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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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Effect of 2ME2 on renal morphology in renal I/R injury. Renal injury was evidenced by massive TEC necrosis or the damage or loss of brush border (arrows) and by tubular dilation and cast formation (arrowheads). (a) Sham control; (b) I/R-only; (c) I/R + 2ME2: I/R mice injected with 2ME2 (10 mg/kg, i.p.); and (d) I/R+2ME2: I/R mice injected with 2ME2 (20 mg/kg, i.p.). H&E staining was used to evaluate the degree of acute renal tubules damage in renal I/R injury. Representative data are shown. Sections were observed under a microscope (×400). Scale bar is 50 μm. The histological scores of tubular injury in the cortex (e), corticomedullary junction (CMJ) (f), and medulla (g) of all groups of mice are shown. Values are mean ± SD. 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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fig1: Effect of 2ME2 on renal morphology in renal I/R injury. Renal injury was evidenced by massive TEC necrosis or the damage or loss of brush border (arrows) and by tubular dilation and cast formation (arrowheads). (a) Sham control; (b) I/R-only; (c) I/R + 2ME2: I/R mice injected with 2ME2 (10 mg/kg, i.p.); and (d) I/R+2ME2: I/R mice injected with 2ME2 (20 mg/kg, i.p.). H&E staining was used to evaluate the degree of acute renal tubules damage in renal I/R injury. Representative data are shown. Sections were observed under a microscope (×400). Scale bar is 50 μm. The histological scores of tubular injury in the cortex (e), corticomedullary junction (CMJ) (f), and medulla (g) of all groups of mice are shown. Values are mean ± SD. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.

Mentions: To examine the effects of 2ME2 on renal I/R injury, the mice were injected with vehicle or 2ME2 i.p. as described in Materials and Methods. Twenty-four hours after I/R, renal function was assessed based on serum creatinine and BUN levels. The kidney weight and the serum BUN and creatinine concentrations were significantly increasing in I/R-only mice. In I/R+2ME2-treated mice, either in 10 or 20 mg/kg groups, the increase was reduced (Table 1). Histological analysis of the kidney sections showed significant renal tubular injury in I/R group (Figure 1). Kidney I/R-induced tubular cell injuries were characterized by vacuolization, loss of brush borders, sloughing of tubular cells into the lumen, and flattening of the tubular epithelium (Figure 1(b)), which were less frequently seen in the I/R+2ME2 (10 mg/kg) (Figure 1(c)) and I/R+2ME2 (20 mg/kg) groups (Figure 1(d)). The severity of renal I/R injury was also assessed by measuring the histological injury score. The I/R group had higher histological scores than did the sham control group for the cortex, corticomedullary junction (CMJ), and medulla (Figures 1(e), 1(f), and 1(g), resp.). Both the I/R+2ME2 (10 mg/kg) and I/R+2ME2 (20 mg/kg) groups had significantly lower histological scores than did the I/R-only group (P < 0.05). Mice survival rates 7 days after there had been renal I/R injury were 67, 83, and 100% for the I/R+vehicle, I/R+2ME2 (10 mg/kg), and I/R+2ME2 (20 mg/kg) groups, respectively (Figure 2).


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)

Effect of 2ME2 on renal morphology in renal I/R injury. Renal injury was evidenced by massive TEC necrosis or the damage or loss of brush border (arrows) and by tubular dilation and cast formation (arrowheads). (a) Sham control; (b) I/R-only; (c) I/R + 2ME2: I/R mice injected with 2ME2 (10 mg/kg, i.p.); and (d) I/R+2ME2: I/R mice injected with 2ME2 (20 mg/kg, i.p.). H&E staining was used to evaluate the degree of acute renal tubules damage in renal I/R injury. Representative data are shown. Sections were observed under a microscope (×400). Scale bar is 50 μm. The histological scores of tubular injury in the cortex (e), corticomedullary junction (CMJ) (f), and medulla (g) of all groups of mice are shown. Values are mean ± SD. 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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fig1: Effect of 2ME2 on renal morphology in renal I/R injury. Renal injury was evidenced by massive TEC necrosis or the damage or loss of brush border (arrows) and by tubular dilation and cast formation (arrowheads). (a) Sham control; (b) I/R-only; (c) I/R + 2ME2: I/R mice injected with 2ME2 (10 mg/kg, i.p.); and (d) I/R+2ME2: I/R mice injected with 2ME2 (20 mg/kg, i.p.). H&E staining was used to evaluate the degree of acute renal tubules damage in renal I/R injury. Representative data are shown. Sections were observed under a microscope (×400). Scale bar is 50 μm. The histological scores of tubular injury in the cortex (e), corticomedullary junction (CMJ) (f), and medulla (g) of all groups of mice are shown. Values are mean ± SD. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
Mentions: To examine the effects of 2ME2 on renal I/R injury, the mice were injected with vehicle or 2ME2 i.p. as described in Materials and Methods. Twenty-four hours after I/R, renal function was assessed based on serum creatinine and BUN levels. The kidney weight and the serum BUN and creatinine concentrations were significantly increasing in I/R-only mice. In I/R+2ME2-treated mice, either in 10 or 20 mg/kg groups, the increase was reduced (Table 1). Histological analysis of the kidney sections showed significant renal tubular injury in I/R group (Figure 1). Kidney I/R-induced tubular cell injuries were characterized by vacuolization, loss of brush borders, sloughing of tubular cells into the lumen, and flattening of the tubular epithelium (Figure 1(b)), which were less frequently seen in the I/R+2ME2 (10 mg/kg) (Figure 1(c)) and I/R+2ME2 (20 mg/kg) groups (Figure 1(d)). The severity of renal I/R injury was also assessed by measuring the histological injury score. The I/R group had higher histological scores than did the sham control group for the cortex, corticomedullary junction (CMJ), and medulla (Figures 1(e), 1(f), and 1(g), resp.). Both the I/R+2ME2 (10 mg/kg) and I/R+2ME2 (20 mg/kg) groups had significantly lower histological scores than did the I/R-only group (P < 0.05). Mice survival rates 7 days after there had been renal I/R injury were 67, 83, and 100% for the I/R+vehicle, I/R+2ME2 (10 mg/kg), and I/R+2ME2 (20 mg/kg) groups, respectively (Figure 2).

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