硫氧还蛋白相互作用蛋白在大鼠急性肾损伤中的作用

Role of thioredoxin-interacting protein in rat model of acute renal injury

摘要:

目的 探讨硫氧还蛋白相互作用蛋白（TXNIP）在大鼠急性肾损伤中的作用。方法 16 只雄性Sprague Dawley 大鼠随机分为假手术组（S），缺血再灌注组（IR），每组8 只。采用夹闭双侧肾蒂25 min，再灌注48 h 复制大鼠急性肾损伤模型。取大鼠肾脏HE 染色观察病理学结果，血标本测定血尿素氮（BUN）和血肌酐（Scr）水平，比色法测定超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量，末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记法检测细胞凋亡指数，Western blot 检测TXNIP 和炎性体3 蛋白（NLRP3）的表达，酶联免疫吸附测定法检测白细胞介素1β（IL-1β）。结果 与S 组比较，IR 组肾小管肿胀，间质水肿，刷状缘丢失，空泡变性坏死；与S 组比较，IR 组BUN，Scr，MDA，TXNIP，NLRP3 及IL-1β 表达增高（P <0.05），凋亡指数增高（P <0.05），SOD 活性降低（P <0.05）。结论 TXNIP 可能通过激活NLRP3/IL-1β 炎症通路参与大鼠急性肾损伤。

Abstract:

Objective To investigate the role of thioredoxin-interacting protein (TXNIP) in rat model of acute renal injury. Methods Renal ischemia reperfusion (IR) model was established by bilateral occlusion of renal pedicle for 25 min followed by 48 hours' reperfusion. Totally 16 male Sprague-Dawley rats were randomly divided into 2 groups (n = 8): Sham (S) group and IR group. Kidneys were harvested at the 48th h post reperfusion. HE staining was performed for morphological abnormalities. Blood urea nitrogen (BUN) and Serum creatinine (Scr) were measured by fully automatic biochemical analyzer. Concentration of Superoxide Dismutase (SOD) and Malondialdehyde(MDA) were determined by colorimetry. Cellular apoptosis rate was identified by TdT-mediated dUTP Nick-End Labeling. Expression levels of TXNIP, Nod-like receptor protein 3 (NLRP3) and IL-1β were determined by Western blot or Enzyme-linked immunosorbent assay (ELISA). Results Morphological abnormalities such as tissue swelling,interstitial edema, loss of brush border and the necrotic cavitation in renal tubules of IR group were observed. Levels of BUN, Scr, MDA, ,TXNIP, NLRP3, IL-1β as well as cellular apoptosis rate in IR group increased significantly when compared with Sham group (P < 0.05), while concentration of SOD decreased in IR group in comparison with Sham group (P < 0.05). Conclusion TXNIP is involved in renal ischemia reperfusion injury by activing NLRP3/IL-1β inflammatory pathway.