鸢尾素对新生大鼠高胆红素血症抗炎作用的研究

The anti-inflammatory effect of irisin on hyperbilirubinemia in neonatal rats

目的:探讨鸢尾素干预高胆红素血症新生大鼠脑组织炎症反应的影响。方法:将66只新生SD(Sprague-Dawley)大鼠,采用抽签法随机分为正常对照组(A组)和模型组(M组),于7日龄、10日龄采用腹腔注射胆红素溶液的方法建立高胆红素血症模型。末次注射后,从A组、M组中随机选取6只大鼠,用于验证模型建立成功,然后将M组随机分为B组与C组,C组大鼠侧脑室注射80μg/kg剂量鸢尾素溶液,余各组注射等量磷酸盐缓冲液。侧脑室注射后,根据不同处死点,将A、B、C各组进一步分为3个亚组,在各时间点进行神经行为学检测,苏木精-伊红染色(hematoxylin-eosin staining,HE)检测皮质或海马神经细胞形态,酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测肿瘤坏死因子(tumor necrosis factor-alpha,TNF-α)、白细胞介素1β(interleukin-1 beta,IL-1β)、白细胞介素-6(interleukin,IL-6)的浓度。结果:平面翻正反射:12h时,3组大鼠所需时间无显著性差异(P>0.05);24h时,B、C组大鼠长于A组大鼠(P<0.05);48h时,B组大鼠长于A、C组大鼠(P<0.05),A、C2组大鼠无显著性差异(P>0.05)。各时间点3组大鼠方向趋向性试验均无显著性差异(P>0.05)。HE:A组大鼠皮质神经元结构清晰完整,但海马神经元数量出现减少;B、C组大鼠皮质、海马等区域神经元数目减少,结构紊乱,可见不同程度病理损害,C组优于B组。Elisa:B组脑组织各时间点TNF-α、IL-1β、IL-6的浓度均高于A组(P<0.05);C组脑组织各时间点TNF-α、IL1β、IL-6的浓度均低于B组,差异具有显著性意义(P<0.05);C组大鼠脑组织TNF-α浓度在12h时高于A组大鼠(P<0.05),在24h、48h低于A组大鼠(P<0.05);C组大鼠脑组织IL-6的浓度在12h、24h均低于A组大鼠(P<0.05),48h时,C组大鼠脑组织IL-6的浓度与A组大鼠无显著性差异(P>0.05);C组大鼠脑组织IL-β的浓度在3个时间点均与A组大鼠无显著性差异(P>0.05)。结论:鸢尾素可有效减少高胆红素血症新生大鼠脑组织TNF-α、IL-1β、IL-6的释放,发挥抗炎作用。

Objective:To investigate the effect of irisin on inflammatory response in brain tissue of neonatal rats with hyperbilirubinemia.Method:Sixty-six neonatal SD rats were randomly divided into the normal control group(group A) and the model group(group M).Hyperbilirubinemia models were established by intraperitoneal injection of bilirubin solution at the age of 7 days and 10 days.After the last injection,6 rats were randomly selected from group A and M to verify the success of the model establishment,and then group M was randomly divided into group B and group C.Rats in group C were injected with 80 μg/kg irisin solution in lateral ventricles,and the other groups were injected with the same amount of PBS.After the injection into lateral ventricle,groups A,B and C were further divided into three subgroups according to different points of death.Neurobehavioral tests were performed at each time point.Morphology of cortical or hippocampal nerve cells was detected by HE staining,and the concentrations of TNF-α,IL-1β and IL-6 were detected by ELISA.Result:At 12h,there was no significant difference in the time of plane righting reflex among the three groups(P>0.05);At 24h,the rats in group B and C were longer than those in group A(P>0.05);At 48h,group B was longer than group A and C(P<0.05),and there was no significant difference between group A and C(P>0.05).There was no significant difference in tropism test among the three groups at each time point(P>0.05).Under the HE staining,the cortex neuron structure in group A were clear and complete but the number of hippocampal neurons was reduce;in group B and C,the number of neurons in cortex and hippocampus was decreased,the structure was disordered,and different degrees of pathological damage were observed.Group C was better than group B.Under the ELISA,the concentrations of TNF-α,IL-1β and IL-6 in group B were higher than those in group A at each time point(P>0.05);the concentrations of TNF-α,IL1βand IL-6 in group C were significantly lower than those in group B at each time point(P>0.05);the concentration of TNF-α in the brain tissue of group C was higher than that of group A at 12h(P>0.05),and at24h and 48h were lower than those in group A(P>0.05);the concentration of IL-6 in the brain tissue of group C was lower than that of group A at 12h and 24h(P>0.05);at 48h,there was no significant difference in the concentration of IL-6 between group C and group A(P>0.05);the concentration of IL-1β in the brain tissue of group C was not significantly different from that of group A at the three time points(P>0.05).Conclusion:Irisin can effectively reduce the release of TNF-α,IL-1β and IL-6 in the brain tissue of neonatal rats with hyperbilirubinemia,and play an anti-inflammatory role.