苦参碱对大鼠肝脏缺血/再灌注损伤后肝功能和TNF-α的影响

Influence of matrine on liver functions and TNF-α level following liver ischemic reperfusion injury in rats

目的探讨苦参碱预处理对大鼠缺血/再灌注肝损伤(HIRI)的保护作用及其机制。方法健康成年雄性SD大鼠75只,随机分为5组,每组15只,15只作为空白对照组(SO组),不阻断肝脏血流;60只以夹闭门静脉和肝动脉缺血70 min后进行再灌注方法建立HIRI模型,然后再分为4组,分别为模型组即缺血/再灌注+生理盐水组(NS组)、缺血/再灌注+苦参碱组(MT组),MT组依剂量不同分别为MT1、MT2、MT3组(苦参碱剂量分别为7.5、15.0、30.0 mg/kg),MT组在夹闭门静脉和肝动脉前经门静脉主干注入苦参碱。NS组注入生理盐水,阻断供血70 min后恢复灌注,在恢复灌注2 h后采集相关标本待测。检测各组大鼠肝功能指标,包括血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)及肝组织匀浆中TNF-α的含量,同时观察肝组织病理形态学改变。结果病理组织学检查显示MT组肝细胞损伤程度较模型组轻,且MT3组最轻;模型组肝功能指标明显高于SO组及MT各组(P均<0.01);MT各组中,MT1组明显高于MT3组(P<0.05或P<0.01),即苦参碱与肝功能指标下降存在剂量效应关系。SO组肝组织中TNF-α含量较少为(54.76±11.32)pg/mg·prot,模型组TNF-α含量最高为(2 175.67±131.64)pg/mg·prot,MT1含量为(1 664.47±109.21)pg/mg·prot,MT2含量为(1 561.52±104.52)pg/mg·prot,MT3含量为(1 245.17±96.69)pg/mg·prot,MT各组随剂量增加TNF-α含量降低,3组与模型组比较差异均有统计学意义(P均<0.05),MT3组低于MT1、MT2(P均<0.05)。结论苦参碱对缺血/再灌注中肝细胞损伤有保护作用,且存在剂量相关性,其机制可能与苦参碱抑制TNF-α释放有关。

Objective To explore the protective effects of matrine pre-treatment on the hepatic ischemia reperfusion injury（ HIRI） in rats and its potential mechanism. Methods Seventy-five healthy male SD rats were randomly divided into five groups： 15 rats were served as blank control group（ SO group）,and the hepatic blood flow was not be blocked in this group; in the remaining 60 rats,HIRI model was established by clip-closed portal vein and hepatic artery followed by reperfusion,and then the rats were divided into four groups（ n = 15 each） ： model group（ NS group）,MT group including MT1,MT2 and MT3 subgroups with different doses of matrine（ 7. 5,15. 0,30. 0 mg / kg）. In MT subgroups,different doses of matrine were respectively injected through main portal vein before closing portal vein and hepatic artery,while normal saline was injected in the same method in NS group. The blood perfusion was recovered 70 minutes after blocking blood supply,and the related samples were collected 2 hours after recovering perfusion to be measured. In all groups,the indexes of liver function including serum alanine aminotransferase（ ALT）,aspartate aminotransferas（ AST）,lactate dehydrogenase（ LDH）and tumor necrosis factor（ TNF）-α content in homogenate of liver tissue were detected and the morphopathology changes of liver were observed. Results Morphopathology examination showed that the injury degree of liver cells in MT group was lighter than that in NS group,and it was the lightest in MT3 group. The indexes of liver functions in NS group were significantly higher than those in SO group and all MT subgroups（ all P〈0. 01）,and the indexes of liver functions in MT1 subgroup were significantly higher than those in MT3 subgroup（ P〈0. 05 or P〈0. 01）,namely,matrine was associated with descending of liver functions indexes in a dose-dependent manner. The TNF-α content in liver tissues in SO group [（ 54. 76± 11. 32） pg / mg prot] was lower,and it in NS group[（ 2175. 67 ± 131. 64） pg / mg prot] was the lowest. The TNF-α contents in MT subgroups decreased with the increase of matrine doses,and there were significant differences in the TNF-αcontents（ all P〈0. 05） between MT subgroups and NS group. The TNF-α contents in MT3 subgroup decreased compared with MT1 and MT2 subgroups（ all P〈0. 05）. Conclusion Matrine has a protective effect against HIRI,and the effect exists correlation with matrine doses,and its mechanism may be associated with inhibiting the delivery of TNF-α.