基于PI3K/Akt通路豨莶草抗脑缺血再灌注损伤作用机制研究

Study the mechanism of Herba Siegesbeckiae for cerebral ischemia-reperfusion based on PI3K/Akt pathway

目的基于PI3K/Akt通路研究豨莶草抗脑缺血再灌注(IR)损伤的作用机制。方法将40只SD大鼠随机分成假手术组、模型组和豨莶草不同剂量(0.45、0.90、1.35 g/kg)组,每组8只,各组给予相应的药物,连续灌胃给药14 d。IR 24 h后,对各组大鼠进行神经功能缺损评分和病理学检查,ELISA法检测缺血脑组织中IL-1β、IL-6、TNF-α和NF-κB含量,RT-PCR和Western-blot技术检测缺血脑组织中PI3K、Akt mRNA和蛋白的表达。结果与假手术组相比,模型组大鼠神经功能缺损评分明显降低(P<0.01),梗死灶周围炎性细胞浸润程度和范围显著增加,缺血脑组织中PI3K、Akt的mRNA和蛋白表达明显升高(均P<0.01),IL-1β、IL-6、TNF-α和NF-κB含量显著升高(均P<0.01);0.90 g/kg和1.35 g/kg豨莶草能显著降低IR模型大鼠神经功能缺损评分(均P<0.01),梗死灶周围炎性细胞浸润程度和范围显著减少,PI3K、Akt的mRNA和蛋白表达明显减少(均P<0.01),IL-1β、IL-6、TNF-α和NF-κB含量明显降低(均P<0.01)。结论豨莶草抗IR损伤可能是通过调节PI3K/Akt信号通路抑制NF-κB、TNF-α、IL-1β和IL-6含量实现的。

Objective To study the mechanism of Herba Siegesbeckiae for cerebral ischemia reperfusion(IR)injury based on PI3K/Akt pathway.Methods A total of 40 SD rats were randomly divided into sham operation group,model group and Herba Siegesbeckiae group(0.45,0.90,1.35 g/kg),8 rats in each group,and different medication intervention were administrated for 14 d according to the groups.After 24 hours of IR,neurofunctional defect scores and pathological examinations were performed,RTPCR method and western-blot were used to detect the mRNA and protein expression of PI3K and Akt,the levels of IL-1β,IL-6,TNF-αand NF-κB were detected by ELISA.Results Compared with the sham group,the neurofunctional defect scores in the model group were significantly increased(all P<0.01),and the degree and range of inflammatory cell infiltration were significantly increased(all P<0.01),and the mRNA and protein expressions of PI3K and Akt were significantly increased(all P<0.01),and the levels of IL-1β,IL-6,TNF-αand NF-κB were significantly increased(all P<0.01);while all of the above results could be reversed by 0.90 g/kg and 1.35 g/kg Herba Siegesbeckiae(all P<0.01).Conclusion Herba Siegesbeckiae could reduce the injury induced by IR through regulating the PI3K/Akt signaling pathway to inhibit the release of IL-1β,IL-6,TNF-αand NF-κB.