多房棘球蚴通过肝脏p38MAPK通路调控棘球蚴自身生长  

Echinococcus multilocularis metacestode regulates its own growth through hepatic p38MAPK signaling pathway

作　　者：徐刚 毛艺 李江[1] 张宏伟[1] 张永国[1] 吴向未[1] 彭心宇[1] 孙红[1] 杨婧[1] 陈骞[1] 张示杰[1] XU Gang;MAO Yi;LI Jiang;ZHANG Hongwei;ZHANG Yongguo;WU Xiangwei;PENG Xinyu;SUN Hong;YANG Jing;CHEN Qian;ZHANG Shijie(First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832000,Xinjiang,China;School of Medicine,Shihezi University,Shihezi 832000,Xinjiang,China)

机构地区：[1]石河子大学医学院第一附属医院,新疆石河子832000 [2]石河子大学医学院,新疆石河子832000

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第4期447-453,共7页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(8176120052,81860363)。

摘　　要：目的探讨多房棘球蚴对肝组织p38MAPK通路的影响及肝组织p38MAPK分子表达水平对多房棘球蚴的作用。方法从腹腔内保种的沙鼠取多房棘球蚴原头节,选取4~6周的C57BL/6J小鼠,随机分为感染组、未感染组和抑制组,每组10只。抑制组肝门静脉注射SB202190溶液(5 mg/kg,20%DMSO和80%生理盐水混合溶剂),未感染组和感染组仅注射溶剂,每周注射1次,共4周;感染组和抑制组小鼠开腹手术将500个原头节注射于肝脏被膜下,饲养8周。安乐处死后,取肝脏称重并计算肝重比(肝脏质量/体质量),取邻近(距离囊泡0.3 cm以内)及远离(距离囊泡1 cm以外)多房棘球蚴囊泡的肝组织,用石蜡包埋并切片,进行苏木精-伊红(HE)染色、过碘酸雪夫/糖原(PAS)染色、Masson染色并分析炎性带、肝脏纤维化范围。采用TRIzol法提取肝组织RNA,实时荧光定量PCR(qPCR)检测MAPK14基因的表达水平。取各组邻近多房棘球蚴囊泡肝组织,采用蛋白质免疫印迹(Western blotting)和肝脏组织切片免疫组织化学,检测p38MAPK及p-p38MAPK表达水平。结果未感染组肝重比为(6.49±0.19)%,感染组增加至(6.80±0.33)%(t=2.74,P<0.05);未感染组肝体积为(5.27±0.34)cm^(3),感染组增大至(5.80±0.49)cm^(3)(t=2.83,P<0.05)。HE染色结果显示,未感染组无炎性带,感染组和抑制组炎性带面积占比分别为(51.2±14.0)%和(23.8±9.8)%,抑制组与感染组相比肝组织坏死灶减少,炎性带减少(t=3.92,P<0.01)。PAS染色结果显示,未感染组、感染组、抑制组紫红色染色区域面积占比分别为(1.3±0.3)%、(7.4±1.8)%、(4.5±0.4)%,抑制组紫红色染色区域相比感染组减少(t=3.82,P<0.05)。Masson染色结果显示,未感染组无蓝染区域,感染组、抑制组蓝染区域占比分别为(34.9±4.1)%和(16.3±2.8)%,抑制组蓝染区域与感染组相比明显减少(t=9.16,P<0.01)。qPCR结果显示,与远离多房棘球蚴病灶的肝组织(1)相比,感染组邻近病灶肝组织MAPK14相对表Objective To investigate the influence of Echinococcus multilocularis on the p38MAPK signaling pathway in liver tissue and the affect of p38MAPK expression level on E.multilocularis metacestodes.Methods E.multilocularis protoscolices were collected from the abdominal cavity of preserved gerbils.C57BL/6J mice aged 4-6 weeks were randomly assigned into three groups as infected group,uninfected group,and inhibitory group,with 10 mice each group.The inhibitory group mice were injected via liver portal vein with SB202190 solution(5 mg/kg,in mixed volvent of 20%DMSO and 80%sterile saline),while the uninfected and infected groups received the solvent only,once a week,for total 4 weeks.Subsequently,both the infected and inhibitory groups were given by liver sub‑capsular injection with 500 protoscolices and raised for 8 weeks.After euthanasia,the mice livers were collected and weighed to calculate weight ratio(liver mass/body mass).Liver tissues adjacent to(within 0.3 cm)and distant from(1 cm away)the metacestode vesicles were collected.These liver samples were embedded with paraffin and sec‑tioned,and then stained with hematoxylin‑eosin(HE),periodic acid‑schiff(PAS),and Masson’s trichrome for analyzing the range of inflammatory bands and hepatic fibrosis.Utilizing the TRIzol method,RNA was extracted from the liver tissues to detect the expression levels of MAPK14 gene by quantitative PCR(qPCR).Subsequently,liver tissues adja‑cent to the vesicles from each group were examined for the expression levels of p38MAPK and p‑p38MAPK using Western blotting and immunohistochemical methods,respectively.Results The livermass/body mass weight ratio in the uninfected group was(6.49±0.19)%,while it was increased to(6.80±0.33)%in the infected group(t=2.74,P<0.05).The liver size in the uninfected group was(5.27±0.34)cm^(3),whereas it increased to(5.80±0.49)cm^(3) in the infected group(t=2.83,P<0.05).HE staining indicated the absence of inflammatory bands in the uninfected group,with the in‑fected and inhibitory groups

关 键 词：多房棘球蚴 多房棘球蚴病 P38MAPK p⁃p38MAPK 

分 类 号：R532.32[医药卫生—内科学]