| Objective:To study the surgery techniques in rat orthotopic liver transplantation and the prophylaxis and management of perioperative complications.Methods:The rat orthotopic liver transplantation model was established by Kamada's two cuff techniques with some modification.Sprague-Dawley(SD) rats were used as donors and recipients.Results:160 cases of rat orthotopic liver transplantation operations were performed in total.120 cases were performed in the early phage,and the 48hours survival was about 46.7%.40 cases were done in the advantage phage,and 48hours survival was about 95.0%.Conclusions:The sophisticated microsurgical techniques and the delicate surgical manipulation is the key to successful establishment of rat orthotopic liver transplantation model. Objectives:To study the the effects and mechanisms of riO-1 system in IRI,providing an experimental evidence to prevent IRI in clinical organ transplantation.Methods:Established rat orthotopic liver transplantation with modified Kamada's two cuff technique.SD rats were used as donors and recipients respectively.40 rats were separated into 4 groups in random,five pairs in each group.Group 1 was a control group without any treatments,donors in group 2 were pretreated with CoPP(5mg/kg,intraperitoneally)at-24 hr.donors in group 3 were pretreated with ZnPP(20mg/kg,intraperitoneally)at-24h.donors in group 4 were treated with methylene chloride(MC 500 mg/kg,orally)once a day for seven days before graft havesting.Items were tested as following:(1)The level of donors serum ALT,AST and COHb/Hb%before the graft harvesting. (2)The level of serum ALT,AST in graft after transplantation at day 3.(3)The level of HO-1 expression of the graft after transplantation was tested by immunohistology and Western blot at day 3.(4)The apoptosis of grafts after transplantation was examined by TUNEL method at the tday 3.(5)The pathology changes on ischemia reperfusion injury of graft was evaluated by International Suzuki's Criteria.Results:(1)Compared to other three groups,the level of COHb/Hb%in MC group was significantly increased(P＜0.05).(2)The level of serum ALT and AST in CoPP and MC group was significantly lower than that in the other two groups(P＜0.05).(3)The expression of H0-1 in MC group was no significant different with that in control group(P＞0.05).while the expression of H0-1 in CoPP group was higher than that in control group(P＜0.01).(4)Compared to control group,the index of apoptosis of grafts was decreased in MC group and CoPP group(P＜0.01).(5)Compared to the other two groups,MC group and CoPP group had fewer neutrophil infiltration and lower grade of liver cell injury. Suzuki's scores of MC(0.76±0.52)and COPP(0.76±0.59)group were lower than control group(1.32±0.74)and ZnPP group(1.80±0.70)(P＜0.05).Conclusions:1.Over expression of HO-1 has a protect effect in liver ischemia reperfusion injury.2.As to HO-1,Methylene chloride also has a protect effect in liver ischemia reperfusion injure,that maybe induced by its product CO via its anti-inflammatory and anti-apoptotic functions.3.As a Carbon Monoxide producer,Methylene chloride is safe and effective,maybe has a good prospect in clincal... |
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