三七皂苷R_1-低相对分子质量三七多糖纳米粒的制备及其对SH-SY5Y细胞损伤的保护作用

目的利用水溶性低相对分子质量三七多糖(low molecular weight Panax notoginseng polysaccharide,PNP-L)作为新型载体制备纳米粒,实现对三七皂苷R1的增溶并提高其治疗效果。方法利用SephadexG-100凝胶对三七多糖(Panax notoginseng polysaccharide,PNP)分级纯化,得到相对均质的低相对分子质量三七多糖(PNP-L)。以PNP、PNP-L作为三七皂苷R1的载体,采用乳化溶剂挥发法制备三七皂苷R1-三七多糖(R1-PNP)纳米粒和三七皂苷R1-低相对分子质量三七多糖(R1-PNP-L)纳米粒。测定其载药量及包封率,探索低相对分子质量多糖作为载体的可行性。采用马尔文粒度分析仪对纳米粒粒径、粒度分布及Zeta电位进行测定,差示扫描量热仪(DSC)进行晶型分析,透射电子显微镜(TEM)测定其形态。体外培养人神经母细胞瘤SH-SY5Y细胞,构建缺氧/复氧模型,比较三七皂苷R1、R1-PNP-L物理混合制剂、R1-PNP-L纳米粒对神经细胞损伤的保护作用。结果 R1-PNP-L纳米粒平均粒径(139.17±7.22)nm,多分散指数(PDI)为0.41±0.19,Zeta电位(-19.14±3.14)mV,载药量28.62%,包封率85.85%,将三七皂苷R1溶解度提高了32.30倍,并起到缓释作用。R1-PNP-L纳米粒对SH-SY5Y细胞无明显细胞毒性,对缺氧/复氧模型诱导的SH-SY5Y细胞损伤的保护效果R1-PNP-L纳米粒R1-PNP-L物理混合制剂三七皂苷R1。结论 PNP-L可作为三七皂苷R1的一种良好载体,形成的纳米粒可有效提高三七皂苷R1溶解度并对缺氧/复氧模型诱导的SH-SY5Y细胞损伤具有保护作用。

Preparation of notoginsenoside R1-low molecular weight Panax notoginseng polysaccharide nanoparticles and its protective effect on SH-SY5Y cell injury

Objective To increase the solubility of notoginsenoside R₁ and improve the therapeutic effect, hydrophilic low molecular weight Panax notoginseng polysaccharide (PNP-L) was used as a new carrier to construct the nanoparticles. Methods Sephadex G-100 gel was used to fractionate and purify P. notoginseng polysaccharide (PNP) to obtain uniform PNP-L with low molecular weight. Using PNP and PNP-L as the carrier of notoginsenoside R₁, the notoginsenoside R₁-notoginseng polysaccharide nanoparticles (R₁-PNP nanoparticles) and notoginsenoside R₁-notoginseng polysaccharide nanoparticles with low molecular weight (R₁-PNP-L nanoparticles) were prepared by solvent evaporation method. Determining the drug loading and encapsulation efficiency of nanoparticles, and exploring the feasibility of polysaccharides with low molecular weight as carriers. Malvern particle size analyzer was used to measure the particle size, poly dispersity index (PDI) and Zeta potential. Differential scanning calorimeter (DSC) was used to analyze the crystalline form of the nanoparticles. Transmission electron microscopy was used to observe the surface morphology. SH-SY5Y nerve cells were cultured in vitro and a hypoxia/reoxygenation model was constructed to compare the protection effect of P. notosaponin R₁, R₁-PNP-L physical mixture and R₁-PNP-L nanoparticles. Results The particle size, PDI, Zeta potential, drug loading and encapsulation efficiency of R₁-PNP-L nanoparticles were (139.17±7.22) nm, 0.41±0.19, (-19.14±3.14) mV, 28.62% and 85.85%, respectively. R₁-PNP-L nanoparticles increased the solubility of notoginsenoside R₁by 32.30 times and had slow-release effect. R₁-PNP-L nanoparticles had no cytotoxicity to SH-SY5Y cells. Protection effect on SH-SY5Y cell injury induced by hypoxia/reoxygenation model:R₁-PNP-L nanoparticles > R₁-PNP-L physical mixture > notoginsenoside R₁. Conclusion PNP-L is a good carrier to deliver notoginsenoside R₁, and the prepared nanoparticles can improve the solubility of notoginsenoside R₁ effectively and increase the protection effect of SH-SY5Y cells after hypoxia and reoxygenation injury.