中性鞘磷脂酶-2(Neutral sphingomyelin-2 N-SMase2)对大鼠脑缺血再灌注损伤中IL-6、IL-1β、TNF-α炎性因子的影响

Effects of neutral sphingomyelinenzyme-2 on IL-6,IL-1β and TNF-α inflammatory factors in rats with cerebral ischemia reperfusion injury

目的研究中性鞘磷脂酶-2(Neutral sphingomyelin-2 N-SMase2)对大鼠脑缺血再灌注损伤中IL-6、IL-1β、TNF-α炎性因子的影响。方法采用线栓法制作SD大鼠大脑中动脉缺血再灌注(ischemia reperfusion I/R)损伤模型,侧脑室注射N-SMase2激活剂(daunorubicin DNR)或抑制剂GW4869,免疫组织化学技术检测蛋白神经酰胺(Ceramide Cera)和N-SMase2的表达,RT-PCR技术检测IL-6、IL-1β、TNF-α等炎性因子的mRNA表达水平。结果与对照组相比,再灌注组(I/R)大鼠脑组织Ceramide和N-SMase2的蛋白表达阳性细胞较对照组增多(P<0.05),IL-6、IL-1β、TNF-α炎性因子的mRNA表达水平升高(P<0.05),侧脑室注射GW4869可显著降低IL-6、IL-1β、TNF-α的mRNA水平(P<0.05)。结论脑缺血再灌注模型中,N-SMase/Cera通路激活,参与调控神经细胞再灌注损伤,而N-SMase2抑制剂GW4869可明显降低IL-6、IL-1β、TNF-α炎性因子的表达。

Objective To study of the effects of neutral sphingomyelin-2 on inflammatory factors of IL-6,IL-1β,TNF-α in rats with cerebral ischemia reperfusion injury. Methods Focal cerebral ischemia reperfusion model rats were subjected to MCAO（ Middle Cerebral Artery Occlusion）. The rats were pretreated by injection of the N-SMase activator daunorubicin（ DNR） or inhibitor GW4869 into their lateral ventricles in the ischemia reperfusion group. The expressions of ceramide and N-SMase2 proteins was were detected by immunohistochemical technique. The mRNA expressions of IL-6,IL-1β,TNF-α in ischemic reperfusion region were investigated by real-time PCR. Results Compared with the sham group,the expressions of Ceramide and N-SMase2 proteins increased in the ischemia reperfusion group（ P〈0. 05）. The mRNA expressions of IL-6,IL-1β,TNF-α increased significantly in the ischemia reperfusion group（ P〈0. 05）. N-SMase inhibitor GW4869 attenuated the mRNA expressions of IL-6,IL-1β,TNF-α（ P〈0. 05）. Conclusion The activation of N-SMase/Ceramide pathway participates in the process of cerebral ischemia reperfusion inflammatory injury. However,N-SMase2 inhibitor GW4869 can significantly reduce the expression of IL-6,IL-1β,TNF-α inflammatory factors.