5-BDBD对帕金森病模型大鼠脑IL-1β和IL-18表达的影响

EFFECT OF 5-BDBD ON THE EXPRESSION OF INTERLEUKIN-1Β AND INTERLEUKIN-18 IN A RAT MODEL OF PARKINSON'S DISEASE

目的探讨5-BDBD对6-OHDA诱导的帕金森病(PD)模型大鼠脑中白细胞介素1β(IL-1β)和白细胞介素18(IL-18)表达的影响。方法将Wistar大鼠随机分为空白对照组、6-OHDA组、5-BDBD组和5-BDBD+6-OHDA组,每组20只大鼠。5-BDBD经脑立体定位仪定向注射于大鼠左侧侧脑室,预处理1周后,根据分组情况经脑立体定位仪向大鼠左侧黑质定位注射6-OHDA或等量抗坏血酸。造模4周后断头取脑,采用免疫荧光染色法检测各组大鼠脑黑质中多巴胺能神经元,采用Western-blot法检测各组大鼠黑质P2X4受体(P2X4R)、IL-1β和IL-18蛋白的相对表达量。结果与空白对照组及5-BDBD组比较,6-OHDA组、5-BDBD+6-OHDA组大鼠黑质中TH阳性多巴胺能神经元数显著减少(F=162.7,P<0.01),P2X4R、IL-1β和IL-18蛋白的相对表达量显著升高(F=133.4~552.6,P<0.01)。与6-OHDA组比较,5-BDBD+6-OHDA组大鼠黑质中TH阳性多巴胺能神经元数显著增加(F=162.7,P<0.01),P2X4R、IL-1β和IL-18蛋白的相对表达量显著降低(F=133.4~552.6,P<0.01)。结论抑制P2X4R可以抑制炎症反应,减少PD大鼠黑质中TH阳性多巴胺能神经元的变性死亡,并可以抑制6-OHDA诱导的IL-1β和IL-18的表达上调,具有潜在的神经保护作用。

Objective To investigate the effect of 5-BDBD on the expression of interleukin-1β（IL-1β）and interleukin-18（IL-18）in a rat model of Parkinson＇s disease（PD）induced by 6-OHDA. Methods Wistar rats were randomly divided into blank control group,6-OHDA group,5-BDBD group,and 5-BDBD＋6-OHDA group,with 20 rats in each group.5-BDBD was injected into the left lateral ventricle of the rats under a stereotaxic apparatus.After one week of pretreatment,6-OHDA or an equal volume of ascorbic acid was injected into the left substantia nigra under a stereotaxic apparatus.The rats were decapitated at four weeks after the model was established and the brain tissue was collected.Immunofluorescent staining was used to measure dopaminergic neurons in the substantia nigra,and Western-blot was used to measure the relative expression of P2 X4 receptor（P2 X4 R）,IL-1β,and IL-18 in the substantia nigra. Results Compared with the blank control group and the 5-BDBD group,the 6-OHDA group and the 5-BDBD＋6-OHDA group had a significant reduction in the number of tyrosine hydroxylase（TH）-positive dopaminergic neurons in the substantia nigra（F=162.7,P〈0.01）and significant increases in the relative expression of P2 X4 R,IL-1β,and IL-18（F=133.4-552.6,P〈0.01）.Compared with the 6-OHDA group,the 5-BDBD＋6-OHDA group had a significant increase in the number of TH-positive dopaminergic neurons in the substantia nigra（F=162.7,P〈0.01）and significant reductions in the relative expression of P2 X4 R,IL-1β,and IL-18（F=133.4-552.6,P〈0.01）. Conclusion The inhibition of P2 X4 Rcan inhibit inflammatory response and reduce the degeneration and death of TH-positive dopaminergic neurons in PD rats.It can also inhibit the increase in the expression of IL-1βand IL-18 induced by 6-OHDA and thus exerts a potential neuroprotective effect.