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 HaCaT 细胞 损伤模型，观察 ACSP 对 HaCaT 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（mTOR） mRNA 表达水平和 p21、p53、p-p53 蛋白表达的影响，探讨 ACSP 对 MGO 诱发 HaCaT 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（mL∶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 HaCaT 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、 CAT mRNA 表达（P＜0.05，P＜0.01）；降低 MDA 酶活力（P＜0.05），下调 p53、p21、PI3K、Akt、mTOR mRNA 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。

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 HaCaT 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 HaCaT 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 mRNA expression levels of p53，p21，SOD，CAT，phosphatidylinositol 3-kinase （PI3K），protein kinase B （Akt）， mammalian target of rapamycin （mTOR），and the protein expressions of p21，p53 and p-p53 in HaCaT cell injury model were observed to invetigate the protective effect and mechanism of action of ACSP on MGO-induced HaCaT 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 （mL∶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 HaCaT cells （P＜0.05，P＜0.01），and up-regulate the mRNA expressions of SOD and CAT （P＜0.05，P＜0.01）. MDA enzyme activity was decreased （P＜0.05），mRNA expressions of p53，p21， PI3K， Akt and mTOR 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 HaCaT cell damage，and its mechanism is related to the regulation of p53/p21 signaling pathway.

R285.5

国家中医药管理局中医药国际合作专项项目（XDZYJZC-001）；广东省中医院中医药科学技术研究专项项目（YN2023MS44）