| Clinical backgroundThe congenital hydronephrosis due to ureteral obstruction is common in clinicalpractice, occurred in1-2%of newborn. With the extention of obstruction time, therewill have a large number of collagen deposition and dead kidney cells. Renalinterstitial fibrosis has been increasing with the renal function decreased. Themechanism involved in the fibrosis of the obstructed kidney is unclear.The effectiveclinical treatment of renal interstitial fibrosis in children has not been found atpresent. MicroRNA (miRNA) as a non-coding small RNA fragments, many studieshave confirmed that they participated the level of post-transcriptional geneexpression in a wide range of regulation. There is a positive effect on celldifferentiation, proliferation, embryonic development, signal transduction in vivoand clinically relevant disease early diagnosis, treatment and prognosis. Danishpharmaceutical company has performed a LNA-antimiRNA-122Phase I clinicaltrials. Recent studies have found that the expression of miRNA-101reduced in themyocardial infarct location and the surrounding normal position, however, the cellswere transfected with miRNA-101,and they could inhibit the newborn rat fibroblastproliferation and collagen production, preventing myocardial fibrosis cells occur.Whether miRNA-101is also involved in renal interstitial fibrosis, and there is aworth exploring.So through the establishment of complete unilateral ureteralobstruction (CUUO) model in this experiment, to explore the relationship between the expression of interstitial fibrosis and miRNA-101of kidney, for the renalinterstitial fibrosis provide a new therapeutic approach.MethodsThirty male SD rats (8±1weeks old，weighted180±20g) were randomly dividedinto three groups including sham, CUUO7day and14day group. The rats wereintraperitoneal injection with10%chloral hydrate0.4ml/kg, after fixing the rat,disinfecting the skin and putting the towel, opening abdominal cavity along itsventral midline, pushing the intestinal tube to the right side by the swab, exposingthe left ureter, ligaturing the left ureter between the renal vein and the waist vein,suturing the skin, after disinfection and bandage, placing in the constant temperaturebox.The kidney samples were collected in7and14days repectively, and used fordetecting the expression of miRNA-101by realtime polymerase chainreaction(RT-PCR) and α-SMA and E-cadherin protein by western blot andImmunohistochemistry. The correlation between their expression were analyzed.ResultsImmunohistochemical chemical measurement results displayed α-SMA andE-cadherin were mainly expressed in renal interstitial.The expression of α-SMAgradually reduced in CUUO14day、7day and sham group, however, the expressionof E-cadherin gradually increased. The tubular lumen of CUUO14day and7daygroup are different from sham group, which show expansion obviously. RT-PCRresults showed that the expression of MiRNA-101in sham and CUUO7day groupwas (14.50±5.12) and (3.45±2.27) times higher than those of CUUO14day group,respectively;There was a significant difference among these three groups (P <0.05);The expression of α-SMA protein were1.22±0.48、0.50±0.06and0.21±0.03inCUUO14day、CUUO7day and sham repectively. Which was highest in CUUO14day group, lowest in sham group and had negative correlation with miRNA-101(r=-0.74，P<0.05).In contrast, the expression of E-cadherin were0.24±0.22、0.50±0.44and0.76±0.20in CUUO14day、CUUO7day and sham repectively waslowest in CUUO14day group, highest in sham group and had positive correlationwith miRNA-101(r=0.88，P<0.05). ConclusionsThe renal expression of miRNA-101decreased with the obstructive time andrelated with renal interstitial fibrosis proteins, which indicated that miRNA-101mayhad significant correlation renal interstitial fibrosis progression. |
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