山柰酚对缺血性脑损伤大鼠的神经保护作用研究

Neuroprotective effects of kaempferol on ischemic brain injury in rat

目的探讨山柰酚(Kaempferol,Kae)对缺血性脑损伤大鼠的神经保护作用。方法山柰酚连续灌胃7 d后进行大脑中动脉栓塞(Middle cerebral artery occlusion,MCAO),缺血2 h后拔出线栓进行再灌注24 h后对大鼠进行神经功能评分;红四氮唑染色测定大鼠脑梗死面积比例; ELISA试剂盒检测梗死侧海马组织ROS、MDA和SOD的含量; Real time PCR检测TNF-α、IL-1β和IL-6 mRNA表达;免疫印迹检测海马组织NF-κB p65、Bax和Bcl-2蛋白表达。结果山柰酚中、高剂量明显提高MCAO小鼠神经功能受损评分,减少脑梗死面积比例(P <0. 05,P <0. 01); ELISA结果显示,山柰酚各剂量均可显著性增加MCAO小鼠海马SOD含量,降低其ROS和MDA的含量;Real time PCR和免疫印迹结果显示,山柰酚能明显减少MCAO小鼠海马TNF-α、IL-1β和IL-6 mRNA表达,降低NF-κB p65、Bax的表达及增加Bcl-2蛋白表达。结论山柰酚对缺血性脑损伤的神经保护作用与其抗氧化、抗炎及抗凋亡作用密切相关。

Objective To investigate the neuroprotective effects of kaempferol on ischemic brain injury in rat.Method Rats were intragastricly treated with kaempferol or vehicle for consecutive 7 days prior to ischemia.Middle cerebral artery were occlude for 2 h and then reperfused for 24 h followed by neurological deficit score.TTC staining was used to detect the infarct volume of brain.The ROS,MDA and SOD contents were determined by ELISA analysis.Real time PCR analysis was used to examine the mRNA expression of TNF-α,IL-1βand IL-6 in hippocampus of rats.The protein expression of NF-κB p65,Bax and Bcl-2 were examined by Western blotting.Result Kaempferol significantly increased the neurological deficit scores and decreased cerebral infarct volume in MCAO rats at middle and high doses(P<0.05,P<0.01).ELISA analysis indicated kampferol significantly increased SOD content,decreased ROS and MDA contents in hippocampus of MCAO rats.Real time PCR suggested that kampferol decreased TNF-α,IL-1βand IL-6 mRNA expression in hippocampus of MCAO rats.Western blotting analysis showed that kaempferol significantly decreased NF-κB p65 and Bax protein expression,increased Bcl-2 protein expression in hippocampus of MCAO rats.Conclusion The neuroprotective effects of kaempferol on ischemic brain injury are highly related to its anti-oxidation,anti-inflammtion and anti-apoptosis.