Limits...
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.

Show MeSH

Related in: MedlinePlus

TUNEL staining of kidneys. Apoptotic cells in the kidneys of each group of mice were determined using TUNEL staining. All groups: n = 6. (a) There is no fluorescent intensity in the control kidney with saline treatment. (b) Apoptotic cells displaying high fluorescence intensity were detected in sections from I/R-only kidneys; however, (c) the green fluorescence of apoptotic nuclei is rare in tubular epithelial cells of the kidneys with 2ME2 (20 mg/kg) treatment. Sections were observed under a fluorescence microscope (×400). Scale bar is 50 μm. Representative data are shown. (d) Apoptotic cell counts in kidneys are averages of 10 high power fields (HPFs) from each group of mice. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4151070&req=5

fig4: TUNEL staining of kidneys. Apoptotic cells in the kidneys of each group of mice were determined using TUNEL staining. All groups: n = 6. (a) There is no fluorescent intensity in the control kidney with saline treatment. (b) Apoptotic cells displaying high fluorescence intensity were detected in sections from I/R-only kidneys; however, (c) the green fluorescence of apoptotic nuclei is rare in tubular epithelial cells of the kidneys with 2ME2 (20 mg/kg) treatment. Sections were observed under a fluorescence microscope (×400). Scale bar is 50 μm. Representative data are shown. (d) Apoptotic cell counts in kidneys are averages of 10 high power fields (HPFs) from each group of mice. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.

Mentions: We next determined whether 2ME2 reduces apoptosis in renal I/R injury. Representative kidney histological sections from sham control, I/R-only, and I/R+2ME2-treated mice were stained using TUNEL assay to detect apoptotic cells. Apoptosis was more prominent in the kidney sections from I/R-only mice than in those from sham control mice (Figure 4). A large number of apoptotic nuclei in renal tubular epithelial cells were detected in I/R-only kidneys (Figure 4(b)). Compared with I/R-only mice, markedly fewer apoptotic nuclei in kidney were detected in I/R+2ME2-treated mice (Figures 4(c) and 4(d)).


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)

TUNEL staining of kidneys. Apoptotic cells in the kidneys of each group of mice were determined using TUNEL staining. All groups: n = 6. (a) There is no fluorescent intensity in the control kidney with saline treatment. (b) Apoptotic cells displaying high fluorescence intensity were detected in sections from I/R-only kidneys; however, (c) the green fluorescence of apoptotic nuclei is rare in tubular epithelial cells of the kidneys with 2ME2 (20 mg/kg) treatment. Sections were observed under a fluorescence microscope (×400). Scale bar is 50 μm. Representative data are shown. (d) Apoptotic cell counts in kidneys are averages of 10 high power fields (HPFs) from each group of mice. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4151070&req=5

fig4: TUNEL staining of kidneys. Apoptotic cells in the kidneys of each group of mice were determined using TUNEL staining. All groups: n = 6. (a) There is no fluorescent intensity in the control kidney with saline treatment. (b) Apoptotic cells displaying high fluorescence intensity were detected in sections from I/R-only kidneys; however, (c) the green fluorescence of apoptotic nuclei is rare in tubular epithelial cells of the kidneys with 2ME2 (20 mg/kg) treatment. Sections were observed under a fluorescence microscope (×400). Scale bar is 50 μm. Representative data are shown. (d) Apoptotic cell counts in kidneys are averages of 10 high power fields (HPFs) from each group of mice. All groups: n = 6. *P < 0.05 compared with control mice and #P < 0.05 compared with I/R-only mice.
Mentions: We next determined whether 2ME2 reduces apoptosis in renal I/R injury. Representative kidney histological sections from sham control, I/R-only, and I/R+2ME2-treated mice were stained using TUNEL assay to detect apoptotic cells. Apoptosis was more prominent in the kidney sections from I/R-only mice than in those from sham control mice (Figure 4). A large number of apoptotic nuclei in renal tubular epithelial cells were detected in I/R-only kidneys (Figure 4(b)). Compared with I/R-only mice, markedly fewer apoptotic nuclei in kidney were detected in I/R+2ME2-treated mice (Figures 4(c) and 4(d)).

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