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梁意萍, 付文, 张振海, 吕慧侠. 三七总皂苷玉米脂蛋白纳米粒对大鼠脑缺血再灌注损伤的药效学研究[J]. 中国药科大学学报, 2021, 52(3): 318-324. DOI: 10.11665/j.issn.1000-5048.20210308
引用本文: 梁意萍, 付文, 张振海, 吕慧侠. 三七总皂苷玉米脂蛋白纳米粒对大鼠脑缺血再灌注损伤的药效学研究[J]. 中国药科大学学报, 2021, 52(3): 318-324. DOI: 10.11665/j.issn.1000-5048.20210308
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LIANG Yiping, FU Wen, ZHANG Zhenhai, LYU Huixia. Pharmacodynamics study of Panax notoginseng saponins zein lipoprotein nanoparticles for cerebral ischemia reperfusion injury in rats[J]. Journal of China Pharmaceutical University, 2021, 52(3): 318-324. DOI: 10.11665/j.issn.1000-5048.20210308
Citation: LIANG Yiping, FU Wen, ZHANG Zhenhai, LYU Huixia. Pharmacodynamics study of Panax notoginseng saponins zein lipoprotein nanoparticles for cerebral ischemia reperfusion injury in rats[J]. Journal of China Pharmaceutical University, 2021, 52(3): 318-324. DOI: 10.11665/j.issn.1000-5048.20210308
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三七总皂苷玉米脂蛋白纳米粒对大鼠脑缺血再灌注损伤的药效学研究

Pharmacodynamics study of Panax notoginseng saponins zein lipoprotein nanoparticles for cerebral ischemia reperfusion injury in rats

    摘要
  • 摘要: 对三七总皂苷（Panax notoginseng saponins，PNS）与卵磷脂、β-谷甾醇、玉米醇溶蛋白共组装制备的三七总皂苷玉米脂蛋白纳米粒（PNS-lipid-zein nanoparticles，PLZ-NPs）进行了体外细胞实验，以及大鼠脑缺血再灌注损伤的药效学研究。采用反溶剂沉淀法制备了PLZ-NPs，运用激光粒度仪和透射电镜显微镜对纳米粒进行表征，MTT法评价了纳米粒的细胞毒性，激光共聚焦和流式细胞术分析了Caco-2细胞对纳米粒的摄取情况。大脑中动脉闭塞（MCAO）法构建了大鼠脑缺血再灌注损伤模型，连续3 d灌胃给药后，取脑组织进行2，3，5-氯化三苯基四氮唑（TTC）染色，并检测脂质过氧化物丙二醛（MDA）、炎症因子IL-1β、TNF-α、凋亡相关蛋白Bax、Bcl-2的水平。结果显示，PLZ-NPs平均粒径为（116.4 ± 0.81）nm，PDI为0.048，Zeta电位为-（31.5 ± 0.31）mV。MTT实验表明，玉米脂蛋白载体的生物安全性良好，且能显著提高Caco-2细胞对纳米粒的摄取（纳米粒组在4 h时细胞摄取量提高至游离组的1.76倍）。TTC染色中，PNS和PLZ-NPs组的梗死面积比模型组均有减少，另外MDA、IL-1β、TNF-α、Bax的含量明显下降，Bcl-2含量增加，而且PLZ-NPs组对脑缺血再灌注损伤的保护作用显著优于PNS组（P<0.05）。实验结果表明，本实验制备的玉米脂蛋白纳米粒具有良好的稳定性、药物负载能力和生物安全性，能显著提高肠上皮细胞对药物的摄取，有效改善脑缺血再灌注对大鼠造成的损伤。

     

    Abstract: PLZ-NPs (PNS-lipid-zein nanoparticles) prepared by co-assembly of Panax notoginseng saponins, lecithin, β-sitosterol and zein were applied for in vitro cell experiment and oral gavage to study the protective effect of cerebral ischemia-reperfusion rats.PLZ-NPs were characterized by Malvin-particle size analyzer and transmission electron microscope (TEM), respectively. The toxicity of PLZ-NPs and free carrier were evaluated by MTT, and the uptake of nanoparticles in Caco-2 cells was analyzed by laser confocal and flow cytometry. The cerebral ischemia reperfusion rat model was established by MCAO method and then be given samples by gavage for 3 days. The brain tissues were taken to stain by 2, 3, 5-triphenyltetrazole chloride (TTC) and the biochemical indicators of MDA, inflammatory cytokines IL-1β and TNF-α, apoptosis-related proteins Bax and Bcl-2 from the harvested brain tissues were detected to evaluate the protective effect of PNS in PLZ-NPs on cerebral ischemia reperfusion. The particle size, PDI, and zeta potential of formed PLZ-NPs were (116.4 ± 0.81) nm, 0.048 and -(31.5 ± 0.31) mV, respectively. The results of MTT showed that the zein lipoprotein carrier was non-toxic to Caco-2 cells. The results of laser confocal and flow cytometry showed that FITC uptake of nanoparticles could be significantly improved in Caco-2 cells.The uptake from the nanoparticles at 4h was 1.76 times of that of the free FITC group.Compared with the model group, the TTC staining images of free drug PNS group and PLZ-NPs group showed certain reduction in the white infarct area.The contents of MDA, IL-1β, TNF-α and Bax were significantly decreased, while the content of Bcl-2 was significantly increased. Furthermore, all parameters of PLZ-NPs group showed better results than those of PNS group, and there was a significant difference (P < 0.05). All results indicated that the prepared PLZ-NPs had good stability and biological safety, and could significantly increase the uptake in intestinal epithelial cells, and effectively protect against the damage caused by cerebral ischemia reperfusion in rats.

     

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