黄腐酚对小鼠肝多房棘球蚴生长的抑制作用

doi:10.12140/j.issn.1000-7423.2021.03.003

中国寄生虫学与寄生虫病杂志 ›› 2021, Vol. 39 ›› Issue (3): 304-310.doi: 10.12140/j.issn.1000-7423.2021.03.003

黄腐酚对小鼠肝多房棘球蚴生长的抑制作用

谭小武¹^,³(), 俞晓凡¹, 姜慧娇¹, 刑稚坤¹, 陈雪玲², 吴向未¹^,^*()

1 石河子大学医学院第一附属医院普外科,石河子 832008
2 石河子大学医学院免疫教研室,石河子832002
3 玉屏侗族自治县人民医院普外科,玉屏 554000

收稿日期:2020-09-18 修回日期:2020-11-16 出版日期:2021-06-30 发布日期:2021-07-05
通讯作者: 吴向未
作者简介:谭小武（1985-）,男,硕士,主治医师,主要从事普通外科疾病基础研究。E-mail： 448094431@qq.com
基金资助:
国家自然科学基金(81760570);国家自然科学基金(81760371);兵团中青年科技创新领军人才计划项目(2018CB017);兵团重点领域科技公关项目(2019AB031)

Inhibitory effect of xanthohumol on the growth of Echinococcus multilocularis metacestode in the liver of mice

TAN Xiao-wu¹^,³(), YU Xiao-fan¹, JIANG Hui-jiao¹, XING Zhi-kun¹, CHEN Xue-ling², WU Xiang-wei¹^,^*()

1 Department of Hepatobiliary Surgery, the First Affiliated Hospital of Medical College of Shihezi University, Shihezi 832008, China
2 Department of Immunology, Medical College of Shihezi University, Shihezi 832002, China
3 Department of General Surgery, Yuping Dong Autonomous County People’s Hospital, Yuping 554000, China

Received:2020-09-18 Revised:2020-11-16 Online:2021-06-30 Published:2021-07-05
Contact: WU Xiang-wei
Supported by:
National Natural Science Foundation of China(81760570);National Natural Science Foundation of China(81760371);Corps Young and Middle-aged Science and Technology Innovation Leading Talent Program Project(2018CB017);Science and Technology Public Relations Projects in Key Fields(2019AB031)

摘要/Abstract

摘要：

目的探讨黄腐酚对小鼠肝多房棘球蚴生长的抑制作用。方法将30只肝脏感染多房棘球蚴后30 d的C57BL/6小鼠随机分为感染组、溶剂组、黄腐酚组,每组10只;另设健康对照组（10只）。黄腐酚组每周经腹腔注射黄腐酚10 mg/kg,溶剂组腹腔注射等体积二甲基亚砜（DMSO）,健康对照组和感染组不作任何处理。治疗60 d后安乐处死各组小鼠,称量各组小鼠肝湿重和多房棘球蚴囊湿重,计算囊湿重抑囊率。ELISA检测各组小鼠血清中血管内皮生长因子（VEGF）的含量。制备肝病理组织切片,苏木精-伊红染色法（HE）观察肝多房棘球蚴组织病理学改变,免疫组化法和蛋白质免疫印迹（Western blotting）检测小鼠肝多房棘球蚴组织中核因子κB P65（NF-κB P65）、VEGF和微血管密度-高度糖基化的I型跨膜蛋白（MVD-CD34）的蛋白表达水平,以甘油醛-3-磷酸脱氢酶（GAPDH）为内参。结果黄腐酚治疗60 d后,黄腐酚组小鼠囊湿重为（0.458 ± 0.068）g,低于感染组的（1.088 ± 0.274）g和溶剂组的（1.973 ± 0.213）g（均P < 0.05）;黄腐酚组囊抑制率为57.7%（63/109）,高于溶剂组的10.4%(11.5/109.8)和感染组的0（均 P < 0.05）。ELISA检测结果显示,黄腐酚组小鼠血清中VEGF为（45.24 ± 1.93）pg/ml,低于感染组的（58.00 ± 5.56）pg/ml和溶剂组的（57.85 ± 16.00）pg/ml,高于健康对照组的（40.20 ± 3.24）pg/ml（均 P < 0.05）。小鼠肝多房棘球蚴病灶组织病理学检查结果示,黄腐酚组炎性反应带窄,生发层紊乱。免疫组化检测结果示：黄腐酚组小鼠肝多房棘球蚴组织中NF-κB P65的评分为（2.65 ± 1.14）,低于感染组的（9.46 ± 2.60）和溶剂组的（9.20 ± 1.64）（均 P < 0.05）;黄腐酚组VEGF的评分为（1.80 ± 0.83）,低于感染组（7.20 ± 1.30）和溶剂组的（7.43 ± 2.96）（均 P < 0.05）;黄腐酚组MVD-CD34的评分为（16.40 ± 1.14）,低于感染组的（40.60 ± 1.14）和溶剂组的（40.83 ± 2.16）（均 P < 0.05）。Western blotting检测结果显示,黄腐酚组小鼠肝多房棘球蚴组织中VEGF/GAPDH比值为（0.76 ± 0.82）,低于感染组的（2.84 ± 0.33）和溶剂组的（2.47 ± 0.12）,高于健康对照组的（0.63 ± 0.02）（均 P < 0.05）,而感染组与溶剂组相比差异无统计学意义（ P > 0.05）;黄腐酚组中MVD-CD34/GAPDH比值为（0.40 ± 0.02）,低于感染组的（1.11 ± 0.07）和溶剂组的（1.02 ± 0.14）,高于健康对照组的（0.14 ± 0.02）（均 P < 0.05）,感染组与溶剂组差异无统计学意义（ P > 0.05）;黄腐酚组中NF-κB P65/GAPDH比值为（0.24 ± 0.03）,低于感染组的（0.61 ± 0.05）和溶剂组的（0.66 ± 0.13）,高于健康对照组的（0.02 ± 0.01）（均 P < 0.05）。结论黄腐酚治疗肝多房棘球蚴感染小鼠60 d后,对肝多房棘球蚴的生长和血管生成有一定的抑制作用。

关键词: 多房棘球蚴, 黄腐酚, 血管内皮生长因子, 微血管密度, 核因子-κB

Abstract:

Objective To explore the inhibitory effect of xanthohumol on the growth of hepatic Echinococcus multilocularis metacestode in mice.Methods Thirty C57BL/6 mice with liver infection of E. multilocularis metacestode 30 d post infection were randomly assigned into the infection group, solvent group, and xanthohumol group (10 mice in each group); a health control group of 10 mice was also set. Mice in the xanthohumol group received weekly intraperitoneal injection of xanthohumol (10 mg/kg), while those in the solvent group were intraperitoneally injected with an equal volume of dimethyl sulfoxide (DMSO). The mice in the health control group and infection group received no any treatment. After 60 days of treatment, the mice of each group were euthanized. The wet weights of liver and the multilocular hydatid cysts in each group were recorded, and the inhibition rate of cyst wet weight was calculated. ELISA was performed to examine the serum content of vascular endothelial growth factor (VEGF) in the mice in each group. The histopathological changes of hepatic multilocular hydatid cysts were observed by HE staining. Immunohistochemistry and Western blotting were used to detect the protein levels of nuclear factor kappa-B P65 (NF-κB P65), VEGF and microvessel density-highly glycosylated type I transmembrane glycoprotein (MVD-CD34) in the hepatic multilocular metacestode tissues in mice, with the glyceraldehyde-3-phosphate dehydrogenase（GAPDH） as internal control.Results After 60 days of xanthohumol treatment, the cyst wet weight in the xanthohumol group was (0.458 ± 0.068) g, which was lower than in the infection group (1.088 ± 0.274) g and the solvent group (1.973 ± 0.213) g (bothP < 0.05); the cyst inhibition rate of the xanthohumol group was 57.7% (63/109), which was higher than 10.4% (11.5/109.8) in the solvent group and 0 in the infection group (both P < 0.05). ELISA results showed that the serum level of VEGF in the xanthohumol group was (45.24 ± 1.93) pg/ml, which was lower than (58.00 ± 5.56) pg/ml in the infection group and (57.85 ± 16.00) pg/ml in the solvent group, but higher than (40.20 ± 3.24) pg/ml in the health control group (all P < 0.05). The liver pathological examination of the infected mice liver tissues showed that the xanthohumol group had narrow inflammatory reaction bands and disordered germinal layers. The results of immunohistochemistry showed that the score of NF-κB P65 in the multilocular metacestode tissue in the xanthohumol group was (2.65 ± 1.14), which was lower than that of the infection group (9.46 ± 2.60) and the solvent group (9.20 ± 1.64) (both P < 0.05). The VEGF score in the xanthohumol group (1.80 ± 0.83) was significantly lower than that of the infection group (7.20 ± 1.30) and the solvent group (7.43 ± 2.96) (both P < 0.05). The MVD-CD34 score in the xanthohumol group was (16.40 ± 1.14), which was significantly lower than that of the infection group (40.60 ± 1.14) and the solvent group (40.83 ± 2.16) (both P < 0.05). Western blotting results showed that the VEGF/GAPDH ratio in the xanthohumol group was (0.76 ± 0.82), which was lower than that of the infection group (2.84 ± 0.33) and the solvent group (2.47 ± 0.12), but was higher than that of the health control group (0.63 ± 0.02) (all P < 0.05), but there was no significant difference between the infection group and the solvent group ( P > 0.05). The MVD-CD34/GAPDH ratio in the xanthohumol group was (0.40 ± 0.02), which was lower than that of the infection group (1.11 ± 0.07) and the solvent group (1.02 ± 0.14), but was higher than that of the health control group (0.14 ± 0.02) (all P < 0.05). There was no significant difference between the infection group and the solvent group ( P > 0.05). The NF-κB P65/GAPDH ratio in the xanthohumol group was (0.24 ± 0.03), which was lower than that of the infection group (0.61 ± 0.05) and the solvent group (0.66 ± 0.13), but was higher than that of the health control group (0.02 ± 0.01) (all P < 0.05). Conclusion The growth and angiogenesis of hepatic multilocular metacestode in mice were inhibited after treatment with xanthohumol for 60 days.

Key words: Echinococcus metacestode, Xanthohumol, Vascular endothelial growth factor, Microvessel density, Nuclear factor kappa-B

中图分类号:

R383.33

谭小武, 俞晓凡, 姜慧娇, 刑稚坤, 陈雪玲, 吴向未. 黄腐酚对小鼠肝多房棘球蚴生长的抑制作用[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(3): 304-310.

TAN Xiao-wu, YU Xiao-fan, JIANG Hui-jiao, XING Zhi-kun, CHEN Xue-ling, WU Xiang-wei. Inhibitory effect of xanthohumol on the growth of Echinococcus multilocularis metacestode in the liver of mice[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2021, 39(3): 304-310.

图/表 4

图1

图2

图3

图4

参考文献 28

[1]	Kern P, Menezes da Silva A, Akhan O, et al. The echinococcoses: diagnosis, clinical management and burden of disease[J]. Adv Parasitol, 2017,96:259-369.
[2]	Organization WH. Guidelines for cystic and alveolar echinococcosis in human. WHO informal working group on echinococcosis[J]. Bull World Health Organ, 1996,74.
[3]	Torgerson PR, Schweiger A, Deplazes P, et al. Alveolar echinococcosis: from a deadly disease to a well-controlled infection. Relative survival and economic analysis in Switzerland over the last 35 years[J]. J Hepatol, 2008,49(1):72-77. doi: 10.1016/j.jhep.2008.03.023 pmid: 18485517
[4]	Li FQ, Yang M, Li B, et al. Initial clinical results of orthotopic liver transplantation for hepatic alveolar echinococcosis[J]. Liver Transpl, 2010,13(6):924-926. doi: 10.1002/(ISSN)1527-6473
[5]	Abulaihaiti M, Wu XW, Qiao L, et al. Efficacy of albendazole-chitosan microsphere-based treatment for alveolar echinococcosis in mice[J]. PLoS Negl Trop Dis, 2015,9(9):e0003950. doi: 10.1371/journal.pntd.0003950
[6]	Wei SS, Sun TL, Du J, et al. Xanthohumol, a prenylated flavonoid from Hops, exerts anticancer effects against gastric cancer in vitro[J]. Oncol Rep, 2018,40(6):3213-3222.
[7]	Scagliarini A, Mathey A, Aires V, et al. Xanthohumol, a prenylated flavonoid from hops, induces DNA damages in colorectal cancer cells and sensitizes SW480 cells to the SN38 chemotherapeutic agent[J]. Cells, 2020,9(4):932. doi: 10.3390/cells9040932
[8]	Saito K, Matsuo Y, Imafuji H, et al. Xanthohumol inhibits angiogenesis by suppressing nuclear factor-κB activation in pancreatic cancer[J]. Cancer Sci, 2018,109(1):132-140. doi: 10.1111/cas.2018.109.issue-1
[9]	Albini A, Dell'Eva R, Vené R, et al. Mechanisms of the antiangiogenic activity by the hop flavonoid xanthohumol: NF-kappa B and Akt as targets[J]. FASEB J, 2006,20(3):527-529. doi: 10.1096/fsb2.v20.3
[10]	Zhao X, Jiang K, Liang B, et al. Anticancer effect of xanthohumol induces growth inhibition and apoptosis of human liver cancer through NF-κB/p53-apoptosis signaling pathway[J]. Oncol Rep, 2016,35(2):669-675. doi: 10.3892/or.2015.4455
[11]	Sun G, Zheng C, Deng Z, et al. TRAF5 promotes the occurrence and development of colon cancer via the activation of PI3K/AKT/NF-κB signaling pathways[J]. J Biol Regul Homeost Agents, 2020,34(4):1257-1268.
[12]	Zhou Q, Yang XF, Han HH, et al. Correlation between angiogenesis and disease progression of hepatic Echinococcus multilocularis in C57BL/6 mice[J]. Chin J Parasitol Parasit Dis, 2019,37(3):302-310. (in Chinese)
	( 周青, 杨雄峰, 韩欢欢, 等. 小鼠肝多房棘球蚴中血管新生与病程发展的相关性研究[J]. 中国寄生虫学与寄生虫病杂志, 2019,37(3):302-310.)
[13]	Xu LZ, Yang WT. Criteria for the results of immunohistochemical reactions[J]. Chin Oncol, 1996(4):229-231. (in Chinese)
	( 许良中, 杨文涛. 免疫组织化学反应结果的判断标准[J]. 中国癌症杂志, 1996(4):229-231.)
[14]	Wen H, Turgan Aili Aji, Shao YM, et al. Research achievements and challenges for echinococcosis control[J]. Chin J Parasitol Parasit Dis, 2015,33(6):466-471. (in Chinese)
	( 温浩, 吐尔干艾力·阿吉, 邵英梅, 等. 棘球蚴病防治成就及面临的挑战[J]. 中国寄生虫学与寄生虫病杂志, 2015,33(6):466-471.)
[15]	Wu WP, Wang H, Wang Q, et al. A nationwide sampling survey on echinococcosis in China during 2012—2016[J]. Chin J Parasitol Parasit Dis, 2018,36(1):1-14.
	( 伍卫平, 王虎, 王谦, 等. 2012—2016年中国棘球蚴病抽样调查分析[J]. 中国寄生虫学与寄生虫病杂志, 2018,36(1):1-14.)
[16]	Cadavid Restrepo AM, Yang YR, McManus DP, et al. The landscape epidemiology of echinococcoses[J]. Infect Dis Poverty, 2016,5:13. doi: 10.1186/s40249-016-0109-x pmid: 26895758
[17]	Yuan M, Song X, Lv W, et al. Effect of anacardic acid against echinococcosis through inhibition of VEGF-induced angiogenesis[J]. Vet Res, 2019,50(1):3. doi: 10.1186/s13567-019-0621-7
[18]	Anciaux M, Demetter P, de Wind R, et al. Infiltrative tumour growth pattern correlates with poor outcome in oesophageal cancer[J]. BMJ Open Gastroenterol, 2020,7(1):e000431. doi: 10.1136/bmjgast-2020-000431
[19]	Dong F, Ruan S, Wang J, et al. M2 macrophage-induced lncRNA PCAT6 facilitates tumorigenesis and angiogenesis of triple-negative breast cancer through modulation of VEGFR2[J]. Cell Death Dis, 2020,11(9):728. doi: 10.1038/s41419-020-02926-8
[20]	Chen YZ, Wang DD, Peng H, et al. Epigenetically upregulated oncoprotein PLCE1 drives esophageal carcinoma angiogenesis and proliferation via activating the PI-PLCε-NF-κB signaling pathway and VEGF-C/Bcl-2 expression[J]. Mol Cancer, 2019,18(1):1. doi: 10.1186/s12943-018-0930-x
[21]	Ren XF, Chen CH, Luo YM, et al. lncRNA-PLACT1 sustains activation of NF-κB pathway through a positive feedback loop with IκBα/E2F1 axis in pancreatic cancer[J]. Mol Cancer, 2020,19(1):1-19. doi: 10.1186/s12943-019-1085-0
[22]	Patel M, Horgan PG, McMillan DC, et al. NF-κB pathways in the development and progression of colorectal cancer[J]. Transl Res, 2018,197:43-56. doi: 10.1016/j.trsl.2018.02.002
[23]	Colgate EC, Miranda CL, Stevens JF, et al. Xanthohumol, a prenylflavonoid derived from hops induces apoptosis and inhibits NF-kappaB activation in prostate epithelial cells[J]. Cancer Lett, 2007,246(1/2):201-209. doi: 10.1016/j.canlet.2006.02.015
[24]	Babykutty S, Priya PS, Nandini RJ, et al. Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP-2/9 expression via inhibiting ERK1/2 and reducing DNA-binding activity of NF-κB in colon cancer cells[J]. Mol Carcinog, 2012,51(6):475-490. doi: 10.1002/mc.20812
[25]	Matsuo Y, Sawai H, Ochi N, et al. Proteasome inhibitor MG132 inhibits angiogenesis in pancreatic cancer by blocking NF-κB activity[J]. Dig Dis Sci, 2010,55(4):1167-1176. doi: 10.1007/s10620-009-0814-4
[26]	Vanhoecke BW, Delporte F, Van Braeckel E, et al. A safety study of oral tangeretin and xanthohumol administration to laboratory mice[J]. In Vivo, 2005,19(1):103-107. pmid: 15796161
[27]	Dorn C, Bataille F, Gaebele E, et al. Xanthohumol feeding does not impair organ function and homoeostasis in mice[J]. Food Chem Toxicol, 2010,48(7):1890-1897. doi: 10.1016/j.fct.2010.04.030
[28]	Hussong R, Frank N, Knauft J, et al. A safety study of oral xanthohumol administration and its influence on fertility in Sprague Dawley rats[J]. Mol Nutr Food Res, 2005,49(9):861-867. pmid: 16092070

黄腐酚对小鼠肝多房棘球蚴生长的抑制作用

Inhibitory effect of xanthohumol on the growth of Echinococcus multilocularis metacestode in the liver of mice

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 28

相关文章 15

编辑推荐

Metrics

本文评价

[1]	曹得萍, 李嘉静, 宋梦微, 莫刚. 多房棘球蚴组织蛋白体外刺激肝星状细胞变化的实验观察[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 440-445.
[2]	朱爱娅, 王旭, 王江友, 王颖, 李杨, 宋珊, 耿燕, 兰子尧, 戴佳芮. 贵州省儿童多房棘球蚴病1例[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 520-523.
[3]	娆琬·托勒洪, 阿不都撒拉木·阿不力克木, 杨凌菲, 陈璐, 李钊, 贾芳, 宋涛. 超声表现诊断肝多房棘球蚴病的效果评价及因素分析[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 312-318.
[4]	蒉嫣, 薛垂召, 王旭, 刘白雪, 王莹, 王立英, 杨诗杰, 韩帅, 伍卫平, 肖宁. 2021年全国棘球蚴病防治进展[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(2): 142-148.
[5]	郭刚, 任远, 焦红杰, 武娟, 郭宝平, 齐文静, 李军, 张文宝. 腹腔感染棘球蚴微囊对小鼠肝脏多房棘球蚴感染及致病的影响[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(2): 156-162.
[6]	马慧, 种世桂, 陈根, 张伶慧, 秦俊梅, 赵玉敏. 多房棘球蚴病相关细胞信号通路的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(2): 223-227.
[7]	都涛, 胡春晖, 甘雪辉, 高攀, 张发斌. 砂生槐总生物碱水剂和片剂体外体内抗多房棘球蚴的效果[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(1): 15-22.
[8]	侯昕伶, 李德伟, 施阳, 王茂林, 孜比姑·肉素, 阿比旦·艾尼瓦尔, 郑旭然, 康雪娇, 王慧, 李静, 张传山. 多房棘球蚴感染小鼠腹腔ST2⁺ T细胞亚群功能及其免疫检查点分子表达变化[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(6): 708-716.
[9]	安秀青, 王苗苗, 周鸿乾, 孟凯, 蔡剑平, 刘光辉, 阿吉德, 杨金煜. 肝多房棘球蚴病微血管密度的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(6): 792-797.
[10]	毋德芳, 付永, 任宾, 张耀刚, 许晓磊, 庞明泉, 樊海宁. 青海人源两型棘球绦虫（棘球蚴）遗传多样性及分化时间研究[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(5): 610-615.
[11]	张婷婷, 杜秋沛, 郭新建, 张灵强, 王志鑫, 常正松, 赵乾, 王海久, 侯立朝. 肝多房棘球蚴病脉管侵犯的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(4): 516-523.
[12]	吴亮亮, 杨凌菲, 宋涛. 不同方式建立肝多房棘球蚴感染SD大鼠模型病灶的超声及病理表现[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(4): 549-552.
[13]	才仁, 任远, 米荣升, 郭刚, 齐文静, 张壮志, 郭宝平. 新疆新源县多房棘球蚴病病例特征及其病原基因多态性分析[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(2): 181-186.
[14]	张伶慧, 陈根, 种世桂, 沈辉, 马慧, 赵玉敏. 多房棘球蚴病中免疫细胞调控机制的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(1): 109-113.
[15]	阿卜杜艾尼·啊卜力孜, 排组拉沙拉依阿当, 塔来提·吐尔干, 张瑞青, 王慧, 张传山, 邵英梅, 吐尔干艾力·阿吉. 多房棘球蚴虫体蛋白介导NK细胞表面受体NKG2A对NK细胞功能的影响[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(1): 36-42.