【摘要】 目的 优化骆驼刺种子多糖（ACSP）的提取工艺及探讨ACSP对丙酮醛（MGO）诱导HaCaT细胞损伤的保护作用及机制。方法 考察不同提取温度、提取时间、提取次数、液料比因素对ACSP提取量的影响，并在此基础上采用响应面试验法优化ACSP的提取条件，筛选出ACSP的最佳提取条件。采用MGO复制Ha Ca T细胞损伤模型，观察ACSP对Ha Ca T细胞损伤模型的细胞存活率、氧化相关指标[超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）、谷胱甘肽（GSH）]及p53、p21、SOD、CAT、磷脂酰肌醇3激酶（PI3K）、蛋白激酶B（Akt）、哺乳动物雷帕霉素靶蛋白（m TOR） m RNA表达水平和p21、p53、p-p53蛋白表达的影响，探讨ACSP对MGO诱发Ha CaT细胞损伤的保护效果及作用机制。结果 当提取温度为90℃、提取时间为120 min、提取次数为3次、液料比为25∶1（m L∶g）时，ACSP提取量最高，为43.25 mg·g^-1。细胞实验显示，ACSP能够增加MGO诱导Ha CaT细胞的细胞存活率、GSH含量及SOD、CAT的酶活力（P<0.05,P<0.01），上调SOD、CATm RNA表达（P<0.05,P<0.01）；降低MDA酶活力（P<0.05），下调p53、p21、PI3K、Akt、m TOR m RNA表达和p-p53/p53、p21蛋白表达（P<0.05,P<0.01）。结论 该研究开发了一种简便高效的ACSP提取工艺，该工艺得到的ACSP有抗氧化作用，能够改善MGO诱导的HaCaT细胞损伤，其作用机制与调控p53/p21信号通路有关。更多还原

【Abstract】 Objective To optimize the extraction process of Alhagi camelorum seed polysaccharides （ACSP） and to investigate the protective effect and mechanisms of ACSP on methylglyoxal （MGO）-induced damage in Ha Ca T cells injury.Methods The effects of different extraction temperatures,extraction time,extraction times and liquid-to-feed ratios on the extraction yield of ACSP were investigated,and the optimal extraction conditions of ACSP were screened out by using the response surface test method to optimize the extraction conditions of ACSP on this basis.A model of Ha Ca T cell injury was established by employing MGO.The effects of ACSP on the cell survival rate,oxidation-related indicators[superoxide dismutase （SOD）,catalase （CAT）,malondialdehyde （MDA） and glutathione （GSH）],the m RNA expression levels of p53,p21,SOD,CAT,phosphatidylinositol 3-kinase （PI3K）,protein kinase B （Akt）,mammalian target of rapamycin （m TOR）,and the protein expressions of p21,p53 and p-p53 in Ha Ca T cell injury model were observed to invetigate the protective effect and mechanism of action of ACSP on MGO-induced Ha Ca T cell injury.Results At the extraction temperature of 90℃，the extraction time was 120 minutes,the extraction times was three times,and the liquid-to-feed ratio was 25∶1 （m L∶g）,the extraction amount of ACSP was the highest,which was 43.25 mg·g^-1.Cell experiments showed that ACSP could increase the cell viability,GSH content,SOD and CAT enzyme activity of MGO-induced Ha Ca T cells （P<0.05,P<0.01）,and up-regulate the m RNA expressions of SOD and CAT （P<0.05,P<0.01）.MDA enzyme activity was decreased （P<0.05）,m RNA expressions of p53,p21,PI3K,Akt and m TOR and protein expressions of p-p53/p53 and p21 were down-regulated （P<0.05,P<0.01）.Conclusion In this study,a simple and efficient ACSP extraction process was developed.The ACSP obtained by this process has antioxidant effect and can improve MGO-induced Ha Ca T cell damage,and its mechanism is related to the regulation of p53/p21 signaling pathway.更多还原