Objective To evaluate the changes in the expression of adaptor
protein containing pleck-strin homobgy domain,
phosphotyrosine-binding domain and a
leucine zipper motif 1(APPL1)during renal
fibrosis in a
mouse model of renal
ischemia-
reperfusion(I∕R)
injury.
Methods Twenty-four
male C57BL∕6
mice,
aged 8 weeks, weighing 20-25 g, were divided into 2 groups(n=12 each)using a random number table sham operation group(S group)and renal I∕R group.The model of renal I∕R
injury was established by clipping the bilateral renal pedicles for 30 min followed by
reperfusion in group I∕R.Six
mice were selected at 2 days of
reperfusion, and venous
blood samples were collected for
determination of
serum concentrations of
blood urea nitrogen and
creatinine.The
animals were then sacrificed, the renal specimens were obtained for microscopic examination of tubular
necrosis with a
light microscope, and the damage to the renal tubules was scored using a semi-quantitative
method.Six
mice were sacrificed at 14 days of
reperfusion, and the renal specimens were obtained for assessment of the degree of renal
fibrosis(using picric
acid-sirius red
staining) and for
determination of the expression of
collagen type 1,
fibronectin and α-
smooth muscle actin in renal tis-sues(by
Western blot or
immunofluorescence method). At 2 and 14 days of
reperfusion, the expression of APPL1 in renal
tissues was detected by
Western blot and the expression of APPL1
mRNA in renal
tissues by
real-time polymerase chain reaction. Results Compared with group S, the
serum concentrations of
blood u-rea
nitrogen and
creatinine, scores of renal tubular damage and degree of renal
fibrosis were significantly in-creased at 2 days of
reperfusion, the expression of
collagen type 1,
fibronectin and α-
smooth muscle actin in renal
tissues was up-regulated at 14 days of
reperfusion, and the expression of APPL1
protein and
mRNA was up-regulated at 2 and 14 days of
reperfusion in group I∕R(P<0.05). Conclusion Up-regulated expression of APPL1 may be involved in the process of renal
fibrosis in a
mouse model of renal I∕R
injury.