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。
收稿日期: 2026-02-06
修回日期: 2026-04-09
网络出版日期: 2026-06-10
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
Received date: 2026-02-06
Revised date: 2026-04-09
Online published: 2026-06-10
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)
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
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 . DOI: 10.12140/j.issn.1000-7423.2026.03.002
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
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