淫羊藿素脂质体抗大鼠肝脏缺血再灌注损伤

目的 探讨淫羊藿素脂质体对大鼠肝脏缺血再灌注（I/R）损伤的作用及其机制。方法 建立大鼠70%肝脏I/R损伤模型。将120只雄性大鼠随机分成淫羊藿素脂质体+I/R损伤（ICT+I/R）组、空白脂质体+I/R损伤（LIP+I/R）组、单纯I/R损伤组和假手术（Sham）组，每组再随机分成2 h和6 h两个亚组。Sham组仅游离肝门，其余各组均行70%肝脏缺血60 min。血流阻断前10 min，ICT+I/R组、LIP+I/R组分别经门静脉注射淫羊藿素脂质体（1.5 mg/kg）、空白脂质体（与淫羊藿素脂质体等体积），I/R组和Sham组不行任何预处理。经2、6 h血流再灌注后，采集各组血液及肝脏组织标本，检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）水平以及肝组织中超氧化物歧化酶（SOD）、丙二醛（MDA）、一氧化氮（NO）、一氧化氮合酶（NOS）、诱导型一氧化氮合酶（iNOS）、内皮型一氧化氮合酶（eNOS）以及髓过氧化物酶（MPO）含量。采用H-E染色法观察肝脏组织形态，TUNEL染色法观察肝细胞凋亡情况并测定凋亡指数（AI）。结果 再灌注2 h，ICT+I/R组的ALT、MDA、MPO、AI 较LIP+I/R组和I/R组下降，差异有统计学意义（P<0.01）；再灌注6 h，与LIP+I/R组和I/R组相比，ICT+I/R组的ALT、AST、MDA、AI下降，同时SOD、NO、NOS、eNOS升高，差异有统计学意义（P<0.05，P<0.01）。结论 淫羊藿素脂质体能够通过增加SOD含量、减少MDA的生成，促进eNOS表达的升高、增加NO的含量以及抑制MPO的聚集、减少肝细胞凋亡等多途径发挥抗大鼠肝脏I/R损伤的保护作用。
Objective To investigate the protective effect of icaritin liposme against hepatic ischemia /reperfusion (I/R) injury in rats and its mechanisms. Methods A total of 120 male SD rats were randomly divided into four groups: icaritin liposome+I/R (ICT+I/R) group, vechicle liposome+I/R (LIP+I/R) group, I/R group and sham operation (Sham) group. Each group was randomly divided into two subgroups of 2 h and 6 h. The rats in the Sham group were only with free hilum, and the other three groups were subjected to 70% liver ischemia for 60 min. Icaritin liposome (1.5 mg/kg) or vechile lipsome (with same volume of icaritin liposome) were intraportal venously injected in the ICT+I/R and LIP+I/R groups at 10 min before ischemia, without any pretreatment in the I/R group. Blood and liver tissue samples in each group were obtained at 2 h and 6 h after reperfusion to measure the levels of serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST), and the contents of superoxide dismutase (SOD), malondialdehyd (MDA), nitric oxide (NO), nitric oxide synthase (NOS), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and myeloperoxidase (MPO) in liver tissues. The morphology of liver tissues was observed by H-E staining. The apoptosis of liver cells and apoptosis index (AI) were calculated by TUNEL staining. Results Compared with the LIP+I/R and IR groups, ALT level, MDA and MPO contents, and AI were significantly decreased in the ICT+I/R group at 2 h after reperfusion (P<0.05, P<0.01). At 6 h after reperfusion, the ALT and AST levels, MDA content, and AI in the ICT+I/R group were significantly reduced, and the contents of SOD, NO, NOS, and eNOS were significantly increased compared with the LIP+I/R and IR groups (P<0.05, P<0.01). Conclusion Icaritin liposome can reduce liver I/R injury