菟丝子粗多糖结构表征及其活性研究

Structural characterization and activity study of crude polysaccharide from Cuscuta chinensis

关键词:  菟丝子粗多糖  结构表征  抗氧化  美白

Key words:  Cuscuta chinensis crude polysaccharide  structural characterization  antioxidant  whitening

摘要:

[目的] 通过水提醇沉法制备菟丝子粗多糖，对其结构进行表征，探讨其抗氧化与美白能力。[方法] 采用水提醇沉法获得菟丝子粗多糖，采用苯酚-硫酸法、3，5-二硝基水杨酸比色法、离子色谱仪法、尺寸排阻色谱-多角度激光光散射法检测菟丝子粗多糖的多糖含量、单糖组成及分子量；采用体外1，1-二苯基-2-三硝基苯肼（DPPH）、2，2’-联氮-双（3-乙基苯并噻唑啉-6-磺酸）阳离子（ABTS+）自由基清除和双萤光素酶报告系统测定其抗氧化能力，并通过斑马鱼模型、黑素细胞刺激素（α-MSH）诱导B16细胞生成黑色素模型测定菟丝子粗多糖的美白功效。[结果] 菟丝子粗多糖提取率为11.04%，多糖含量为56.03%，重均分子量为74 565 g/mol，主要由甘露糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖组成。菟丝子粗多糖可以提高对DPPH、ABTS+自由基清除能力和抗氧化反应元件（ARE）的转录活性；减少斑马鱼体内黑色素生成并抑制B16细胞中黑色素含量及酪氨酸酶活性。[结论] 菟丝子粗多糖具有一定的抗氧化及美白能力，为天然活性成分开发应用于保健品和化妆品提供备选。

Abstract:

[Objective] To prepare the crude polysaccharide from Cuscuta chinensis（CCCP） by water extraction and alcohol precipitation，characterize its structure，and investigate its antioxidant and whitening activities. [Methods] CCCP was obtained by water extraction and alcohol precipitation. The polysaccharide content was determined by the phenol-sulfuric acid method. The monosaccharide composition was analyzed using high-performance anion-exchange chromatography with pulsed amperometric detection（HPAEC-PAD，referred to as ion chromatography in the text）. The molecular weight was determined by size-exclusion chromatography coupled with multi-angle laser light scattering（SEC-MALLS）. Antioxidant capacity was evaluated in vitro through 1，1-diphenyl-2-picryl-hydrazyl radical（DPPH） and 2，2’-Azinobis-（3-ethylbenzthiazoline-6-sulphonate）（ABTS⁺） radical scavenging assays and a dual-luciferase reporter system measuring antioxidant response element（ARE） transcriptional activity. Whitening efficacy was assessed using a zebrafish model and an α-melanocyte-stimulating hormone（α-MSH）-induced melanogenesis model in B16 cells. [Results] The extraction yield of CCCP was 11.04%，with a polysaccharide content of 56.03%. The weight-average molecular weight was 74 565 g/mol. CCCP was primarily composed of mannose，galacturonic acid，glucose，galactose，and arabinose. CCCP enhanced the scavenging capacity against DPPH and ABTS⁺ radicals and increased ARE transcriptional activity. Furthermore，CCCP reduced melanin synthesis in zebrafish and inhibited melanin content and tyrosinase activity in α-MSH-stimulated B16 cells. [Conclusion] CCCP possesses notable antioxidant and whitening activities，providing a candidate natural active ingredient for potential applications in health products and cosmetics.