摘要
采用硅胶柱层析法从多脂鳞伞(Pholiota adiposa)子实体中分离纯化出化合物F1-3,并通过电子轰击质谱及核磁方法鉴定其化学结构,确定该化合物为麦角甾-4,6,8(14),22-四烯-3-酮(ETO)。应用CCK8法测定ETO对HepG2、MCF7、He La及A549细胞增殖抑制率。在H22荷瘤小鼠体内进行ETO抗肿瘤实验,将小鼠随机分为模型组、阳性组(环磷酰胺CTX,0.1×10^(-3) mmol·kg^(-1))及ETO组(低、中、高剂量分别为0.025、0.05、0.1 mmol·kg^(-1)),计算脏器指数和抑瘤率;采用ELISA法测定小鼠血清中IL-2、IFN-γ、TNF-α、IL-6和VEGF含量,采用HE染色法观察肿瘤组织细胞排序、细胞完整度和细胞核数量,采用TUNEL法检测肿瘤组织细胞凋亡程度,采用免疫组化法检测VEGF、Bcl-2和BAX蛋白表达水平。结果表明ETO对Hep G2、MCF-7和He La细胞增殖具有抑制作用,100μg·m L^(-1)ETO对HepG2细胞的抑制率高达91.04%;在H22荷瘤小鼠体内实验中,ETO高剂量组抑瘤率达72.06%,且对小鼠胸腺、脾脏指数具有明显恢复作用,表现出抑制肿瘤生长的作用;免疫组化实验表明模型组小鼠肿瘤组织细胞排列整齐紧密,生长状态良好,凋亡率较低;ETO组小鼠肿瘤组织细胞出现不同程度的变形、破裂及模糊不清和大面积凋亡现象;与模型组相比,ETO可提高小鼠血清中IFN-γ、IL-2及IL-6含量,降低VEGF含量,通过降低VEGF、Bcl-2及升高BAX蛋白表达水平表现出较好的抗肿瘤作用。
Using silica gel column chromatography,a compound named F1-3 was isolated from Pholiota adiposa fruiting bodies,and then determined to be ergosta-4,6,8(14),22-tetraen-3-one(ETO)by MS and NMR.ETO was then studied for its inhibitory effects on HepG2,MCF-7,HeLa,and A549 cancer cells by Cell Counting Kit-8(CCK8),and antitumor effect on H22 tumor-bearing mice in vivo.ICR mice were randomly divided into five groups as follows:model group,positive group(cyclophosphamide,0.1×10^(-3) mmol·kg^(-1)),low dose ETO group(LD,0.025 mmol·kg^(-1)),medium dose ETO group(MD,0.05 mmol·kg^(-1))and high dose ETO group(0.1 mmol·kg^(-1)).Gavage with saline was the blank control.Mice in different groups were measured for tumor suppression rate and organ indices after 15 consecutive days of treatment.The contents of IFN-γ,IL-2,IL-6,TNF-αand VEGF in mouse serum were determined by ELISA.Tumor cell arrangement,cell integrity,number of nuclei were observed by hematoxylin-eosin(HE)staining,and the degree of apoptosis in tumor tissues was determined by TUNEL assay.The expression levels of VEGF,Bcl-2 and BAX were determined by immunohistochemistry.The results showed that ETO inhibited the proliferation of HepG2,MCF-7 and HeLa cells in vitro,i.e.the inhibition rate of 100μg·mL^(-1) ETO on HepG-2 cells reached 91.04%.ETO inhibited tumor growth in H22 mice,and the tumor inhibition rate of HD group reached 72.06%,with thymus index and spleen index significantly improved.In the model group,the tumor cells were arranged neatly and tightly,the growth status was good,and the apoptosis rate was low.In contrast,the tumor cells of the ETO treated groups showed different degrees of deformation,rupture,blurring,and massive apoptosis.Compared with the model group,ETO increased the contents of IFN-γ,IL-2 and IL-6 in mouse serum,decreased VEGF level,and showed a good antitumor effect through decreasing the expression levels of VEGF,Bcl-2,and increasing the expression level of BAX.
作者
刘丁瑞
王晓岩
LIU Dingrui;WANG Xiaoyan(Changchun Science-Technology University,Academy of Medical,Changchun 130600,Jilin,China;College of Traditional Chinese Medicine,Jilin Agricultural Science and Technology University,Academy of Traditional Chinese Medicine,Jilin 132109,Jilin,China)
出处
《食用菌学报》
CSCD
北大核心
2023年第2期56-64,共9页
Acta Edulis Fungi
基金
吉林省科技厅项目(20220508074RC)
吉林省教育厅项目(JJKH20220397KJ)。
