姜黄素超分子包合物改善乙醇诱导LO2肝细胞DNA损伤的作用机制  

Mechanism of curcumin/cyclodextrin polymer inclusion complex improving DNA injury of LO2 liver cells induced by ethanol

作　　者：范土贵 曹笑皇 赵云涛 高加龙 刘颖[4] 刘海 钟赛意 秦小明 陈建平 FAN Tugui;CAO Xiaohuang;ZHAO Yuntao;GAO Jialong;LIU Ying;LIU Hai;ZHONG Saiyi;QIN Xiaoming;CHEN Jianping(College of Food Science and Technology,Guangdong Ocean University,Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety,Guangdong Province Engineering Laboratory for Marine Biological Products,Guangdong Provincial Engineering Technology Research Center of Seafood,Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution,Guangdong Provincial Modern Agricultural Science and Technology Innovation Center for Subtropical Fruit and Vegetable Processing,Zhanjiang 524088,China;Collaborative Innovation Center of Seafood Deep Processing(Dalian Polytechnic University)Dalian 116034,China;College of Chemistry and Food Science,Yulin Normal University,Yulin 537000,China;College of Costal Agricultural Sciences,Guangdong Ocean University,Zhanjiang 524088,China)

机构地区：[1]广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,广东省海洋生物制品工程实验室,广东省海洋食品工程技术研究中心,水产品深加工广东普通高等学校重点实验室,广东省亚热带果蔬加工科技创新中心,广东湛江524088 [2]海洋食品精深加工关键技术省部共建协同创新中心(大连工业大学),辽宁大连116034 [3]玉林师范学院化学与食品科学学院,广西玉林537000 [4]广东海洋大学滨海农业学院,广东湛江524088

出　　处：《食品与发酵工业》2022年第8期182-189,共8页Food and Fermentation Industries

基　　金：广东省自然科学基金面上项目(2020A1515010860,2021A1515012455);广东海洋大学创新强校项目(230419100);广东海洋大学“南海学者计划”项目(002029002009)。

摘　　要：该文主要考察了姜黄素超分子包合物(curcumin/β-cyclodextrin polymer inclusion complex,CUR/CDP)保护乙醇诱导LO2肝细胞损伤的分子机制。采用流式细胞仪检测CUR/CDP预处理对乙醇诱导细胞损伤后细胞周期分布的影响;Western blot检测CUR/CDP预处理对乙醇诱导细胞内蛋白表达水平变化的影响;活性氧(reactive oxygen species,ROS)试剂盒检测CUR/CDP预处理对乙醇诱导细胞内ROS水平的影响。结果发现,乙醇能显著提高细胞内G_(2)/M期细胞数目,从空白对照组的(8.83±0.46)%提高到(21.86±0.13)%。而经80μg/mL CUR/CDP预处理后乙醇诱导的细胞损伤显著降低,发现其细胞内G_(2)/M期细胞数目从乙醇处理组的(21.86±0.13)%降低到(4.76±0.36)%。进一步研究发现,CUR/CDP能够显著下调乙醇处理组中的p21、p-ATM、p-P53、γ-H2AX以及p-p38MAPK的蛋白表达量和上调cyclin B1和p-MDM2的蛋白表达量。此外,CUR/CDP作用于细胞后其细胞内ROS的荧光强度显著减弱,表明CUR/CDP能够抑制乙醇诱导细胞内ROS的产生。综上所述,CUR/CDP通过抑制细胞内ROS的产生下调细胞内与DNA损伤相关蛋白的表达,从而改善乙醇诱导的肝细胞损伤。To investigate the molecular mechanism of curcumin/β-cyclodextrin polymer inclusion complex(CUR/CDP) protecting injury of LO2 liver cells induced by ethanol, the effects of CUR/CDP pretreatment on ethanol-induced cell cycle distribution were detected by flow cytometry. Western blot was used to detect the effects of CUR/CDP pretreatment on the expression changes of intracellular proteins induced by ethanol. The effects of CUR/CDP pretreatment on the levels of intracellular reactive oxygen species induced by ethanol were detected by ROS assay kit. The results showed that ethanol significantly increased G_(2)/M phase population from(8.83±0.46)% to(21.86±0.13)% in the blank control group. 80 μg/mL CUR/CDP pretreatment resulted in a decrease in G2/M phase population from(21.86±0.13)% to(4.76±0.36)%. Further studies showed that CUR/CDP significantly down-regulated the expression levels of p21, p-ATM, p-P53, γ-H2 AX and p-p38 MAPK proteins, and up-regulated the expression levels of Cyclin B1 and p-MDM2 proteins in the ethanol treatment group. In addition, the fluorescence intensity of intracellular ROS was significantly reduced after CUR/CDP pretreatment, indicating that CUR/CDP could inhibit the production of intracellular ROS induced by ethanol. In conclusion, CUR/CDP can reduce the expressions of DNA damage-related proteins by inhibiting intracellular ROS production, thus improving ethanol-induced liver cells injury.

关 键 词：姜黄素超分子包合物 乙醇 LO2肝细胞 DNA损伤 保护机制 

分 类 号：TS201.4[轻工技术与工程—食品科学]