miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1

Figure 9

Direct siRNA-mediated suppression of HSPD1 in the UUO kidney promoted oxidative stress despite miR-382 blockade. (a) MiR-382 abundance determined by real-time PCR measurement of the whole kidney RNA from the obstructed mouse kidneys. (b–e) Protein expression of Bax, HSPD1, and Trx was normalized to GAPDH in the obstructed kidneys treated with anti-miR and siRNA from the experiment above. (f) Representative images of kidney tissue with Sirius red staining and 3-NT immunofluorescence from animals undergoing UUO surgery and receiving a combination of HSPD1 or control siRNA (ureteral delivery) and anti-miR-382 or control anti-miR (intravenous delivery). 3-NT (green) and DAPI (blue) immunofluorescence from control and anti-miR382/HSPD1 siRNA-treated mice are displayed (). (g) The relative abundance of collagen was analyzed using Sirius red staining. (h) The positive-stained area of 3-NT was quantitatively measured using a computer-aided image system (IMS) (ShentengIT Co., Shanghai, China) on digitalized images that were transformed from analogue images taken by a video camera (Panasonic, MV-CP410, Japan). Each field was 72,800 μm², magnification ×200. , ^# compared with the antiscrambled + control siRNA group; ^▲, compared with the anti-382 + control siRNA group. Trx indicates thioredoxin; 3-NT indicates 3-nitrotyrosine. UUO + anti-Sc + CsiRNA indicates UUO mice treated with scrambled anti-miR and control siRNA; UUO + anti-Sc + HsiRNA indicates UUO mice treated with scrambled anti-miR and HSPD1 siRNA; UUO + anti-382 + CsiRNA indicates UUO mice treated with anti-miR-382 and control siRNA; UUO + anti-382 + HsiRNA indicates UUO mice treated with anti-miR-382 and HSPD1 siRNA ().

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