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Anticancer drug 2-methoxyestradiol protects against renal ischemia/reperfusion injury by reducing inflammatory cytokines expression.

Chen YY, Yeh CH, So EC, Sun DP, Wang LY, Hsing CH - Biomed Res Int (2014)

Bottom Line: In vitro, we determined the effect of 2ME2 on reactive oxygen species (ROS) production and cell viability in antimycin-A-treated renal mesangial (RMC) and tubular (NRK52E) cells.Western blotting showed that 2ME2 treatment substantially decreased the expression of activated caspase-9, NF-κB, and HIF-1α but increased the antiapoptotic proteins BCL-2 and BCL-xL in kidney of I/R injury.In vitro, 2MR2 decreased ROS production and increased cell viability in antimycin-A-treated RMC and NRK52E cells. 2ME2 reduces renal I/R injury in mice because it inhibits the expression of ROS and proinflammatory cytokines and induces antiapoptotic proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Biotechnology, National Formosa University, Yunlin County 632, Taiwan.

ABSTRACT

Background: Ischemia/reperfusion (I/R) injury is a major cause of acute renal failure and allograft dysfunction in kidney transplantation. ROS/inflammatory cytokines are involved in I/R injury. 2-Methoxyestradiol (2ME2), an endogenous metabolite of estradiol, inhibits inflammatory cytokine expression and is an antiangiogenic and antitumor agent. We investigated the inhibitory effect of 2ME2 on renal I/R injury and possible molecular actions.

Methods: BALB/c mice were intraperitoneally injected with 2ME2 (10 or 20 mg/kg) or vehicle 12 h before and immediately after renal I/R experiments. The kidney weight, renal function, tubular damages, and apoptotic response were examined 24 h after I/R injury. The expression of mRNA of interleukin-1β, tumor necrosis factor- (TNF) α, caspase-3, hypoxia inducible factor- (HIF) 1α, and proapoptotic Bcl-2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3) in kidney tissue was determined using RT-PCR, while the expression of nuclear factor κB (NF-κB), BCL-2, and BCL-xL, activated caspase-9, and HIF-1α was determined using immunoblotting. In vitro, we determined the effect of 2ME2 on reactive oxygen species (ROS) production and cell viability in antimycin-A-treated renal mesangial (RMC) and tubular (NRK52E) cells.

Results: Serum creatinine and blood urea nitrogen were significantly higher in mice with renal I/R injury than in sham control and in I/R+2ME2-treated mice. Survival in I/R+2ME2-treated mice was higher than in I/R mice. Histological examination showed that 2ME2 attenuated tubular damage in I/R mice, which was associated with lower expression TNF-α, IL-1β, caspase-9, HIF-1α, and BNIP3 mRNA in kidney tissue. Western blotting showed that 2ME2 treatment substantially decreased the expression of activated caspase-9, NF-κB, and HIF-1α but increased the antiapoptotic proteins BCL-2 and BCL-xL in kidney of I/R injury. In vitro, 2MR2 decreased ROS production and increased cell viability in antimycin-A-treated RMC and NRK52E cells.

Conclusions: 2ME2 reduces renal I/R injury in mice because it inhibits the expression of ROS and proinflammatory cytokines and induces antiapoptotic proteins.

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Related in: MedlinePlus

2ME2 reduced expression of HIF-1α, activated caspase-9, and pNF-κB but increased antiapoptotic BCL-xL and BCL-2. Expression levels of HIF-1α (a), pNF-κB (b), BCL-2 (c), BCL-xL (d), and activated caspase-9 (e) in kidneys were determined using western blotting in three groups: sham control: mice undergoing the same procedure without occlusion of the renal pedicle; I/R-only: mice given renal I/R challenge combined with vehicle (normal saline) treatment; I/R+2ME2: I/R-only mice injected with 2ME2 (10 mg/kg, i.p.). β-Actin was the internal control. Representative data are shown. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
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fig5: 2ME2 reduced expression of HIF-1α, activated caspase-9, and pNF-κB but increased antiapoptotic BCL-xL and BCL-2. Expression levels of HIF-1α (a), pNF-κB (b), BCL-2 (c), BCL-xL (d), and activated caspase-9 (e) in kidneys were determined using western blotting in three groups: sham control: mice undergoing the same procedure without occlusion of the renal pedicle; I/R-only: mice given renal I/R challenge combined with vehicle (normal saline) treatment; I/R+2ME2: I/R-only mice injected with 2ME2 (10 mg/kg, i.p.). β-Actin was the internal control. Representative data are shown. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.

Mentions: Western immunoblots of kidney lysates were used to confirm whether 2ME2 protects against apoptosis by inhibiting HIF-1α protein expression. Under baseline conditions, caspase-3 activation was not detectable in the kidneys of any of the mice. Protein levels of HIF-1α (Figure 5(a)), pNF-κB (Figure 5(b)), and activated caspase-9 (Figure 5(e)) were higher in the I/R-only group than in the sham control and I/R+2ME2 group. However, BCL-2 (Figure 5(c)) and BCL-xL (Figure 5(d)) expression were higher in the I/R+2ME2 group than in the I/R-only group.


Anticancer drug 2-methoxyestradiol protects against renal ischemia/reperfusion injury by reducing inflammatory cytokines expression.

Chen YY, Yeh CH, So EC, Sun DP, Wang LY, Hsing CH - Biomed Res Int (2014)

2ME2 reduced expression of HIF-1α, activated caspase-9, and pNF-κB but increased antiapoptotic BCL-xL and BCL-2. Expression levels of HIF-1α (a), pNF-κB (b), BCL-2 (c), BCL-xL (d), and activated caspase-9 (e) in kidneys were determined using western blotting in three groups: sham control: mice undergoing the same procedure without occlusion of the renal pedicle; I/R-only: mice given renal I/R challenge combined with vehicle (normal saline) treatment; I/R+2ME2: I/R-only mice injected with 2ME2 (10 mg/kg, i.p.). β-Actin was the internal control. Representative data are shown. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
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fig5: 2ME2 reduced expression of HIF-1α, activated caspase-9, and pNF-κB but increased antiapoptotic BCL-xL and BCL-2. Expression levels of HIF-1α (a), pNF-κB (b), BCL-2 (c), BCL-xL (d), and activated caspase-9 (e) in kidneys were determined using western blotting in three groups: sham control: mice undergoing the same procedure without occlusion of the renal pedicle; I/R-only: mice given renal I/R challenge combined with vehicle (normal saline) treatment; I/R+2ME2: I/R-only mice injected with 2ME2 (10 mg/kg, i.p.). β-Actin was the internal control. Representative data are shown. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
Mentions: Western immunoblots of kidney lysates were used to confirm whether 2ME2 protects against apoptosis by inhibiting HIF-1α protein expression. Under baseline conditions, caspase-3 activation was not detectable in the kidneys of any of the mice. Protein levels of HIF-1α (Figure 5(a)), pNF-κB (Figure 5(b)), and activated caspase-9 (Figure 5(e)) were higher in the I/R-only group than in the sham control and I/R+2ME2 group. However, BCL-2 (Figure 5(c)) and BCL-xL (Figure 5(d)) expression were higher in the I/R+2ME2 group than in the I/R-only group.

Bottom Line: In vitro, we determined the effect of 2ME2 on reactive oxygen species (ROS) production and cell viability in antimycin-A-treated renal mesangial (RMC) and tubular (NRK52E) cells.Western blotting showed that 2ME2 treatment substantially decreased the expression of activated caspase-9, NF-κB, and HIF-1α but increased the antiapoptotic proteins BCL-2 and BCL-xL in kidney of I/R injury.In vitro, 2MR2 decreased ROS production and increased cell viability in antimycin-A-treated RMC and NRK52E cells. 2ME2 reduces renal I/R injury in mice because it inhibits the expression of ROS and proinflammatory cytokines and induces antiapoptotic proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Biotechnology, National Formosa University, Yunlin County 632, Taiwan.

ABSTRACT

Background: Ischemia/reperfusion (I/R) injury is a major cause of acute renal failure and allograft dysfunction in kidney transplantation. ROS/inflammatory cytokines are involved in I/R injury. 2-Methoxyestradiol (2ME2), an endogenous metabolite of estradiol, inhibits inflammatory cytokine expression and is an antiangiogenic and antitumor agent. We investigated the inhibitory effect of 2ME2 on renal I/R injury and possible molecular actions.

Methods: BALB/c mice were intraperitoneally injected with 2ME2 (10 or 20 mg/kg) or vehicle 12 h before and immediately after renal I/R experiments. The kidney weight, renal function, tubular damages, and apoptotic response were examined 24 h after I/R injury. The expression of mRNA of interleukin-1β, tumor necrosis factor- (TNF) α, caspase-3, hypoxia inducible factor- (HIF) 1α, and proapoptotic Bcl-2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3) in kidney tissue was determined using RT-PCR, while the expression of nuclear factor κB (NF-κB), BCL-2, and BCL-xL, activated caspase-9, and HIF-1α was determined using immunoblotting. In vitro, we determined the effect of 2ME2 on reactive oxygen species (ROS) production and cell viability in antimycin-A-treated renal mesangial (RMC) and tubular (NRK52E) cells.

Results: Serum creatinine and blood urea nitrogen were significantly higher in mice with renal I/R injury than in sham control and in I/R+2ME2-treated mice. Survival in I/R+2ME2-treated mice was higher than in I/R mice. Histological examination showed that 2ME2 attenuated tubular damage in I/R mice, which was associated with lower expression TNF-α, IL-1β, caspase-9, HIF-1α, and BNIP3 mRNA in kidney tissue. Western blotting showed that 2ME2 treatment substantially decreased the expression of activated caspase-9, NF-κB, and HIF-1α but increased the antiapoptotic proteins BCL-2 and BCL-xL in kidney of I/R injury. In vitro, 2MR2 decreased ROS production and increased cell viability in antimycin-A-treated RMC and NRK52E cells.

Conclusions: 2ME2 reduces renal I/R injury in mice because it inhibits the expression of ROS and proinflammatory cytokines and induces antiapoptotic proteins.

Show MeSH
Related in: MedlinePlus