人参-丁香醇提物抑制高山被孢霉脂质合成的活性评价  

Activity Evaluation of Alcohol Extract of Ginseng-Clove in Inhibiting Lipid Synthesis of Mortierella alpina

作　　者：倪伟锋 赵大庆 倪以宇 白竹林 王思明 NI Weifeng;ZHAO Daqing;NI Yiyu;BAI Zhulin;WANG Siming(Jilin Ginseng Academy,Changchun University of Traditional Chinese Medicine,Changchun 130000,China)

机构地区：[1]长春中医药大学吉林省人参科学研究院,吉林长春130000

出　　处：《食品工业科技》2022年第21期388-395,共8页Science and Technology of Food Industry

基　　金：人参产业关键技术研究及大健康产品开发,“十三五”国家重点研发计划(2017YFC1702100);国家级大学生创新创业训练计划项目(202010199081)。

摘　　要：探究人参-丁香醇提物抑制脂质合成并对其机制进行初步研究。通过建立丰产脂肪酸的高山被孢霉(Mortierella alpina,MA)脂质筛选模型,利用2,3,5-氯化三苯基四氮唑(TTC)筛选出脂质合成抑制最优的人参丁香药对用药比例和提取溶媒,测定MA内甘油三酯含量、电镜观察其形态并进行脂肪酸气质(GC-MS)分析,通过RT-PCR法测定乙酰辅酶A羧化酶(ACC)、ATP-柠檬酸裂解酶(ACLY)、脂肪酸合酶(FAS)和苹果酸酶(ME)基因的mRNA表达并对相关酶活力进行检测,对3T3-L1前脂肪细胞采用MTT法测定其增殖活力同时采用油红O染色法测定细胞分化中脂质积累的能力并加以论证。结果显示,人参丁香药对抑制脂质合成的最优比例和提取溶媒是人参-丁香质量比为1:2的100%乙醇提取物(alcohol extract of genseng and clove,AEGC),AEGC在0~30μg/mL浓度内对3T3-L1前脂肪细胞的增殖、分化无细胞毒性,能抑制脂肪细胞分化过程中甘油三酯的形成和积累;RT-PCR结果显示,较空白组AEGC干预后,MA内ACC、ACLY、FAS和ME的m RNA水平显著下调(P<0.05),且ME、葡萄糖-6-磷酸脱氢酶(G-6-P)、ACLY和FAS的酶活力受到显著抑制(P<0.05)。筛选所得的AEGC能抑制高山被孢霉和3T3-L1前脂肪细胞的脂质合成。The inhibitory of lipid synthesis by the alcohol extract of ginseng-clove was explored and its mechanism was preliminarily studied. A lipid screening model of Mortierella alpina(MA) which is rich in fatty acids was established, and2,3,5-triphenyltetrazolium chloride(TTC) was used to screen out the optimal proportion and extraction solvent of ginsengclove for inhibiting lipid synthesis. Then the content of triglyceride in MA was measured, its morphology was observed by electron microscope, and fatty acid was analyzed by GC-MS. The mRNA expressions of acetyl-CoA carboxylase(ACC),ATP-citric acid lyase(ACLY), fatty acid synthase(FAS) and malic enzyme(ME) were determined by RT-PCR, and the activities of related enzymes were detected. 3T3-L1 preadipocytes were demonstrated by MTT to determine proliferation viability and oil red O staining to determine the ability of lipid accumulation in cell differentiation. The results showed that the optimal ratio and extraction medium for the inhibition of lipid synthesis was the 100% alcohol extract of ginseng and clove(AEGC) with a 1:2 mass ratio of ginseng and clove. The extract was not cytotoxic to the proliferation and differentiation of 3T3-L1 preadipocytes at concentrations ranging from 0~30 μg/mL, and inhibited the formation and accumulation of triglycerides during the differentiation of adipocytes. The RT-PCR showed that the mRNA levels of ACC, ACLY, FAS and ME in MA were significantly downregulated(P<0.05) and the enzymatic activities of ME, glucose-6-phosphate dehydrogenase(G-6-P), ACLY and FAS were significantly inhibited(P<0.05) compared with the blank group after AEGC intervention. The AEGC obtained from the screening inhibited lipid synthesis in MA and 3T3-L1.

关 键 词：高山被孢霉 脂肪细胞 脂质合成 人参 丁香 

分 类 号：TS201.4[轻工技术与工程—食品科学]