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SPA0355 attenuates ischemia/reperfusion-induced liver injury in mice.

Bae UJ, Yang JD, Ka SO, Koo JH, Woo SJ, Lee YR, Yu HC, Cho BH, Zhao HY, Ryu JH, Lee SM, Jeon R, Park BH - Exp. Mol. Med. (2014)

Bottom Line: Concomitantly, the expression of NF-κB target genes such as IL-1β, IL-6, TNF-α and iNOS was significantly downregulated.Lastly, the liver antioxidant enzymes superoxide dismutase, catalase and glutathione were upregulated by SPA0355 treatment, which correlated with the reduction in serum malondialdehyde.Our results suggest that SPA0355 pretreatment prior to I/R injury could be an effective method to reduce liver damage.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Chonbuk National University Medical School, Jeonbuk, Republic of Korea.

ABSTRACT
Hepatic ischemia/reperfusion (I/R) injury leads to oxidative stress and acute inflammatory responses that cause liver damage and have a considerable impact on the postoperative outcome. Much research has been performed to develop possible protective techniques. We aimed to investigate the efficacy of SPA0355, a synthetic thiourea analog, in an animal model of hepatic I/R injury. Male C57BL/6 mice underwent normothermic partial liver ischemia for 45 min followed by varying periods of reperfusion. The animals were divided into three groups: sham operated, I/R and SPA0355 pretreated. Pretreatment with SPA0355 protected against hepatic I/R injury, as indicated by the decreased levels of serum aminotransferase and reduced parenchymal necrosis and apoptosis. Liver synthetic function was also restored by SPA0355 as reflected by the prolonged prothrombin time. To gain insight into the mechanism involved in this protection, we measured the activity of nuclear factor-κB (NF-κB), which revealed that SPA0355 suppressed the nuclear translocation and DNA binding of NF-κB subunits. Concomitantly, the expression of NF-κB target genes such as IL-1β, IL-6, TNF-α and iNOS was significantly downregulated. Lastly, the liver antioxidant enzymes superoxide dismutase, catalase and glutathione were upregulated by SPA0355 treatment, which correlated with the reduction in serum malondialdehyde. Our results suggest that SPA0355 pretreatment prior to I/R injury could be an effective method to reduce liver damage.

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Related in: MedlinePlus

Schematic diagram of the experimental protocol. The following assays were performed after the reperfusion: Western blotting and real-time RT–PCR analyses for NF-κB activation (1 h); AST, ALT, ELISA for cytokines and PT assays (6 h); and MPO, MDA, CAT, SOD and GSH assays and western blotting for apoptosis and end-point histology (24 h). ALT, alanine aminotransferase; AST, aspartate aminotransferase; CAT, catalase; MPO, myeloperoxidase; PT, prothrombin time; RT–PCR, real-time RT–PCR; WB, western blotting.
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fig1: Schematic diagram of the experimental protocol. The following assays were performed after the reperfusion: Western blotting and real-time RT–PCR analyses for NF-κB activation (1 h); AST, ALT, ELISA for cytokines and PT assays (6 h); and MPO, MDA, CAT, SOD and GSH assays and western blotting for apoptosis and end-point histology (24 h). ALT, alanine aminotransferase; AST, aspartate aminotransferase; CAT, catalase; MPO, myeloperoxidase; PT, prothrombin time; RT–PCR, real-time RT–PCR; WB, western blotting.

Mentions: Mice were anesthetized with ketamine (100 mgkg−1) and xylazine (10 mgkg−1) by intraperitoneal injection. A midline incision was performed and an atraumatic clip was placed across the portal vein, hepatic artery and bile duct just above the right branch. This interrupted blood flow to the left lateral and median lobes that represents approximately 70% of the total blood supply to the liver. The liver was kept moist with gauze soaked in saline, and body temperature was maintained at 37 °C with a warm blanket throughout ischemia. After 45 min of partial hepatic ischemia, the clip was removed to initiate reperfusion (Figure 1). Sham mice underwent the same operation without vascular occlusion. After the desired time period of reperfusion, the mice were killed by exsanguination under anesthesia and serum samples were collected. Left lateral and median lobes of the liver were collected and immediately fixed in 10% formalin or stored at −80 °C until further analysis.


SPA0355 attenuates ischemia/reperfusion-induced liver injury in mice.

Bae UJ, Yang JD, Ka SO, Koo JH, Woo SJ, Lee YR, Yu HC, Cho BH, Zhao HY, Ryu JH, Lee SM, Jeon R, Park BH - Exp. Mol. Med. (2014)

Schematic diagram of the experimental protocol. The following assays were performed after the reperfusion: Western blotting and real-time RT–PCR analyses for NF-κB activation (1 h); AST, ALT, ELISA for cytokines and PT assays (6 h); and MPO, MDA, CAT, SOD and GSH assays and western blotting for apoptosis and end-point histology (24 h). ALT, alanine aminotransferase; AST, aspartate aminotransferase; CAT, catalase; MPO, myeloperoxidase; PT, prothrombin time; RT–PCR, real-time RT–PCR; WB, western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150932&req=5

fig1: Schematic diagram of the experimental protocol. The following assays were performed after the reperfusion: Western blotting and real-time RT–PCR analyses for NF-κB activation (1 h); AST, ALT, ELISA for cytokines and PT assays (6 h); and MPO, MDA, CAT, SOD and GSH assays and western blotting for apoptosis and end-point histology (24 h). ALT, alanine aminotransferase; AST, aspartate aminotransferase; CAT, catalase; MPO, myeloperoxidase; PT, prothrombin time; RT–PCR, real-time RT–PCR; WB, western blotting.
Mentions: Mice were anesthetized with ketamine (100 mgkg−1) and xylazine (10 mgkg−1) by intraperitoneal injection. A midline incision was performed and an atraumatic clip was placed across the portal vein, hepatic artery and bile duct just above the right branch. This interrupted blood flow to the left lateral and median lobes that represents approximately 70% of the total blood supply to the liver. The liver was kept moist with gauze soaked in saline, and body temperature was maintained at 37 °C with a warm blanket throughout ischemia. After 45 min of partial hepatic ischemia, the clip was removed to initiate reperfusion (Figure 1). Sham mice underwent the same operation without vascular occlusion. After the desired time period of reperfusion, the mice were killed by exsanguination under anesthesia and serum samples were collected. Left lateral and median lobes of the liver were collected and immediately fixed in 10% formalin or stored at −80 °C until further analysis.

Bottom Line: Concomitantly, the expression of NF-κB target genes such as IL-1β, IL-6, TNF-α and iNOS was significantly downregulated.Lastly, the liver antioxidant enzymes superoxide dismutase, catalase and glutathione were upregulated by SPA0355 treatment, which correlated with the reduction in serum malondialdehyde.Our results suggest that SPA0355 pretreatment prior to I/R injury could be an effective method to reduce liver damage.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Chonbuk National University Medical School, Jeonbuk, Republic of Korea.

ABSTRACT
Hepatic ischemia/reperfusion (I/R) injury leads to oxidative stress and acute inflammatory responses that cause liver damage and have a considerable impact on the postoperative outcome. Much research has been performed to develop possible protective techniques. We aimed to investigate the efficacy of SPA0355, a synthetic thiourea analog, in an animal model of hepatic I/R injury. Male C57BL/6 mice underwent normothermic partial liver ischemia for 45 min followed by varying periods of reperfusion. The animals were divided into three groups: sham operated, I/R and SPA0355 pretreated. Pretreatment with SPA0355 protected against hepatic I/R injury, as indicated by the decreased levels of serum aminotransferase and reduced parenchymal necrosis and apoptosis. Liver synthetic function was also restored by SPA0355 as reflected by the prolonged prothrombin time. To gain insight into the mechanism involved in this protection, we measured the activity of nuclear factor-κB (NF-κB), which revealed that SPA0355 suppressed the nuclear translocation and DNA binding of NF-κB subunits. Concomitantly, the expression of NF-κB target genes such as IL-1β, IL-6, TNF-α and iNOS was significantly downregulated. Lastly, the liver antioxidant enzymes superoxide dismutase, catalase and glutathione were upregulated by SPA0355 treatment, which correlated with the reduction in serum malondialdehyde. Our results suggest that SPA0355 pretreatment prior to I/R injury could be an effective method to reduce liver damage.

Show MeSH
Related in: MedlinePlus