吡喹酮治疗对日本血吸虫感染小鼠脾巨噬细胞增殖和炎症反应的抑制作用  

Praziquantel inhibits splenic macrophage proliferation and inflammatory reaction in mice infected with Schistosoma japonicum

作　　者：王伟[1] 赵成思 缪婷婷[1] 周春雷 张成诚 秦敏 邵天业 王勇[1] WANG Wei;ZHAO Cheng-si;MIAO Ting-ting;ZHOU Chun-lei;ZHANG Cheng-cheng;QIN Min;SHAO Tian-ye;WANG Yong(Department of Pathogen Biology,Nanjing Medical University,Nanjing 211166,China;Department of Pathologyy Children's Hospital Affiliated to Nanjing Medical Universityy Nanjing 210008,China)

机构地区：[1]南京医科大学病原生物学系,南京211166 [2]南京医科大学附属儿童医院病理科,南京210008

出　　处：《中国寄生虫学与寄生虫病杂志》2020年第3期263-270,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.81471573)。

摘　　要：目的探讨吡喹酮治疗对日本血吸虫感染小鼠脾巨噬细胞数量和功能的影响。方法取35只雌性C57BL/6J小鼠随机分为健康对照组、未感染吡喹酮组、感染组、感染吡喹酮治疗组、溶剂对照组、过继转移组和过继转移治疗组等7组,每组5只,后5组每鼠经腹部皮肤感染日本血吸虫尾蚴(14±2)条。感染后10周,溶剂对照组小鼠尾静脉注射无菌生理盐水,过继转移组和过继转移治疗组小鼠尾静脉注射(1~2)×10^6个GFP+Ly-6C+单核细胞。未感染吡喹酮组、感染吡喹酮治疗组和过继转移治疗组小鼠给予吡喹酮治疗,每隔12 h灌胃300 mg/kg;健康对照组、感染组、溶剂对照组和过继转移组小鼠每隔12 h灌胃1%羧甲基纤维素溶液。连续28 d后,取健康对照组、未感染吡喹酮组、感染组和感染吡喹酮治疗组小鼠脾组织,采用HE染色评价吡喹酮治疗对小鼠脾组织损伤的影响;流式细胞术检测小鼠脾组织Ly-6C+巨噬细胞的数量和Ly-6C+巨噬细胞胞内Ki-67的表达,评价吡喹酮对其增殖的影响;实时荧光定量PCR(qRT-PCR)检测小鼠脾巨噬细胞炎症因子TNF-α、IL-1β的表达。流式细胞术检测溶剂对照组、过继转移组和过继转移治疗组小鼠脾中GFP+Ly-6C+巨噬细胞百分率,评价吡喹酮对其募集的影响。另取感染后10周小鼠脾贴壁巨噬细胞进行体外实验,吡喹酮处理组用20μg/ml吡喹酮处理24 h,溶剂对照组用二甲基亚砜处理,流式细胞术检测两组贴壁细胞中Ly-6C+巨噬细胞的百分率。结果外观及HE染色结果显示,健康对照组和未感染吡喹酮组小鼠脾脏实质内结构分明,蓝色白髓、红色红髓结构清楚,脾细胞排列规则,未出现脾脏病理损伤;感染组小鼠脾脏肿大,组织学结构紊乱,红髓、白髓分布不清,脾窦可见部分充血,细胞排列不规则;感染吡喹酮治疗组小鼠脾肿大的表现有所缓解,实质结构得以修复,红髓和白髓重新形成,脾细胞分布较规�Objective To explore the effect of praziquantel treatment on the number and function of splenic macrophages in mice infected with Schistosoma japonicum.Methods Thirty-five female C57 BL/6 J mice were randomly divided into healthy control group,uninfected plus praziquantel-treatment group,infected group,infected plus praziquantel-treatment group,solvent control group,adoptive transfer group and adoptive transfer treatment group(5 mice each).Mice in the latter five groups were each infected with 14±2 cercariae of S.japonicum through abdominal skin.Ten weeks after infection,the mice in the solvent control group were injected with sterile saline through tail vein,and the mice in the adoptive transfer group and the adoptive transfer treatment group were injected with(1-2)×10^6 green fluorescent protein(GFP)+Ly-6C+monocytes through tail vein.The mice in the uninfected plus praziquantel-treatment group,the infected plus praziquantel-treatment group and the adoptive transfer treatment group were given praziquantel treatment(300 mg/kg every 12 h)for consecutive 28 days.The mice in healthy control group,infected group,solvent control group and adoptive transfer group were gavaged with 1%carboxymethyl cellu-lose solution every 12 hours for consecutive 28 days.Then spleen tissues from each group of mice were taken to eval-uate the effect of praziquantel treatment on the spleen tissue damage by HE staining.To evaluate the effect of praz-iquantel on the proliferation of macrophages,the number of Ly-6C+macrophages and the expression of Ki-67 within Ly-6C+macrophages in the spleen were assessed by flow cytometry.Real-time fluorescence quantitative PCR(qRTPCR)was performed to detect the expression of inflammatory factors TNF-αand IL-1βin mouse splenic macrophages.Flow cytometry was used to calculate the proportions of GFP+Ly-6C+macrophages in the spleen in order to evaluate the effect of praziquantel on their recruitment.Adherent macrophages from spleens of mice 10 weeks post-infection were used for in vitro experiment.The ma

关 键 词：吡喹酮 巨噬细胞 日本血吸虫 脾肿大 炎症 

分 类 号：R532.21[医药卫生—内科学]