藏药黄花香薷联合阿苯达唑治疗大鼠继发性多房棘球蚴感染的疗效研究

doi:10.12140/j.issn.1000-7423.2020.06.003

摘要/Abstract

摘要：

目的观察藏药黄花香薷和阿苯达唑单用及联用治疗大鼠继发性原位多房棘球蚴感染的疗效,探讨藏药联合西药治疗多房棘球蚴感染的新途径。方法 SD雄性大鼠开腹暴露肝脏,注入棘球蚴原头节混悬液0.2 ml（含2 × 10³个原头节）,建立大鼠继发性多房棘球蚴感染模型。40只感染模型大鼠随机均分为50 mg/(kg·d)阿苯达唑组、50 mg/(kg·d)黄花香薷组、25 mg/(kg·d)阿苯达唑 + 25 mg/(kg·d)黄花香薷联合用药组和感染对照组,另取10只健康SD大鼠为健康对照组。3个用药组大鼠每天灌胃给药1次,4周为1疗程,间隔5 d,共2个疗程;感染对照组和健康对照组小鼠均给予等体积生理盐水。治疗2个疗程后,各组大鼠称重,取肝脏测量病灶直径大小;取脾脏称重,计算脾脏指数;取肝脏和脾脏病灶组织,切片后行HE染色,于光学显微镜下观察组织病理变化,肝脏病灶组织另于透射电镜下进行超微结构观察。结果阿苯达唑组、黄花香薷组和联合用药组大鼠肝脏病灶直径分别为（0.72 ± 0.06）、（0.79 ± 0.10）、（0.62 ± 0.05）cm,均小于感染对照组的（1.10 ± 0.11）cm（P < 0.01）,联合用药组的病灶直径小于阿苯达唑组和黄花香薷组,差异有统计学意义（P < 0.01）。联合用药组脾脏指数为5.28 ± 0.59,大于阿苯达唑组（3.35 ± 0.43）和黄花香薷组（3.56 ± 0.40）（P < 0.01）。HE染色观察结果显示,感染组和3个用药组的大鼠脾脏组织中白髓增殖明显,可见大量异形巨核细胞;3个用药组大鼠肝脏病灶组织均显示囊壁塌陷,其内原头节显著减少。黄花香薷组可见明显淋巴细胞浸润,联合用药组病灶组织坏死和玻璃样变明显。肝脏病灶组织超微结构观察显示,黄花香薷和阿苯达唑单用及联用治疗均能导致多房棘球蚴生发膜细胞线粒体结构破坏;联合治疗组肝棘球蚴囊壁结构疏松,生发层皮层区微绒毛变粗变短或者消失;黄花香薷组嗜酸粒细胞和巨噬细胞增加明显。结论黄花香薷和阿苯达唑联合治疗大鼠继发性多房棘球蚴感染有良好的效果。

关键词: 多房棘球蚴感染, 藏药, 黄花香薷

Abstract:

Objective To investigate the efficacy of the Tibetan medicine Elsholtzia eriostachya and albendazole, either alone or in combination, in treatment of secondary Echinococcus multiulocularis metacestode in rats, and to explore a new approach for treatment of the metacestode infection with Tibetan medicine combined with western medicine. Methods The SD male rats were laparotomized to expose the liver for injection with 0.2 ml of protoscoleces suspension (containing 2 × 10³ protoscoleces) to establish the rat model of secondary E. multiulocularis metacestode infection. The 40 model rats were divided into the albendazole treatment group [50 mg/(kg·d)], Elsholtzia eriostachya treatment group [50 mg/(kg·d)], albendazole [25 mg/(kg·d)]+Elsholtzia eriostachya [25 mg/(kg·d)] group, and the infection control group. Another 10 healthy SD rats were used as the healthy control group. Rats in the 3 treatment groups received two courses of treatment by gavage with once daily for consecutive 4 weeks, at an interval of 5 days between courses. Rats in the infection control group and healthy control group were given an equal volume of normal saline. After two courses of treatment, the rats body were weighed, the liver lesion diameter was measured, and the spleen index was calculated. The liver and spleen tissues in each group were obtained and sectioned for HE staining. Histopathological changes were observed under optical microscope and the ultrastructure of liver lesions was further observed under a transmission electron microscope. Results The diameters of rat liver lesions in the albendazole group, Elsholtzia eriostachya group and the combination group were (0.72 ± 0.06) cm, (0.79 ± 0.10) cm, and (0.62 ± 0.05) cm, respectively, all of which were smaller than the (1.10 ± 0.11) cm of the control group (P < 0.01). The diameter of the lesion in the combination group was significantly smaller than that of the albendazole group and the Elsholtzia eriostachya group (P < 0.01). The spleen index of the combined medication group was 5.28 ± 0.59, which was significantly higher than that of the albendazole group (3.35 ± 0.43) and the Elsholtzia eriostachya group (3.56 ± 0.40) (P < 0.01). Furthermore, the HE staining revealed obvious proliferation of white marrow in rat spleen tissues in the infection group and the three treatment groups, with the presence of a large number of heteromorphic megakaryocytes. The liver lesions in three treatment groups all showed collapse of the cyst wall and significant reduction of protoscoleces. Obvious lymphocyte infiltration was seen in the Elsholtzia eriostachya group, and necrosis and hyaline degeneration were obvious in the combined treatment group. Ultrastructural observations of liver tissues showed that treatment with Elsholtzia eriostachya and albendazole alone or in combination lead to the destruction of the mitochondrial structure of the germinal membrane cells of the metacestode. In the combination treatment group, the metacestode showed loose wall structure, and the microvilli of the germinal cortex became thicker, shorter or even disappeared,. eosinophils and macrophages increased significantly in the Elsholtzia eriostachya group. Conclusion The treatment with combination of Elsholtzia eriostachya and albendazole presented a good therapeutic effect on secondary E. multiulocularis metacestode infection in rats.

Key words: Echinococcus multilocularis protoscoleces infection, Tibetan medicine, Elsholtzia eriostachya

中图分类号:

R532.32

朱吉海, 曹得萍, 切羊让中, 刘珺, 赵珺, 刘燕. 藏药黄花香薷联合阿苯达唑治疗大鼠继发性多房棘球蚴感染的疗效研究[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(6): 688-694.

ZHU Ji-hai, CAO De-ping, QIE Yangrangzhong, LIU Jun, ZHAO Jun, LIU Yan. Effect of Elsholtzia eriostachya in combination with albendazole in treatment of secondary Echinococcus multilocularis metacestode infection in rats[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2020, 38(6): 688-694.

图/表 3

图1

图2

图3

参考文献 27

[1]	Cadavid Restrepo AM, Yang YR, McManus DP, et al. The landscape epidemiology of echinococcoses[J]. Infect Dis Poverty, 2016,5:13.
[2]	Deplazes P, Rinaldi L, Alvarez Rojas CA, et al. Global distribution of alveolar and cystic echinococcosis[J]. Adv Parasitol, 2017,95:315-493.
[3]	Romig T, Deplazes P, Jenkins D, et al. Ecology and life cycle patterns of Echinococcus species[M] //Thompson A. Echinococcus and Echinococcosis, Part A. Amsterdam: Elsevier, 2017: 213-314.
[4]	Pensel PE, Castro S, Allemandi D, et al. Enhanced chemoprophylactic and clinical efficacy of albendazole formulated as solid dispersions in experimental cystic echinococcosis[J]. Vet Parasitol, 2014,203(1/2):80-86.
[5]	Gao HJ, Chen B, Zhang HB, et al. Pharmacokinetics and liver targeting of lyophilized albendazole nanoliposomes in rats[J]. Chin J Hosp Pharm, 2017,37(8):702-706. (in Chinese)
[5]	( 高惠静, 陈蓓, 张海波, 等. 阿苯达唑纳米脂质体冻干粉在大鼠体内药动学及肝靶向研究[J]. 中国医院药学杂志, 2017,37(8):702-706.)
[6]	Wen H, Li HT. Advances in surgical operation and drug therapy for hepatic echinococcosis[J]. Chin Animal Health, 2017,19(7):29-32. (in Chinese)
[6]	( 温浩, 李海涛. 肝包虫病的外科手术及药物治疗进展[J]. 中国动物保健, 2017,19(7):29-32.)
[7]	Samuel F, Degarege A, Erko B. Efficacy and side effects of albendazole currently in use against Ascaris, Trichuris and hookworm among school children in Wondo Genet, southern Ethiopia[J]. Parasitol Int, 2014,63(2):450-455.
[8]	Wan MRQ. Protection of medicinal plant resources of Tibetan medicine in Qinghai[J]. J Clin Med Lit Electron, 2017,4(A2):20186-20187. (in Chinese)
[8]	( 完玛仁青. 浅谈青海藏药药用植物资源保护[J]. 临床医药文献电子杂志, 2017,4(A2):20186-20187.)
[9]	Dimaer DZPC. Jing Zhu Materia Medica[M]. Shanghai: Shanghai Science and Technology Press, 2012: 213. (in Chinese)
[9]	( 帝玛尔·丹增彭措. 晶珠本草[M]. 上海: 上海科学技术出版社, 2012: 213.)
[10]	Flora of China Editorial Committee of Chinese Academy of Science. The Flora of China[M]. Beijing: Science Press, 2014. (in Chinese)
[10]	( 中国科学院中国植物志编辑委员会. 中国植物志[M]. 北京: 科学出版社, 2004.)
[11]	Sun LP, Wang JR, Li XR, et al. Studies on the chemical constituents of Elsholtzia eriostachys(Elsholtzia eriostachys)ⅱ.isolation and identification of flavonoids constituents[J]. Chin Tradit Herb Drugs, 1997(11):646-648. (in Chinese)
[11]	( 孙丽萍, 王建人, 李秀荣, 等. 黄花香薷化学成分的研究 Ⅱ. 黄酮类成分的分离和鉴定[J]. 中草药, 1997(11):646-648.)
[12]	Xie P, Bai Y, Ci DJ, et al. Study on quality standard of the Tibetan Eriostachys elsholtzia[J]. Tibet Sci Technol, 2012(8):67-69. (in Chinese)
[12]	( 谢平, 白央, 次德吉, 等. 藏药材黄花香薷的质量标准研究[J]. 西藏科技, 2012(8):67-69.)
[13]	Liu XP, Jing XM. Research of chemical composition and biological activity of the essential oil from elsholtziaciliata[J]. J Heilongjiang August First Land Reclam Univ, 2018,30(3):35-39. (in Chinese)
[13]	( 刘香萍, 敬雪敏. 香薷精油化学成分及其生物活性研究[J]. 黑龙江八一农垦大学学报, 2018,30(3):35-39.)
[14]	Jiang S, Tang H, Xiao SG. Clinical application study of “Xiangru (Mosla herb)”[J]. Guid J Tradit Chin Med Pharmacol, 2015,21(9):95-97. (in Chinese)
[14]	( 江岁, 唐华, 肖深根. 香薷的临床应用研究[J]. 中医药导报, 2015,21(9):95-97.)
[15]	Li M, Miao MS. Characteristic analysis of the elsholtzia’s chemistry and pharmacology in clinical application[J]. Chin J Chin Med, 2015,30(4):578-579. (in Chinese)
[15]	( 李敏, 苗明三. 香薷的化学、药理与临床应用特点分析[J]. 中医学报, 2015,30(4):578-579.)
[16]	Dai XD, Ren YX, Liu X, et al. Transmission electron microscope investigation of Trichomonas vaginalis treated with Mosla chinensis Maxim[J]. J Trop Med, 2011,11(12):1379-1382. (in Chinese)
[16]	( 戴晓冬, 任一鑫, 刘欣, 等. 香薷体外抗阴道毛滴虫的透射电镜观察[J]. 热带医学杂志, 2011,11(12):1379-1382.)
[17]	Zheng LL, Cui Y, Qin YH, et al. Effect of Mosla chinensis Maxim on Trichomonas vaginalis in vitro[J]. J Dalian Med Univ, 2009,31(3):282-285. (in Chinese)
[17]	( 郑莉莉, 崔昱, 秦元华, 等. 中药香薷体外杀灭阴道毛滴虫效果的观察[J]. 大连医科大学学报, 2009,31(3):282-285.)
[18]	Liu HD, Wang HB, Fan HN, et al. Alveolar echinococcosis and immune evasion[J]. Chin J Parasitol Parasit Dis, 2018,36(6):655-660. (in Chinese)
[18]	( 刘寒冬, 王宏宾, 樊海宁, 等. 多房棘球蚴病的免疫逃避机制[J]. 中国寄生虫学与寄生虫病杂志, 2018,36(6):655-660.)
[19]	Hou XL, Li LH, Li L, et al. Changes in subsets and functional exhaustion of CD4⁺ T cells in spleens of mice infected with Echinococcus multilocularis [J]. Chin J Parasitol Parasit Dis Dec, 2020,38(5):611-618, 624. (in Chinese)
[19]	( 侯昕伶, 李玲慧, 李亮, 等. 多房棘球蚴感染小鼠脾CD4⁺ T细胞亚群及其功能耗竭的变化 [J]. 中国寄生虫学与寄生虫病杂志, 2020,38(5):611-618, 624.)
[20]	Mills SE. Practical histology for pathologists[M]. Xue DB, Chen J, Wang W (trans.). Beijing: Beijing Science and Technology Press, 2016: 23. (in Chinese)
[20]	( Mills SE. 病理医师实用组织学[M]. 薛德彬, 陈健, 王炜, 译. 北京: 北京科学技术出版社, 2016: 23.)
[21]	Qin FY, Song HT, Zhang X, et al. Research progress in the role of profilin in parasitic infection and immunity[J]. Chin J Parasitol Parasit Dis, 2019,37(6):709-712. (in Chinese)
[21]	( 秦飞宇, 宋海桐, 张昕, 等. 抑制蛋白在寄生虫感染与免疫中的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2019,37(6):709-712.)
[23]	Jiang JQ, E EDMT, Bao HE, et al. Correlation between eosinophils and immunity to parasitic infection[J]. Heilongjiang Animal Sci Vet Med, 2016(2):63-67. (in Chinese)
[23]	( 姜建强, 额尔敦木图, 包花尔, 等. 嗜酸性粒细胞与寄生虫感染免疫的相关性[J]. 黑龙江畜牧兽医, 2016(3):63-67.)
[24]	Li HW, Yang YR. Research progress in the mechanism of Echinococcus evasion[J]. Immunol J, 2010,26(11):1000-1002. (in Chinese)
[24]	( 李红卫, 杨玉荣. 棘球蚴逃避或抑制宿主免疫系统清除作用的机制研究进展[J]. 免疫学杂志, 2010,26(11):1000-1002.)
[25]	Huang ZK, Li JM. Macrophage polarization and its role in infectious diseases[J]. Int J Immunol, 2012,35(4):255-258, 281. (in Chinese)
[25]	( 黄自坤, 李俊明. 巨噬细胞极化及其在感染性疾病中的作用[J]. 国际免疫学杂志, 2012,35(4):255-258, 281.)
[26]	Yao Y, Xu J, Huang GX, et al. Research progress of Moslae Herba and predictive analysis on its Q-marker[J]. Chin Tradit Herb Drugs, 2020,51(10):2661-2670. (in Chinese)
[26]	( 姚奕, 许浚, 黄广欣, 等. 香薷的研究进展及其质量标志物预测分析[J]. 中草药, 2020,51(10):2661-2670.)
[27]	Yang S, Xu YQ, Xing YY, et al. Research advances on effects of plant-based flavonoids on immune and antioxidative functions in animals[J]. Chin J Animal Nutr, 2019,31(7):2958-2964. (in Chinese)
[27]	( 杨硕, 徐元庆, 邢媛媛, 等. 植物源黄酮类化合物对动物免疫和抗氧化功能影响的研究进展[J]. 动物营养学报, 2019,31(7):2958-2964.)