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Accelerated senescence of renal

tubular epithelial cells is associated


with disease progression of patients
with immunoglobulin A (IgA)
nephropathy
Liu, J., Yang, J., He, Y., Cai, G., Zhang, J., Lin, L., Zhan, J., , Xiao, H.(2012).
Accelerated
senescence of renal tubular epithelial cells is associated
with disease
progression of patients with immunoglobulin A
(IgA) nephropathy. Translational
Research,159(6), 454463.

Julie Nieva D. Sianquita


3Bio5
College of Science, Department of Biological Sciences
University of Santo Tomas

Disease: Immunoglobin A nephropathy (IgNA)

Bergers disease
Kidney disease that occurs when an antibody called Immunoglobin A (IgA) lodges
in the kidney.
Result in local inflammation that, over time, may hamper the kidneys ability to
filter waste, excess water and electrolytes from the blood.

Affecting Cells: Renal Tubular Epithelial Cells

epithelial cells, usually larger than granulocytes, contain a


large round or oval nucleus and normally slough into the
urine in small numbers.

with nephrotic disease the number sloughed is increased.

Suggested that its accelerated senescence may contribute


to the progression of IgNA

Studyblue.com

Image of a RTEC

Methodology
Renal Biopsy
(sample)
SA--Staining

Statistical Analysis
Nuclei stained with
DAPI

Stained with
fluorescein
isothiocyanate- or
Cy3-conjugated
IgG

Protein expression
of p16, p21, Col III
and FN

blocked and
incubated with
anti- -SMA or antiCol III antibody
together with anti
p16 or anti-p21
antibody

Methodology

SA--gal staining

Renal biopsy was obtained from patients

Fixed with cold acetone for 5 minutes

Washed with an acidic solution (pH 6.0) 3 times

Incubated with SA--gal staining solution for 12 hours at 37

Dried at room temperature

Mounted with neutral balsam

Protein expression of p16, p21, rabbit antihuman anticollagen(Col III) and


antifibronectin (FN)

Renal biopsy sections were processed with dewaxing, hydration and 3%


hydrogen peroxide treatment for 15 minutes at room temperature

Boiled in sodium citrate solution for 30 minutes then allowed to cool to room
temperature

Methodology

Rinsed in phosphate-buffered saline (PBS)

Blocked with goat serum for 15 minutes at room temperature

Incubated with polyclonal primary antibody overnight

Rinsed with PBS

Stained with horseradish peroxide-conjugated anti-mouse or antirabbit


immunoglobin G for 30 minutes at 37C

Rinsed with PBS

Incubated with diaminobenzidine (DAB) reagent

Double stained with hematoxylin and eosin (H&E)

Dehydrated with ethanol

Cleared with xylene

Mounted with neutral balsam

Methodology

To evaluate the association between p16 and p21 in RTECs and -smooth
muscle antibody (-SMA) and Col III in renal interstitium

Tissue sections were blocked and incubated with anti- -SMA (1:100 dilution) or
anti-Col III antibody (1:100 dilution) together with anti p16 (1:100 dilution) or
anti-p21 antibody (1:100 dilution) at 4 overnight

Rinsed with PBS

Stained with fluorescein isothiocyanate- or Cy3-conjugated IgG (1:50 dilution) for


60 minutes at 37C

Nuclei were stained with DAPI

Samples were mounted with glycerol and visualized under a confocal scanning
microscope

Results

The expression of senescence-associated markers, such as SA--gal, p21,


p16 were significantly in patients with grade I-II IgAN compared with normal
controls.

A linear correlation analysis revealed that the expression of senescenceassociated markers was associated significantly with blood pressure and
renal dysfunction but not with patient age, BMI, LDL cholesterol level, or 24h urinary protein value.
Percentages of p16 and p21- positive RTECs and SA--gal positivity were
associated closely with glomerular sclerosis, interstitial fibrosis, and vessel
lesion.

Conclusion

The results indicated that renal tubular epithelial cells (RTECs) in


immunoglobulin A nephropathy (IgAN) patients exhibited features of
accelerated senescence, which were unrelated to mechanisms associated
with normal aging.

Observations indicated that IgAN could induce the accelerated senescence


of RTECs, and cellular senescence might contribute to disease progression.
Insufficient supply of blood in renal interstitium might be associated with the
accelerated senescence or RTECs.
This study indicates that the accelerated senescence of RTECs might
contribute to the progression of IgAN by trigerring inflammatory response
and the deposition ofepeithelial cell matrix protein.