Yang Ding-hua, Zhang Hua, Huang Yu, Zhou Jie
Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 15;89(46):3299-303.
To investigate the effects and mechanisms of methylene chloride (MC) in hepatic ischemic reperfusion injury.
Eighty SD-rats were divided equally into 4 groups: control group (n = 20), donors without any treatment; CoPP group (n = 20), donors injected with CoPP (5 mg/kg, ip) at 24 h; ZnPP group (n = 20), donors injected with ZnPP (20 mg/kg, ip) at 24 h; MC group (n = 20), donors fed with MC (500 mg/kg) per day for 7 days before graft procurement. Syngeneic orthotopic liver transplantation was performed in rats with modified Kamada's two-cuff technique. And SD rats were used as donors (n = 10)and recipients respectively. 5 recipients in each group were sacrificed and the grafts were procured at day 3 after transplantation, the post-operative survival time was observed in the remnant. The tests were determined as following: the level of serum ALT, AST in recipients; heme oxygenase-1 (HO-1) expression of graft was tested by immunohistochemistry and Western blot; the index of graft apoptosis examined by TUNEL method; the pathology of graft assessed by Suzuki's criteria.
The level of serum ALT [(65 +/- 28) U/L], AST [(187 +/- 43) U/L] in CoPP and ALT [(75 +/- 16) U/L], AST [(185 +/- 42) U/L] in MC group was significantly lower than that ALT [(346 +/- 45) U/L], AST [(474 +/- 90) U/L] in control group and ALT [(578 +/- 75) U/L], AST [(1084 +/- 128) U/L] in ZnPP group (P < 0.01). The median expression of HO-1 in MC group was no significantly different with that in control group (P > 0.05). While the median expression of HO-1 in CoPP group was higher than that in control group (P < 0.05). The index of graft apoptosis in MC group and CoPP group, 4.1% +/- 0.6% and 3.2% +/- 0.8% respectively, was significantly lower than that (12.5% +/- 2.4%) in control group and (25.8% +/- 3.1%) in ZnPP group (P < 0.05). Compared with the other two groups, MC and CoPP groups had lesser neutrophil infiltration and a lower grade of hepatocytic injury in grafts. Suzuki's scores in grafts of MC and CoPP groups were lower than that in control and ZnPP groups (P < 0.05). The median post-transplantation survival time of the recipients in MC and CoPP groups was 100 and 93 days respectively while that in control and ZnPP groups was 85 and 12 days (P < 0.05).
Over-expression of HO-1 and MC both have protective effects in hepatic ischemic reperfusion injury.
探讨二氯甲烷（MC）对肝脏缺血再灌注损伤的影响及其机制。
将80只SD大鼠平均分为4组：对照组（n = 20），供体未作任何处理；CoPP组（n = 20），供体于24小时腹腔注射CoPP（5 mg/kg）；ZnPP组（n = 20），供体于24小时腹腔注射ZnPP（20 mg/kg）；MC组（n = 20），在获取移植物前7天，供体每天给予MC（500 mg/kg）。采用改良的Kamada双套管技术对大鼠进行同种异体原位肝移植。分别以SD大鼠作为供体（n = 10）和受体。每组处死5只受体并在移植后第3天获取移植物，观察剩余受体的术后存活时间。检测项目如下：受体血清ALT、AST水平；采用免疫组织化学和蛋白质印迹法检测移植物血红素加氧酶-1（HO-1）表达；采用TUNEL法检测移植物凋亡指数；按照铃木标准评估移植物病理情况。
CoPP组血清ALT水平[（65±28）U/L]、AST水平[（187±43）U/L]以及MC组血清ALT水平[（75±16）U/L]、AST水平[（185±42）U/L]均显著低于对照组血清ALT水平[（346±45）U/L]、AST水平[（474±90）U/L]以及ZnPP组血清ALT水平[（578±75）U/L]、AST水平[（1084±128）U/L]（P < 0.01）。MC组HO-1的中位表达与对照组相比无显著差异（P > 0.05）。而CoPP组HO-1的中位表达高于对照组（P < 0.05）。MC组和CoPP组移植物凋亡指数分别为4.1%±0.6%和3.2%±0.8%，显著低于对照组的（12.5%±2.4%）和ZnPP组的（25.8%±3.1%）（P < 0.05）。与其他两组相比，MC组和CoPP组移植物中性粒细胞浸润较少，肝细胞损伤程度较低。MC组和CoPP组移植物的铃木评分低于对照组和ZnPP组（P < 0.05）。MC组和CoPP组受体移植后的中位存活时间分别为100天和93天，而对照组和ZnPP组分别为85天和12天（P < 0.05）。
HO-1过表达和MC对肝脏缺血再灌注损伤均具有保护作用。
