丹参酮IIA(TanIIA)对脑缺血再灌注损伤(CIRI)具有神经保护作用,但由于水溶性差和强大的首过消除性能,其临床应用受到限制。在这项研究中,我们开发了载有TanIIA(TanIIAME)的微乳液来突破这些限制,并探讨了TanIIAME对CIRI的神经保护作用以及这种神经保护的表观遗传调控机制。在体内,用TanIIAME和TanIIA溶液或丙戊酸钠治疗大脑中动脉闭塞(MCAO)模型。通过ELISA测定法测定TanIIAME对HDAC活性的影响。此外,我们使用原代海马神经元建立氧糖剥夺和复氧(OGD/R)模型。进行乳酸脱氢酶(LDH)测定和末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)测定以研究TanIIAME的神经保护功效。随后,H3K18ac的表达,H4K8ac,在用TanIIAME处理的MCAO或OGD/R模型中研究了NMDAR1,caspase-3和MAP-2,TanIIA溶液或丙戊酸钠。体内实验结果表明,TanIIAME显著降低了神经评分,梗死体积,和HDAC活性与TanIIA溶液和MCAO组相比,伴随着H3K18ac的上调,H4K8ac,MAP-2的表达和NMDAR1和caspase-3的表达下调。此外,在OGD/R模型中,结果表明,随着H3K18ac的增加,TanIIAME治疗具有更好的神经保护作用,H4K8ac,和MAP-2表达和减少NMDAR1和caspase-3表达,与除丙戊酸钠外的其他治疗方法相比。总的来说,TanIIAME治疗通过可能涉及抑制NMDAR1和caspase-3表达以及通过调节组蛋白H3K18和H4K8乙酰化增强MAP-2表达的机制,在保护抗CIRI方面表现出优异的功效。因此,TanIIAME可能用于开发一种有前途的治疗缺血性中风的药物。
Tanshinone IIA (TanIIA) has neuroprotective effects against cerebral ischemia reperfusion injury (CIRI), but its clinical application is limited due to poor water solubility and robust first pass elimination property. In this study, we developed microemulsion loaded with TanIIA (TanIIA ME) to break through these limitations, and explored the neuroprotective effect of TanIIA ME against CIRI and the epigenetic regulation mechanism of this neuroprotection. In vivo, middle cerebral artery occlusion (MCAO) models were treated with TanIIA ME and TanIIA solution or sodium valproate as a control. The effect of TanIIA ME on HDAC activity was determined by ELISA assay. In addition, we used primary hippocampal neurons to establish oxygen-glucose deprivation and reoxygenation (OGD/R) models. Lactate dehydrogenase (LDH) assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were performed to investigate the neuroprotective efficacy of TanIIA ME. Subsequently, the expression of
H3K18ac, H4K8ac, NMDAR1, caspase-3, and MAP-2 were investigated in MCAO or OGD/R models treated with TanIIA ME, TanIIA solution or sodium valproate. In vivo experimental results indicated that TanIIA ME significantly reduced neurological scores, infarction volume, and HDAC activity compared with TanIIA solution and MCAO group, accompanied by upregulation of
H3K18ac, H4K8ac, and MAP-2 expression and downregulation of NMDAR1 and caspase-3 expression. Additionally, in OGD/R models, the results demonstrated that TanIIA ME treatment had a better neuroprotective effect along with increased
H3K18ac, H4K8ac, and MAP-2 expression and decreased NMDAR1 and caspase-3 expression, compared with the other treatments except sodium valproate. Overall, TanIIA ME treatment exhibited superior efficacy in protecting against CIRI through mechanisms that might involve the inhibition of NMDAR1 and caspase-3 expression and the enhancement of MAP-2 expression by regulating histone H3K18 and H4K8 acetylation. Thus, TanIIA ME could be potentially used to develop a promising drug for the treatment of ischemic stroke.