title: Anticancer drug 2-methoxyestradiol protects against renal ischemia/reperfusion injury by reducing inflammatory cytokines expression.
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.
authors.affiliation: Department of Medical Research, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Biotechnology, National Formosa University, Yunlin County 632, Taiwan.
authors.forename: Ying-Yin
authors.initials: YY
authors.lastname: Chen
authors.affiliation: Department of Medicinal Botanicals and Health Applications, Da-Yeh University, Changhua 515, Taiwan.
authors.forename: Ching-Hua
authors.initials: CH
authors.lastname: Yeh
authors.affiliation: Department of Anesthesia & Medical Research, China Medical University, An Nan Hospital, Tainan 710, Taiwan.
authors.forename: Edmund Cheung
authors.initials: EC
authors.lastname: So
authors.affiliation: Department of Surgery, Chi-Mei Medical Center, Tainan 710, Taiwan.
authors.forename: Ding-Ping
authors.initials: DP
authors.lastname: Sun
authors.affiliation: Department of Medical Research, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Anaesthesiology, Chi-Mei Medical Center, Tainan 710, Taiwan.
authors.forename: Li-Yun
authors.initials: LY
authors.lastname: Wang
authors.affiliation: Department of Medical Research, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Anaesthesiology, Chi-Mei Medical Center, Tainan 710, Taiwan ; Department of Anesthesiology, Taipei Medical University, Taipei 110, Taiwan.
authors.forename: Chung-Hsi
authors.identifier: 0000-0002-6091-5371
authors.initials: CH
authors.lastname: Hsing
mesh_terms: D000077584:2-Methoxyestradiol; D058186:Acute Kidney Injury / Q000235:genetics / Q000473:pathology; D000818:Animals; D016207:Cytokines / Q000096:biosynthesis*; D004958:Estradiol / Q000008:administration & dosage / Q000031:analogs & derivatives*; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000473:pathology; D016030:Kidney Transplantation / Q000009:adverse effects; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis; D015427:Reperfusion Injury / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology
chemical_list: D016207:Cytokines; D012333:RNA, Messenger; D004958:Estradiol; D000077584:2-Methoxyestradiol