摘要
背景:雷公藤内酯酮作为中药雷公藤的生物活性成分对神经元具有一定的保护作用。目的:探讨雷公藤内酯酮对脑缺血再灌注损伤的影响。方法:(1)细胞实验:将海马神经元(HT22细胞)随机分为对照组、糖氧剥夺/复氧组、糖氧剥夺/复氧+雷公藤内酯酮组、糖氧剥夺/复氧+雷公藤内酯酮+敲低TP53诱导的糖酵解和凋亡因子(si-TIGAR)组、糖氧剥夺/复氧+si-TIGAR组、糖氧剥夺/复氧+雷公藤内酯酮+雷帕霉素组。通过CCK-8法检测HT22细胞活力,Western blotting检测TIGAR、谷胱甘肽过氧化酶4、溶质载体家族7成员11、鞘氨醇激酶1和哺乳动物雷帕霉素靶蛋白的蛋白表达水平,生化试剂盒检测谷胱甘肽、丙二醛含量和亚铁离子的水平。(2)动物实验:将大鼠随机分为假手术组、模型组、雷公藤内酯酮组,后两组制备大脑动脉闭塞/再灌注大鼠模型,雷公藤内酯酮组于建模后采用50 mg/kg雷公藤内酯酮灌胃7 d。给药结束后24 h,采用LONGA法评估大鼠神经功能缺损情况,TTC法观察脑组织梗死情况,TUNEL染色检测凋亡水平,Western blotting检测凋亡相关蛋白表达水平。结果与结论:(1)在细胞水平,与对照组比较,雷公藤内酯酮可恢复糖氧剥夺/复氧处理HT22细胞的活力,抑制细胞凋亡,上调HT22细胞中TIGAR、谷胱甘肽过氧化酶4、溶质载体家族7成员11的表达水平,激活SPHK1/mTOR通路;此外,雷公藤内酯酮还可升高谷胱甘肽含量,降低丙二醛含量和亚铁离子水平;而雷帕霉素处理抵消了雷公藤内酯酮对HT22细胞的保护作用;(2)在动物水平,雷公藤内酯酮明显降低了模型组大鼠脑组织的神经功能缺损、梗死体积和细胞凋亡,并抑制模型组大鼠神经元铁死亡;(3)提示雷公藤内酯酮通过上调TIGAR表达水平,激活SPHK1/mTOR信号来抑制铁死亡,从而减轻脑缺血再灌注损伤。结果提示,雷公藤内酯酮可能是脑缺血再灌注损伤治疗的候选药物。
BACKGROUND:Triptolide,a bioactive component of the traditional Chinese medicine Tripterygium wilfordii,has a certain protective effect on neurons.OBJECTIVE:To investigate the effect of triptolide on cerebral ischemia/reperfusion injury.METHODS:(1)Cell experiment:Hippocampal neurons(HT22 cells)were randomly divided into control group,glucose oxygen deprivation/reoxygenation(OGD/R)group,OGD/R+triptolide group,OGD/R+triptolide+si-TIGAR group,OGD/R+si-TIGAR group,and OGD/R+triptolide+rapamycin group.HT22 cell viability was detected by cell counting kit 8.Tp53-induced glycolysis and apoptosis factors,glutathione peroxidase 4,7 members of the solsolic vector family 11,sphingosine kinase 1(SPHK1)and(mTOR)were detected by western blot assay.Glutathione,malondialdehyde and iron level were detected using the biochemical kit.(2)Animal experiment:Rats were randomly divided into sham surgery group,model group,and triptolide group.Cerebral artery occlusion/reperfusion rat models were prepared in the latter two groups.Rats in the triptolide group were orally administered 50 mg/kg triptolide for 7 days.Twenty-four hours after administration,LONGA method was used to evaluate the neurological impairment of rats,TTC method was used to observe the conditions of cerebral infarction,TUNEL staining was used to detect cell apoptosis,and western blot was performed to detect the expression level of related proteins.RESULTS AND CONCLUSION:(1)At the cellular level,triptolide promoted cell viability and inhibited apoptosis in HT22 cells treated with OGD/R.Triptolide also increased the expression levels of Tp53-induced glycolysis and apoptosis factors,glutathione peroxidase 4,and 7 members of the solsolic vector family 11,activated the SPHK1/mTOR pathway,increased glutathione content,inhibited malondialdehyde content and iron levels.Rapamycin treatment counteracted the protective effect of triptolide on HT22 cells.(2)At the animal level,triptolide significantly reduced neurological deficits,infarct volume,and cell apoptosis,and inhibited neuronal ferroptosis in brain tissue of rats.To conclude,triptolide can inhibit ferroptosis by upregulating the expression level of Tp53-induced glycolysis and apoptosis factors and activating the SPHK1/mTOR signaling,and thereby reduced cerebral ischemia/reperfusion injury.These findings suggest that triptolide may be a candidate drug for the treatment of cerebral ischemia/reperfusion injury.
作者
邹荣基
喻芳芳
王茂林
贾卓鹏
Zou Rongji;Yu Fangfang;Wang Maolin;Jia Zhuopeng(Department of Neurosurgery,The First Affiliated Hospital of Xi’an Medical University,Xi’an 710077,Shaanxi Province,China;Department of Neurology,The First Affiliated Hospital of Xi’an Medical University,Xi’an 710077,Shaanxi Province,China)
出处
《中国组织工程研究》
北大核心
2026年第4期873-881,共9页
Chinese Journal of Tissue Engineering Research
基金
西安市未央区科工局科研资金资助项目(202023),项目参与人:邹荣基。
