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目的探讨巨噬细胞移动抑制因子(MIF)在缺氧缺血性脑损伤(HIBD)新生大鼠中的作用及可能机制。方法健康7日龄新生SD大鼠共120只。随机分为3组,分别为假手术组、HIBD组、抗MIF中和抗体干预组(anti-MIF组,缺氧缺血后即予抗MIF中和抗体5 mg.kg-1腹腔注射),每组40只。于HIBD模型制成后6 h、12 h、24 h,3 d及7 d处死,应用免疫组织化学法(SP法)观察各组脑缺血侧皮质区MIF及核因子-κB(NF-κB)蛋白的表达,双抗夹心酶联免疫吸附法(ELISA)测定各组缺血侧脑组织匀浆TNF-α水平。结果 HIBD组新生大鼠脑组织皮质区MIF、TNF-α表达均在缺氧缺血6 h明显增加,24 h达高峰(Pa<0.01),7 d时恢复正常,与假手术组比较差异无统计学意义(P>0.05);HIBD组NF-κB表达在缺氧缺血6 h开始增加,24 h达高峰,并维持至3 d(Pa<0.01),7 d降至正常,与假手术组比较差异无统计学意义(P>0.05);anti-MIF组大鼠脑组织MIF、NF-κB、TNF-α的表达与HIBD组比较,在各个实验时间点均有降低(Pa<0.01)。结论 MIF可能通过调控NF-κB的途径影响其脑皮质TNF-α水平,从而介导缺氧缺血后新生大鼠脑组织的炎性损伤。
Objective To investigate the role and possible mechanism of macrophage migration inhibitory factor (MIF) in neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods A total of 120 newborn SD rats aged 7 days were enrolled. The rats were randomly divided into 3 groups: sham operation group, HIBD group, anti-MIF neutralizing antibody intervention group (anti-MIF group, intraperitoneal injection of anti-MIF neutralizing antibody 5 mg.kg-1) 40 per group The HIBD models were sacrificed at 6 h, 12 h, 24 h, 3 d and 7 d after operation. Immunohistochemistry (SP method) was used to observe the expression of MIF and NF-κB ) Protein were detected by enzyme-linked immunosorbent assay (ELISA). TNF-α levels in the ischemic brain homogenate of each group were measured by enzyme-linked immunosorbent assay (ELISA). Results The expression of MIF and TNF-α in cortex of neonatal rats in HIBD group increased significantly at 6 h of hypoxia and ischemia, peaked at 24 h (P <0.01), returned to normal at 7 d, and no significant difference compared with sham operation group (P> 0.05). The expression of NF-κB in HIBD group began to increase at 6 h and reached the peak at 24 h, maintained at 3 d (Pa <0.01), and decreased to normal at 7 d (P> 0.05). Compared with HIBD group, the expression of MIF, NF-κB and TNF-α in anti-MIF group decreased at each experimental time point (P <0.01) . Conclusions MIF may affect the level of TNF-α in cerebral cortex by regulating the pathway of NF-κB, and thus mediate the inflammatory injury of neonatal rat brain after hypoxic-ischemic brain damage.