甲基莲心碱对脑缺血再灌注脑损伤的调节作用

Regulatory Effect of of Neferine on Brain Tissue Injury of Cerebral Ischemia-Reperfusion

目的探讨甲基莲心碱(Neferine,Nef)对缺血再灌注脑损伤大鼠脑保护作用及其机制.方法线栓法建立大鼠脑缺血再灌注损伤(CI/R)模型.造模后对大鼠神经功能进行评分;HE检测病理损伤;检测血清MDA、SOD和LDH含量;TUNEL检测脑细胞凋亡;RT-PCR检测Caspase-3、-9 mRNA表达;免疫组化检测脑组织ICAM-1表达;ELISA检测IL-6、IL-18和IL-1β的含量;蛋白印迹检测TLR4、NF-κB P65(核)和MIP-2表达.结果甲基莲心碱能有效改善脑缺血再灌注损伤大鼠的神经功能(P<0.01),减少造模诱导的脑组织病理损伤;减少MDA和LDH含量(P<0.01),增加SOD活性(P<0.01);减少脑组织细胞凋亡率(P<0.01),下调凋亡相关蛋白Caspase-3、-9 mRNA表达水平(P<0.01);降低促炎因子(ICAM-1)、炎性因子(IL-6,IL-18,IL-1β)和炎性蛋白(TLR4,NF-κB P65,MIP-2)在脑组织中的表达水平(P<0.01).结论甲基莲心碱能对脑缺血再灌注损伤大鼠脑组织起保护作用,其机制与减少凋亡及抑制炎性反应有关.

Objective To investigate the protective effect of Neferine on brain tissue of rats with cerebral ischemia-reperfusion injury and the mechanism.Methods Ischemia-reperfusion injury(CI/R)of rats was induced by suture method.We evaluated the neurological function and patholog-ical injury of cerebral tissues in rats after model establishment,measured the serum levels of MDA,SOD and LDH,detected apoptosis and mRNA levels of Caspase-3,-9 by using TUNEL and RT-PCR,and determined the expression of ICAM-1 and IL-6,IL-18,IL-1βby immunohistochemical assay and ELISA.Western blotting was performed to detect the expression of TLR4,NF-kappa B P65(nucleus)and MIP-2.Results Neferine could effectively improve the neurological function in rats with cerebral ischemia-reperfusion injury(P<0.01),reduce the pathological damage of brain tissue induced by CI/R,decrease the content of MDA and LDH(P<0.01),increase the activity of SOD(P<0.01),reduce the apoptotic rate of brain tissue(P<0.01),down-regulate the mR-NA expression of apoptotic-related protein Caspase-3 and-9(P<0.01),lower the levels of proin-flammatory factors(ICAM-1),inflammatory factors(IL-6,IL-18,IL-1 beta)and inflammatory proteins(TLR4,NF-kappa P65,MIP-2)in brain tissues(P<0.01).Conclusion Neferine can effectively protect brain tissue of CI/R rats from cerebral ischemia-reperfusion injury,and the mechanism is related to reduced apoptosis and inhibited inflammation.