收稿日期: 2020-09-18
修回日期: 2020-11-16
网络出版日期: 2021-07-05
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
Received date: 2020-09-18
Revised date: 2020-11-16
Online published: 2021-07-05
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)
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。
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 . DOI: 10.12140/j.issn.1000-7423.2021.03.003
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.
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