黑磷纳米片载药体系的制备及其抗缺血性脑损伤的作用

Preparation of drug loading system of black phosphorus nanosheets and its anti-ischemic brain injury

本研究采用普通液相法制备黑磷纳米片(black phosphorus nanosheets,BP),并经聚乙二醇修饰后将白藜芦醇负载于BP上,得BP载药体系。采用荧光蛋白标记法考察BP的脑靶向性,并通过TTC(2,3,5-triphenyltetrazolium chloride)染色法、神经行为学评价、脑水肿考察等实验研究黑磷载药后抗脑缺血/再灌注脑损伤的作用。使用蛋白免疫印迹分析方法探究BP载药体系作用于缺血性脑损伤的分子机制,采用溶血实验、苏木精-伊红(hematoxylin-eosin,H&E)染色实验评估其生物相容性。结果表明,BP载药能力优异,其载药体系结构、粒径均一且释药曲线稳定,具有优异的光热效应。通过对荧光强度的分析对比,发现BP在近红外光辅助照射条件下可增加血脑屏障通透性,使药物更多通过血脑屏障。此外,BP载药体系能明显改善造模后小鼠的神经行为障碍,脑梗死面积及脑水肿程度显著下降。蛋白免疫印迹分析实验证明,此载药体系可通过激活抗氧化信号通路蛋白核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)、血红素氧合酶-1(heme oxygenase-1,HO-1)的表达,从而发挥抗缺血性脑损伤作用。BP载药体系的溶血实验及H&E实验结果表明,其不具有明显毒性,安全性高。综上,本研究制备的BP载药量高、光热性能良好、安全性高,在近红外光条件下,还具有一定的脑靶向能力,可提高药物在脑部的治疗效果。动物福利和实验过程均遵循石河子大学一附院动物伦理委员会的规定。

In this study,black phosphorus nanosheets(BP) were prepared by the ordinary liquid phase method,and resveratrol was loaded on the BP after being modified by polyethylene glycol.The brain targeting of BP was investigated by fluorescent protein labeling,and the effects of black phosphorus on cerebral ischemia/reperfusion injury were studied by 2,3,5-triphenyltetrazolium chloride(TTC) staining,neurobehavioral evaluation,and brain edema.Protein immunoblotting analysis was used to explore the molecular mechanism of the BP drug delivery system on ischemic brain injury.Hemolysis test and hematoxylin-eosin(H&E) staining were used to evaluate its biocompatibility.The results showed that BP had excellent drug loading capacity,uniform drug loading system structure and particle size,stable drug release curve,and excellent photothermal effect.Through the analysis and comparison of fluorescence intensity,it was found that BP can increase the permeability of blood-brain barrier(BBB) under the condition of near-infrared light assisted irradiation,and make drugs more pass through the BBB.In addition,the black phosphorus nano tablet drug delivery system can significantly improve the neurobehavioral disorder of mice after modeling,and the cerebral infarction area and brain edema degree are significantly decreased.Western blot experiments showed that the drug delivery system could play an anti-ischemic brain injury role by activating the expression of antioxidant signaling pathway proteins nuclear factor E2-related factor 2(Nrf2) and heme oxygenase-1(HO-1).The hemolysis test and H&E test results of the BP drug carrier system showed that it had no obvious toxicity and high safety.In conclusion,the BP prepared in this study had high drug loading,good photothermal performance,and high safety.Under the near-infrared condition,they also have certain brain targeting ability,which can improve the therapeutic effect of drugs in the brain.Animal welfare and experimental procedures were following the regulations of the Animal Ethics Committee of the First Affiliated Hospital of Shihezi University.