| Objective:This study aimed to establish the rat model of warm partial hepatic ischemia- reperfusion, and investigate the protective and anti-inflammatory effects of isoflurane on warm hepatic ischemia-reperfusion injury (IRI) in rats.Materials and Methods:Thirty-two female Sprague-Dawley rats were divided equally into 4 groups (n=8): PB-Sham group in which the rats were anesthetized by intraperitoneal injection of pentobarbital sodium (1.0%, 40mg﹒kg-1, PB) and received a sham operation without occlusion of liver blood flow; PB-IR group whose rats underwent partial hepatic ischemia-reperfusion (IR) after anesthesia; Iso-Sham group in which inhalation of 1.0 MAC isoflurane and sham operation was performed; Iso-IR group in which 1.0 MAC isoflurane was inhaled for 4 h and IR was performed. rat model of warm partial hepatic ischemia-reperfusion was established by clamping the hepatic arteries and hilar vessels distributing to the left and median lobes to induce partial hepatic ischemia (70%) for 60 min followed by reperfusion for 3 h. The rats were killed 3 h after declamping, and specimens of liver tissue and blood were obtained. The serum ALT and AST were detected as liver damage markers. Viability of myeloperoxidase(MPO) in liver were measured. The expressions of ICAM-1 and TNF-αwere detected by RT-PCR; The protein level of ICAM-1 in the liver was detected by immunohistochemistry and Western blotting. The histological observation was carred out with hematoxylin and eosin staining.Results:1. Changes of serum ALT and AST The serum levels of AST and ALT in PB-IR group were significantly higher than those in other 3 groups (P<0.05). Serum enzyme in Iso-IR group was increased compared with PB-Sham group or Iso-Sham group, but decreased with PB-IR group (P<0.05).2. Liver mRNA changes of ICAM-1 and TNF-αmRNA expressions of ICAM-1 and TNF-αin PB-IR groups were significantly higher than those in other 3 groups (P<0.05). Isoflurane attenuated the mRNA expressions of ICAM-1 and TNF-α(P<0.05).3. Immunohistochemical assay of liver ICAM-1 Hepatic sinusoidal endothelial cells (SECs), hepatocytes, central veins, and interlobular veins showed weak staining of ICAM-1 in PB-Sham group and Iso-Sham group. In the PB-IR group, strong staining to ICAM-1 was observed on SECs, central veins and interlobular veins, and weak staining on hepatocytes. But in Iso-IR group the positive area and intensity were much lower (P<0.05).4. Results of Western blotting The expression level of ICAM-1 was analyzed with integral optical density of the blots. In the PB-IR group, ICAM-1 expression was increased in comparison to other 3 groups (P<0.05). The expression was significantly higher in Iso-IR group than in PB-Sham group and Iso-Sham group(P<0.05).5. MPO level The MPO in Iso-IR group was significantly higher than PB-Sham group and Iso-Sham group(P<0.05), but significantly lower than PB-IR group (P<0.05), MPO in PB-IR group were significantly highest (P<0.05).6. Morphological changes of liver The hepatic tissue was normal both in PB-Sham group and Iso-Sham group. In the PB-IR group, there were obviously localized hepatocytic necrosis, even coagulation necrosis. Some obvious fatty degeneration of hepatocyte were seen around lobular peripheral and central veins. The structure of hepatic cords was destroyed. In Iso-IR group, there were congestion and stretch both in central veins and sinudoids. Hepatocytic degeneration, and even spotty necrosis and fatty degeneration were observed. The neutrophils gathering were seen around the portal region. The basic structure of hepatic cords was reserved.Conclusion:1. Rats treated with 1.0 MAC isoflurane during warm partial(70%) hepatic ischemia 60 min and 3 h of reperfusion have significantly lower serum ALT and AST compared with rats anesthetized with pentobarbital sodium subjected to hepatic ischemia–reperfusion injury. 2. Gene and protein expression of ICAM-1 in rat hepatic tissue are significantly increased by hepatic ischemia–reperfusion injury after pentobarbital sodium anesthesia.3. Isoflurane can significantly inhibit gene and protein expression of ICAM-1 in rat hepatic ischemia–reperfusion injury in comparison to rats with pentobarbital sodium anesthesia subjected to hepatic ischemia–reperfusion injury.4. mRNA expression of TNF-αin rat liver is significantly increased by hepatic ischemia–reperfusion injury after pentobarbital sodium anesthesia. Isoflurane can significantly inhibit expression of TNF-αin rat hepatic ischemia–reperfusion injury.5. Viability of MPO in the liver is significantly increased by hepatic ischemia–reperfusion injury after pentobarbital sodium anesthesia; Isoflurane can significantly inhibit MPO alteration in rat liver ischemia–reperfusion injury compared with rats anesthetized with pentobarbital sodium subjected to hepatic ischemia–reperfusion injury.
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