
Proteins support lean mass by maintaining muscle mass and by stimulating protein
synthesis.1
Proteins are a more satiating macronutrient compared with fat and carbohydrates
(cal/cal).3
The average daily protein consumption in adults in the United States is far above
recommended daily allowance (0.8 gr/kg body weight) as suggested by national
dietary guidelines:
The net daily protein intake of males and females are 101.9 grams and 70.1 grams
respectively, which is more than double of the recommended amount.7
 Proteins Negative Effects on Renal Function
Decreases urinary citrate and pH while increasing creatinine
levels, kidney weight, renal mesangium (the membrane
supporting the adjoining glomerular capillaries) and glomerular
tufts.
This can directly lead to nephrolithiasis and decreased renal
function.8-10
May also deteriorate kidney function indirectly by worsening the
severity of hypertension and proteinuria
Major contributing factors in the etiology of renal disease.11
 Findings in Previous Studies
A recent study examined sex differences in the rate of loss of kidney
function and development of structural damage in ageing human and rats.16

Suggest that males are more vulnerable than females.

The protective effect of estrogen by stimulation of nitric oxide (NO)

production is proposed as potential mechanism.12

NO deficiency has been linked with chronic kidney disease progression (in
animal models).2
 Renal Disease Parameters
Blood urea nitrogen (BUN): detects elevated waste levels in the bloodstream
Normal range: 7-20 mg/dl

Serum Creatinine: determines how much creatinine is in the bloodstream

Normal range: .8-1.4 mg/dl
Glomerular Filtration Rate (GFR): determines levels of renal function

Stage GFR Levels & Description12

Stage 1 (CKD I) >90 mL/min; normal or high GFR

Stage 2 (CKD II) 60-89 mL/min; mild decrease in GFR

Stage 3 (CKD III) 30-59 mL/min; moderate decrease in GFR

Stage 4 (CKD IV) 15-29 mL/min; severe decrease in GFR

Stage 5 (ESRD, CKD V) <15 mL/min, dialysis dependent

 Methodology:
30 Post-Weaned 30 Post-Weaned
Male Wistar Rats Female Wistar Rats
(Normal Diet) (Normal Diet)

Euthanasia x6 Euthanasia x6
(Week 4) (Week 4)

12 High Protein 12 Normal Protein 12 High Protein 12 Normal Protein

Final Euthanasia Final Euthanasia Final Euthanasia Final Euthanasia

(week 14) (week 14) (week 14) (week 14)
 Analyses
Body weight and food intake (weekly)
Systolic (SBP) and diastolic (DBP) blood pressure and pulse (weeks 1, 4, 8, 12)
Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT) (weeks 1, 4, 8, 12)
HOMA-IR, body composition, insulin, IL-6 (weeks 1 and 12)
Urinalysis (weeks 1, 4, 8, 12)
pH, creatinine, protein, BUN

Body weight and composition, renal weight, and blood samples (at time of euthanasia)
Budget
Product Amount Price
LabSand Bedding $650

Normal Diet 75 kg ($17/kg) $1,275

High Protein Diet 75 kg ($24.50/kg) $1,837.50

Rat Insulin ELSIA 1 kit $332

Blood Glucose Test Strips 450 strips $250

BCA Dual Range Protein Detection Kit 2 plate $195

Urea Nitrogen (BUN) Calorimetric Detection Kit 2 plate $275

Serum Creatinine Detection Kit 2 plate $210

Total: $5,025.50
 Questions?

And remember...
 References
1. Jha V, Garcia-Garcia G, Yang C, et al. Series: Chronic kidney disease: global dimension and perspectives. The Lancet. 2013;382:260-272.
2. Friedman AN. High-protein diets: potential effects on the kidney in renal health and disease. AM J Kidney Dis. 2004; 44(6):950-62.
3. Calvez J, Poupin N, Chesneau C, Lassale C, Tome D. Protein intake, calcium balance and health consequences. Eur J Clin Nutr. 2012; 66(3):281-95.
4. Addis T. The effect of some physiological variables on the number of casts, red blood cells and white blood cells and epithelial cells in the urine of normal
individuals. 1926, J Clin Invest 2, 417421.
5. Wilson HE. An investigation of the cause of renal hypertrophy in rats fed on a high protein diet. Biochem J 1933, 27, 13481356.
6. Hammond KA, Janes DN. The effects of increased protein intake on kidney size and function. J Exp Biol 1998, 201 (Pt 13), 20812090.
7. Fulgoni VL 3rd. Current protein intake in America: analysis of the National Health and Nutrition Examination Survey, 2003-2004. AM J Clin
Nutr. 2008;87(5):1554S-1557S.
8. Aparicio V, Nebot E, Aranda P, et al. High-protein diets and renal status in rats. Nutricion Hospitalaria. 2013;28(1):232-237.
9. Malhotra R, Cavanaugh K, Blot W, Ikizler T, Lipworth L, Kabagambe E. Higher protein intake is associated with increased risk for incident end-stage renal
disease among blacks with diabetes in the Southern Community Cohort Study. Nutr Metab Cardiovasc Dis. 2016 Dec;26(12):1079-1087
10. Kostogrys R, Franczyk-arw M, Malak E, Topolska K. Effect of low carbohydrate high protein (LCHP) diet on lipid metabolism, liver and kidney
function in rats. Environmental Toxicology and Pharmacology. 2015;39:713-719.
11. Shen Q, Xu H, Wei L, Chen J, Liu H. Basic nutritional investigation: Intrauterine growth restriction and postnatal high-protein diet affect the kidneys in
adult rats. Nutrition. 2011;27:364-371.
12. Baylis C. Sexual dimorphism in the aging kidney: differences in the nitric oxide system. Nature Reviews Nephrology. 2009;5(7):384-396.
13. https://www.davita.com/kidney-disease/overview/symptoms-and-diagnosis/understanding-your-lab-work/e/4724