Activating Nrf-2 Signaling by Sulforaphane Attenuates Unilateral Ureteral Obstruction-Enhanced Apoptosis and Pyroptosis in Kidney
林威佑、戴槐青、鄭劍廷 1 、余宏政
台灣大學醫學院附設醫院 泌尿部1 國立臺灣師範大學生命科學系
Introduction
Clinically, ureteral obstruction is a common problem.Incidence: 3.9% Cause:
○ Ureteral stone○ Benign prostate hyperplasia○ Urothelial tumor○ Other causes such as ureteral stricture
Hegarty et al., 2001
Increased reactive oxygen species (ROS) may promote two types of programmed cell death in the obstructed kidneyapoptosis and pyroptosis
Sulforaphane (SA) via activating Nrf-2 antioxidant signaling may protect UUO-induced renal damage
Method Unilateral ureteral obstruction (UUO)
A well animal model of mimic renal injury leading to interstitial fibrosis
Nagatoya et al., 2002
Left kidney of Female Wistar rats NormalUUOUUO and sulforaphane
Reactive oxygen species (ROS) Fibrosis, ED-1 (macrophage/monocyte)
infiltration, oxidative stress, autophagy, apoptosis and pyroptosis
Immunohistochemisty and Western blot
Result
H.E.Stain
Normal UUO 7day SA UUO 7day
ED1 Stain Macrophage
Normal UUO 7day SA UUO 7day
4HNE Stain Lipid Oxide
Normal UUO 7day SA UUO7day
Beclin Stain Autophagy
SA UUO7day
Normal
UUO 7day
偵測腎臟組織細胞自噬初期蛋白 Beclin-1表現,觀察時間點為 Normal、阻塞七天、阻塞七天 +DL-蘿蔔硫素 (n=3)。*表示與 Normal組有顯著差異 (p<0.05)。
Normal UUO 7day UUO & SA 7day
Beclin 1
control UUO UUO+SA
0.0
0.2
0.4
0.6
0.8
1.0
1.2 *
β-actin
N Engl J Med 2012
ATG5-ATG12 Stain Autophagy
SA UUO7day
Normal
UUO 7day
Atg5-Atg12
偵測腎臟組織細胞自噬過程促自噬泡形成之 Atg5-Atg12蛋白表現,觀察時間點為 Normal、阻塞七天、阻塞七天 + DL-蘿蔔硫素 (n=3)。*表示與 Normal組有顯著差異 (p<0.05)。
Normal UUO 7day UUO & SA 7day
β-actin
control UUO UUO+SA
0.0
0.2
0.4
0.6
0.8
1.0
1.2 *
N Engl J Med 2012
LC3II Stain Autophagy
Normal
UUO 7day
SA UUO7day
LC3II
Normal UUO 7day UUO & SA 7day
偵測腎臟組織細胞自噬之自噬泡形成代表性蛋白 LC3 typeⅡ表現,觀察時間點為 Normal、阻塞七天、阻塞七天 + DL-蘿蔔硫素 (n=3)。*表示與 Normal組有顯著差異 (p<0.05)。
β-actin
control UUO UUO+SA
0.0
0.2
0.4
0.6
0.8
1.0
1.2
*
N Engl J Med 2012
Caspase1 Stain pyropotosis
SA UUO7day
Normal
UUO 7day
IL1-βStain pyropotosis
SA UUO7day
Normal
UUO 7day
Nrf2 Stain
SA UUO7day
Normal
UUO7day
偵測腎臟組織細胞自噬過程 Nrf2蛋白表現,觀察時間點為 Normal、阻塞七天、阻塞七天 + DL-蘿蔔硫素(n=3)。*表示與 control組有顯著差異 (p<0.05)。
Normal UUO 7day UUO & SA 7day
β-actin
Nrf2
control UUO UUO+SA
0.0
0.2
0.4
0.6
0.8
1.0
1.2
*
UOO
Nuclear Nrf-2 translocation
Cytosolic Bax translocation to mitochondria
MOMP
Enhance mitochondrial Cytochrome c release into the cytosol
Cytosolic Nrf-2
sulforaphane
Cytochrome C Stain
Control UUO 7day SA UUO7day
Masson Stain Hydroxylproline
Normal UUO 7day SA UUO7day
Normal UUO7days UUO7&SA days
Hyd
roxylp
rolin
e c
on
ce
nt (m
g/m
l)
0
20
40
60
80
100Normal n=3UUO7daysUUO7days&SA
*
*
Conclusion
Conclusion: Sulforaphane via activating Nrf-2 signaling to preserve mitochondrial function and suppressed UUO-induced renal apoptosis and pyroptosis
Thank you for your attention!!