不同提取工艺桔梗多糖的理化性质分析及其对奥沙利铂体内分布的影响

Platycodon grandiflorum Polysaccharides with Dellifferent Extraction Processes and Their Effects on Oxaliplatin Distribution in Vivo

目的:探究不同提取工艺桔梗多糖(PGP)的理化性质,不同提取工艺所得的PGP对奥沙利铂(OXA)体内分布的影响。方法:本研究采用了热水提取(HW)、醋酸辅助提取(CA)、氨水辅助提取(KA)、超声辅助提取(UA)制备桔梗多糖,通过红外光谱、紫外光谱、凝胶色谱、高效液相色谱等方法测定多糖的理化性质;建立OXA电感耦合等离子体-质谱(ICP-MS)体内分析方法,考察OXA单用组、OXA联用PGP-HW组、OXA联用PGP-CA组、OXA联用PGP-KA组、OXA联用PGP-UA组SD大鼠体内奥沙利铂分布的变化;使用Western Blot方法来测定肺组织中Oct-2和Pg-P蛋白的表达水平,考察PGP改变OXA体内分布的机制。结果:四种桔梗多糖在理化性质方面呈现出明显的区别。其中,PGP-KA的得率最高;四种桔梗多糖红外图谱均具备典型的多糖特征吸收峰;在分子量组成上,它们主要由7 kDa和3790 kDa两个分子量构成,但各自的分子量分布比例有所不同。此外,这四种桔梗多糖都含有甘露糖(Man)、鼠李糖(Rha)、半乳糖醛酸(GalA)、葡萄糖(Glu)、半乳糖(Gal)以及阿拉伯糖(Ara),但比例存在差异;四种多糖粒径主要集中在300 nm左右,且具有良好的溶液稳定性;与OXA单用组比,OXA联用PGP-CA组、OXA联用PGP-KA组分别增加肺组织OXA含量为36.93%和35.12%,且OXA联用PGP-KA组显著(P<0.05)增加Oct-2蛋白表达水平,而P-g-P蛋白表达水平无显著差异。结论:不同的提取工艺获得的PGP的理化性质不同,其中OXA联用PGP-CA组和OXA联用PGPKA组能显著增加OXA在肺组织分布水平,其机制可能与增加肺组织中Oct-2的表达量有关。

Objective:To investigate the physicochemical properties of PGP extracted by different processes and the effects of these PGPs on the in vivo distribution of Oxaliplatin(OXA).Methods:In this study,hot water extraction(HW),acetic acid-assisted extraction(CA),ammonia-assisted extraction(KA),and ultrasound-assisted extraction(UA)were used to prepare PGP.The physicochemical properties of the polysaccharides were determined by methods such as infrared spectroscopy,ultraviolet spectroscopy,gel chromatography,and high-performance liquid chromatography.An inductively coupled plasma mass spectrometry(ICP-MS)method was established to analyze the in vivo distribution of OXA in SD rats treated with OXA alone,OXA+PGP-HW,OXA+PGP-CA,OXA+PGP-KA,and OXA+PGP-UA.The expression levels of Oct-2 and Pg-P proteins in lung tissues were measured by Western blot to investigate the mechanism of PGP in modifying the in vivo distribution of OXA.Results:Significant differences existed in the physicochemical properties of the four types of PGPs.Among these four types,PGP-KA had the highest yield.The infrared spectra of all four PGPs exhibited typical absorption peaks characteristic of polysaccharides.The four PGPs consisted primarily of two molecular weights,7 kDa and 3790 kDa,but the distribution percentages of these molecular weights vary.All four PGPs contained mannose(Man),rhamnose(Rha),galacturonic acid(GalA),glucose(Glu),galactose(Gal),and arabinose(Ara),although their ratios differ.The particle sizes of the four PGPs were mainly concentrated around 300 nm,and all demonstrate good solution stability.Compared with the OXA alone group,the OXA+PGP-CA and OXA+PGP-KA groups increased the lung tissue content of OXA by 36.93%and 35.12%,respectively.The OXA+PGP-KA group also significantly increased the expression level of Oct-2 protein(P<0.05),while there was no significant difference in the expression level of Pg-P protein.Conclusion:The physicochemical properties of PGP extracted by different processes are different.Among them,the combination of PGPCA and PGP-KA can significantly increase the lung tissue distribution of OXA,which would be related to the increase in Oct-2 expression in lung tissues.