H3组蛋白甲基化转移酶抑制剂BIX-01294对细粒棘球蚴原头节体外杀伤效果评价

doi:10.12140/j.issn.1000-7423.2026.03.002

中国寄生虫学与寄生虫病杂志 ›› 2026, Vol. 44 ›› Issue (3): 320-327.doi: 10.12140/j.issn.1000-7423.2026.03.002

H3组蛋白甲基化转移酶抑制剂BIX-01294对细粒棘球蚴原头节体外杀伤效果评价

葛聪蕙¹(), 阿比旦·艾尼瓦尔¹, 肖雯颖¹, 唐娜¹, 孙胜¹, 王梦颖¹, 高毅¹, 阿依娜尔·吉恩斯¹, 胡秋¹, 李静¹, 王慧²^,¹, 张传山¹^,²^,^*()()

¹ 新疆医科大学基础医学院，新疆乌鲁木齐 830017
² 新疆医科大学第一附属医院临床医学研究院，新疆乌鲁木齐 830054

收稿日期:2026-02-06 修回日期:2026-04-09 出版日期:2026-06-30 发布日期:2026-06-10
通讯作者: *张传山（ORCID: 0000-0002-3241-2224），男，博士，研究员，博士生导师，从事寄生虫感染与免疫研究。E-mail：dashan0518@126.com
作者简介:葛聪蕙，女，硕士研究生，从事寄生虫感染与免疫研究。E-mail：g1580871476@163.com
第一联系人：作者贡献
葛聪蕙负责实验操作和论文撰写，张传山、王慧、阿比旦·艾尼瓦尔和李静负责论文修改和审校，肖雯颖、唐娜、孙胜、王梦颖参与实验操作并提供实验材料，高毅、阿依娜尔·吉恩斯、胡秋负责细粒棘球蚴的提取及培养。
基金资助:
国家重点研发计划(2023YFD1801200);国家重点研发计划(2023YFD1801202);新疆维吾尔自治区自然科学基金(2022D01D60);国家自然科学基金(82372279);国家自然科学基金(82160396);新疆维吾尔自治区天山创新团队计划(2024D14010);新疆维吾尔自治区青年科技拔尖人才项目(2022TSYCCX0106);新疆维吾尔自治区青年科技拔尖人才项目(2024TSYCCX0102);国家级科研创新人才及科研创新团队培育项目(XYD2024GR01)

In vitro activity of a histone H3 methyltransferase inhibitor BIX-01294 against Echinococcus granulosus

GE Conghui¹(), ABIDAN Ainiwaer¹, XIAO Wenying¹, TANG Na¹, SUN Sheng¹, WANG Mengying¹, GAO Yi¹, AYINAER Jiensi¹, HU Qiu¹, LI Jing¹, WANG Hui²^,¹, ZHANG Chuanshan¹^,²^,^*()()

¹ College of Basic Medical Sciences, Xinjiang Medical University, Urumqi 830017, Xinjiang, China
² Institute of Clinical Medical Research, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang, China

Received:2026-02-06 Revised:2026-04-09 Online:2026-06-30 Published:2026-06-10
Contact: *E-mail：dashan0518@126.com
Supported by:
National Key Research and Development Program(2023YFD1801200);National Key Research and Development Program(2023YFD1801202);Natural Science Foundation of Xinjiang Uygur Autonomous Region(2022D01D60);National Natural Science Foundation of China(82372279);National Natural Science Foundation of China(82160396);Xinjiang Uygur Autonomous Region Tianshan Innovation Team Program(2024D14010);Xinjiang Uygur Autonomous Region “Tianshan Talent” Cultivation Program for Young Top-notch Talents(2022TSYCCX0106);Xinjiang Uygur Autonomous Region “Tianshan Talent” Cultivation Program for Young Top-notch Talents(2024TSYCCX0102);National Program for Cultivating Scientific Research Innovation Talents and Teams(XYD2024GR01)

摘要/Abstract

摘要：

目的探讨使用H3组蛋白甲基化转移酶抑制剂BIX-01294干预对细粒棘球蚴原头节和中绦期囊泡活性的影响。方法采集当地屠宰场感染细粒棘球蚴的绵羊肝脏，无菌条件下分离获得原头节，使用不同浓度的BIX-01294（1、5、10、25、50 µmol/L）二甲基亚砜（DMSO）溶液进行体外干预，同时设置溶剂对照组（DMSO组），分别在干预后第3、6、9、12和15天收集虫体，亚甲基蓝染色观察原头节的活性情况，计算原头节存活率，绘制生存率曲线，采用梯形法计算存活率-曲线下面积（AUC）。收集25 µmol/L BIX-01294干预12 d后的原头节，通过苏木精-伊红（HE）染色观察干预后原头节变化。收集1、25 µmol/L BIX-01294干预1、6、9、12 d的原头节，通过Caspase-3/7荧光染色观察干预后原头节的凋亡情况。蛋白质免疫印迹（Western blotting）检测25 µmol/L BIX-01294干预12 d后原头节中H3K9me2的表达水平。利用25 µmol/L的BIX-01294干预体外培养的囊泡，在第1、3、6、9和12天通过显微镜观察BIX-01294对囊泡形态的影响，通过扫描电镜观察BIX-01294干预9 d后囊泡超微结构的变化。结果 25 µmol/L BIX-01294干预6 d时，原头节存活率为（36.45 ± 7.14）%，与干预3 d的（90.21 ± 3.72）%相比下降（t = 15.86，P < 0.01）；50 µmol/L BIX-01294干预3 d时，原头节存活率为（71.91 ± 7.92）%，与干预0 d的100%相比下降（t = 6.25，P < 0.05）；50 µmol/L BIX-01294干预6 d时，原头节存活率为（5.64 ± 1.48）%，与干预3 d时相比继续下降（t = 15.89，P < 0.01）。各浓度组BIX-01294的AUC数值随其浓度升高呈剂量依赖性降低，50 µmol/L组AUC最低（395.25%·d）。显微镜下50 µmol/L BIX-01294干预6 d后，原头节虫体皱缩，顶突和小钩脱落，钙颗粒减少，活动减弱；HE染色结果显示，25 µmol/L BIX-01294干预12 d后原头节结构松散、空洞且体积缩小呈不规则状，体壁变薄且不完整。25 µmol/L BIX-01294干预12 d时Caspase-3/7荧光表达强度最强。Western blotting结果显示，干预12 d后原头节中H3K9me2表达水平（0.53 ± 0.08）较DMSO组（1.00 ± 0.17）降低（t = 4.226，P < 0.05）。囊泡经25 µmol/L的BIX-01294干预后第6天开始出现不同程度的塌陷或皱缩，扫描电镜结果显示，干预后第9天囊泡超微结构发现明显改变，生发层和角质层结构分离，生发层结构紊乱。结论 BIX-01294对体外培养的细粒棘球蚴原头节和囊泡具有显著杀伤效果，且呈现时间和浓度依赖效应，并伴随H3K9me2表达降低及凋亡相关信号上调，表明BIX-01294具有潜在的抗棘球蚴应用价值。

关键词: 细粒棘球蚴, H3组蛋白甲基化转移酶, BIX-01294

Abstract:

Objective To investigate the effects of BIX-01294, a histone H3 methyltransferase inhibitor, on the viability of protoscoleces and metacestode vesicles of Echinococcus granulosus. Methods Liver samples were collected from E. granulosus-infected sheep in local slaughterhouses. E. granulosus protoscoleces were isolated under sterile conditions and exposed to BIX-01294 solutions (dissolved in DMSO) at concentrations of 1, 5, 10, 25, and 50 µmol/L, while a solvent control group (DMSO group) served as controls. Protoscoleces were collected at 3, 6, 9, 12, and 15 days post-exposure and stained with methylene blue to observe viability. The survival rate of protoscoleces was calculated, and a survival curve was plotted to estimate the area under the survival curve (AUC) was determined using the trapezoidal rule. Protoscoleces were collected following exposure to 25 µmol/L BIX-01294 for 12 days and stained with hematoxylin and eosin (HE) to observe post-exposure changes. Protoscoleces exposed to 1 and 25 µmol/L BIX-01294 for 1, 6, 9, and 12 days were collected and stained with Caspase-3/7 dyes to assess the apoptosis of protoscoleces. The expression of H3K9me2 was determined using Western blotting in protoscoleces exposed to 25 µmol/L BIX-01294 for 12 days. The morphology of vesicles was observed under a microscope following exposure to 25 µmol/L BIX-01294 in vitro for 1, 3, 6, 9, and 12 days, and ultrastructural changes in vesicles were examined using scanning electron microscopy following exposure to BIX-01294 for 9 days. Results The viability of protoscoleces was (36.45 ± 7.14)% on day 6 post-treatment with 25 µmol/L BIX-01294, which significantly reduced as compared with that [(90.21 ± 3.72)%] on day 3 post-treatment (t = 15.86, P < 0.01), and the viability of protoscoleces was (71.91 ± 7.92)% on day 3 post-treatment with 50 µmol/L BIX-01294, which significantly reduced as compared with that (100%) on day 0 post-treatment (t = 6.25, P < 0.01), while the viability was (5.64 ± 1.48)% on day 6 post-treatment with 50 µmol/L BIX-01294, which continued to reduce as compared with that on day 3 post-treatment (t = 15.89, P < 0.01). The AUC values of BIX-01294 decreased in a dose-dependent manner with increasing concentrations, with the lowest AUC seen in the 50 µmol/L BIX-01294 treatment group (395.25%·d). Microscopy displayed shrinkage of protoscoleces, shedding of rostellum and hooklets, reduction of calcium granules and decreased motility following treatment with 50 µmol/L BIX-01294 for 6 days, and HE staining revealed loose, cavitary and irregular protoscoleces with a reduced volume, and thinned and incomplete body wall following treatment with 25 µmol/L BIX-01294 for 12 days. The highest Caspase-3/7 fluorescence intensity was recorded in protoscoleces exposed to 25 µmol/L BIX-01294 for 12 days. Western blotting assay determined lower H3K9me2 expression in protoscoleces treated with BIX-01294 for 12 days than in the DMSO group [(0.53 ± 0.08) vs. (1.00 ± 0.17); t = 4.226, P < 0.05]. Collapse or shrinkage of vesicles was found since day 6 following treatment with 25 µmol/L BIX-01294. Scanning electron microscopy displayed remarkable ultrastructural alterations, separation of germinal layer and cuticle and disorganization of the germinal layer structure in vesicles 9 days post-treatment with BIX-01294. Conclusion BIX-01294 exhibits a remarkable in vitro activity against E. granulosus protoscoleces and vesicles in a time- and concentration-dependent manner, accompanied by reduced H3K9me2 expression and upregulation of apoptosis-related signals, indicating that BIX-01294 has a potential value against echinococcosis.

Key words: Echinococcus granulosus, Histone H3 methyltransferase, BIX-01294

中图分类号:

R383.33

葛聪蕙, 阿比旦·艾尼瓦尔, 肖雯颖, 唐娜, 孙胜, 王梦颖, 高毅, 阿依娜尔·吉恩斯, 胡秋, 李静, 王慧, 张传山. H3组蛋白甲基化转移酶抑制剂BIX-01294对细粒棘球蚴原头节体外杀伤效果评价[J]. 中国寄生虫学与寄生虫病杂志, 2026, 44(3): 320-327.

GE Conghui, ABIDAN Ainiwaer, XIAO Wenying, TANG Na, SUN Sheng, WANG Mengying, GAO Yi, AYINAER Jiensi, HU Qiu, LI Jing, WANG Hui, ZHANG Chuanshan. In vitro activity of a histone H3 methyltransferase inhibitor BIX-01294 against Echinococcus granulosus[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2026, 44(3): 320-327.

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组别 Group	第3天 Day 3	第6天 Day 6	第9天 Day 9	第12天 Day 12	第15天 Day 15
CON	95.41 ± 2.49	96.42 ± 2.23	99.29 ± 1.23	96.41 ± 2.26	97.08 ± 0.79
DMSO	96.97 ± 0.60	96.13 ± 0.95	98.46 ± 1.42	99.38 ± 0.74	97.94 ± 0.62
1 µmol/L BIX-01294	96.22 ± 1.82	96.48 ± 0.75	97.76 ± 0.53	98.32 ± 0.24	97.51 ± 0.38
5 µmol/L BIX-01294	96.36 ± 1.70	95.87 ± 1.06	97.11 ± 1.52	96.78 ± 2.79	94.37 ± 3.90
10 µmol/L BIX-01294	96.94 ± 0.61	94.37 ± 1.11	97.12 ± 0.35	96.00 ± 0.81	94.75 ± 1.60
25 µmol/L BIX-01294	90.21 ± 3.72	36.45 ± 7.14^b	5.50 ± 3.54^b	2.59 ± 0.85^b	0.25 ± 0.28^b
50 µmol/L BIX-01294	71.91 ± 7.92^a	5.64 ± 1.48^b	1.61 ± 1.73^b	0.00 ± 0.00^b	0.00 ± 0.00^b

H3组蛋白甲基化转移酶抑制剂BIX-01294对细粒棘球蚴原头节体外杀伤效果评价

In vitro activity of a histone H3 methyltransferase inhibitor BIX-01294 against Echinococcus granulosus

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