河南新野产紫甘蓝花青素的提取工艺和调节双卵巢切除模型(OVX)小鼠糖耐量的研究  

Study on the Extraction Process of Anthocyanins from Brassica oleracea var.capitata rubra Cultivated in Xinye,Henan and Its Regulatory Effect on Glucose Tolerance in Ovariectomized(OVX)Mice

作　　者：周自亮 张艺菁 何叶 周少金 张亚丽 詹艳柯 曹红卫 张仲鸣 ZHOU Zi-liang;ZHANG Yi-jing;HE Ye(Agricultural Technology Application Center,Xichuan,Henan 474450;Zhang Zhongjing College of Chinese Medicine,Nanyang,Henan 473004)

机构地区：[1]河南省淅川县农业技术推广中心,河南淅川474450 [2]河南省张仲景国医国药学院,河南南阳473004 [3]河北邯郸天道酒业有限公司,河北邯郸056000

出　　处：《安徽农业科学》2023年第16期156-161,173,共7页Journal of Anhui Agricultural Sciences

基　　金：河南省自然科学基金项目(212300410376);南阳理工学院大学生科研基金项目(NG2019DKJ-070)。

摘　　要：[目的]探讨新野产紫甘蓝中花青素提取的最适条件,并揭示其对更年期模型动物糖耐量的调节作用。[方法]利用紫外-可见分光光度计和Greey公式直接计算提取液中花青素光密度和含量,探讨乙醇浓度、温度、提取时间和料液比等单因素对花青素提取的作用,并利用正交试验获得最佳提取条件;利用双卵巢切除模型小鼠(OVX)和腹腔葡萄糖耐量试验(IPGTT)研究紫甘蓝花青素对模型小鼠糖耐量的改善作用,利用定量PCR探讨相关机理。[结果]单因素试验发现,在酸化乙醇的条件下,乙醇浓度为40%、温度为55℃、提取时间为4 h、料液比为1∶15(g∶mL)时提取效果最好;正交试验表明,提取温度70℃、乙醇浓度60%、提取时间4 h、料液比1∶20为最佳提取条件,在此条件下,1 kg紫甘蓝中可得到花青素148.8 mg。花青素灌胃[100 mg/(kg·d),0.5 mL]处理OVX小鼠70 d后,相较假手术对照组,OVX手术导致小鼠糖耐量显著降低,曲线下面积(AUC)增加37.2%(P<0.05);普通甘蓝对糖耐受的调节并不显著,紫甘蓝花青素则可使OVX动物糖耐量的AUC降低30.5%,与假手术组的基本相当。定量PCR研究发现,相对假手术组(Shm+H_(2)O),OVX手术(OVX+H_(2)O)小鼠肝脏葡萄糖-6-磷酸酶(G6Pase)的表达水平升高了(5.22±1.02)倍,OVX+花青素组(OVX+BOR)和OVX+0.02 mg/(kg·d)补佳乐(OVX+E_(2))的表达量分别升高了(2.40±0.34)和(1.82±0.18)倍,与OVX+H_(2)O和OVX+BO组相比,OVX+E_(2)组、OVX+BOR组均显著降低了肝脏中G6Pase水平。[结论]紫甘蓝是天然花青素的重要来源,其可通过降低OVX小鼠肝脏中G6Pase的表达改善糖耐量,对缓解更年期高血糖等代谢综合征有一定的价值。[Objective]To explore the optimal conditions for the extraction of anthocyanidin from Brassica oleracea var.capitata rubra and reveal its regulation on the glucose tolerance of climacteric model animals.[Method]The optical density and content of anthocyanidin in the extraction solution were directly calculated by using the ultraviolet visible spectrophotometer and the Greey formula,and the effects of ethanol concentration,temperature,extraction time and solid-liquid ratio on the extraction of anthocyanidin were discussed,and the optimal extraction conditions were obtained by orthogonal test.The improvement effect of anthocyanidin from Brassica oleracea var.capitata rubra on glucose tolerance in ovariectomized mice(OVX)and intraperitoneal glucose tolerance test(IPGTT)was studied,and the related mechanism was explored by quantitative PCR.[Result]The single factor experiment found that under the conditions of acidified ethanol,the extraction effect was best when the ethanol concentration was 40%,the temperature was 55℃,the extraction time was 4 hours,and the solid-liquid ratio was 1∶15(g∶mL).Orthogonal test showed that the optimum extraction conditions were extraction temperature of 70℃,ethanol concentration of 60%,extraction time of 4 h,solid-liquid ratio of 1∶20.Under these conditions,148.8 mg anthocyanidin could be obtained from 1 kg Brassica oleracea var.capitata rubra.After the OVX mice were treated with anthocyanidin[100 mg/(kg·d),0.5 mL]by gavage for 70 days,compared with the Sham surgery control group,the glucose tolerance of the mice was significantly reduced and the area under the curve(AUC)was increased by 37.2%(P0.05);the regulation of sugar tolerance of ordinary cabbage was not significant,while anthocyanidin from Brassica oleracea var.capitata rubra could reduce the AUC of sugar tolerance of OVX animals by 30.5%,basically equivalent to that of the Sham surgery group.The quantitative PCR study found that compared with the Sham surgery group(Shm+H_2O),the expression level of glucose-6-phosphat

关 键 词：紫甘蓝 花青素 提取工艺 糖耐量 双卵巢切除模型 

分 类 号：R28[医药卫生—中药学]