摘要目的 探讨雷公藤甲素(TPT)预处理减轻小鼠肝脏缺血再灌注损伤的作用及其作用机制.方法 将60只雄性C57BL/6小鼠分成4组,每组15只.(1)TPT手术组:手术方法参照Kobayashi法,术中夹闭肝门静脉左支、肝动脉左支及左肝管,90min后开放血管,建立小鼠肝脏缺血再灌注损伤模型;(2)TPT假手术组:手术方式同TPT手术组,但术中不阻断血流,仅用生理盐水(NS)纱布覆盖切口90 min;(3)NS手术组:手术方式同TPT手术组;(4)NS假手术组:手术方式同TPT假手术组.两TPT组小鼠于术前1周开始腹腔注射TPT0.1 mg·kg-1·d-1,术前1 h加用1次,而两NS组同期仅腹腔注射等体积无菌NS.再灌注后24 h,检测各组小鼠肝功能和观察肝组织病理学变化,采用流式细胞术检测各组TH17细胞占单个核细胞的比例,采用实时聚合酶链反应(PCR)检测各组肝组织中IL-17和ROR-γt mRNA的表达,采用酶联免疫吸附试验(ELISA)检测血清中IL-6、IL-17和TGF-β的含量.结果 TPT手术组肝功能的损伤较NS手术组明显减轻(P＜0.05);NS假手术组、TPT假手术组、NS手术组和TPT手术组TH17细胞占单个核细胞的比例分别为(0.72±00.23)%、(0.41±0.18)%、(4.26±0.82)%和(1.77±0.53)%;两TPT组IL-17和ROR-γt mRNA的表达量以及外周血中IL-6、IL-17和TGF-β的含量,均明显低于相应的NS组(P＜0.05).结论 低剂量TPT预处理能够减轻小鼠肝脏缺血再灌注损伤,这可能与TPT预处理抑制小鼠体内Th17细胞分化、发育及其功能有关.

abstractsObjective To investigate the protective effect of triptolide (TPT) pretreatment against liver ischemia/reperfusion (I/R) injury in mice and the possible mechanism. Methods Sixty male C57BL/6 mice were randomly divided into four groups (15/group): ( 1 ) TPT I/R group: The mouse partial liver model of I/R injury was established by the method of Koba-yashi. The portal triad (hepatic artery, portal vein, and bile duct) was occluded with a microvascular clamp for 90 min and 24 h reperfusion; (2) Sham group with TPT: Mice underwent surgical procedures including isolation of the portal triad without occlusion; (3) Saline I/R group: Surgery was performed as the same in the TPT I/R group, (4): Sham group with saline: Surgery was performed as the same in the TPT sham group, and the mice were pretreated with either saline or TPT (0. 1 mg · kg-1 day-1 ) by intraperitoneal injection for one week. The samples were collected at the 24th h after the I/R injury.The serum ALT and AST levels were determined, the histologic changes were observed by HE staining, the percentage of Th17 cells among mononuclear cells in liver tissue was analyzed by flow cytometry, the expression of IL-17 and ROR-γt mRNA was detected by real-time PCR, and the serum IL-6, IL-17 and TGF-β levels were measured by enzyme-linked immunosorbent assay (ELISA).Results Serum ALT and AST levels were significantly decreased and the histological damage was significantly alleviated in the TPT I/R group as compared with saline I/R group (P＜0. 05). The percentage of Th17 cells among mononuclear cells in TPT I/R group, TPT sham group, saline I/R group, TPT saline group was ( 1.77 ± 0. 53)％, (0. 41± 0. 18)％, (4. 26 ± 0. 82)％ and (0. 72 ± 0. 23) ％ in liver tissue, respectively. The expression levels of the IL-17 and ROR-γt mRNA in the liver tissue, and IL-6, IL-17 and TGF-β levels in the serum were significantly lower in TPT I/R group than in saline I/R group (P＜0. 05). Conclusion Pretreatment with low dose of TPT could effectively protect the liver from I/R injury in mice, which may be related to the inhibition of Th17 cells.