晚期日本血吸虫感染小鼠焦虑样行为学研究  

Study on anxiety-like behavior of mice with late-stage infection of Schistosoma japonicum

作　　者：汤宪时 季文翔 熊春蓉[1] 周永华[1] 许永良[1] 仝德胜[1] TANG Xian-shi;JI Wen-xiang;XIONG Chun-rong;ZHOU Yong-hua;XU Yong-liang;TONG De-sheng(Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention,Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology,Jiangsu Institute of Parasitic Diseases,Wuxi 214064,China)

机构地区：[1]国家卫生健康委员会寄生虫病预防与控制技术重点实验室,江苏省寄生虫与媒介控制技术重点实验室,江苏省血吸虫病防治研究所,无锡214064

出　　处：《中国寄生虫学与寄生虫病杂志》2022年第5期622-628,634,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：江南大学公共卫生研究中心项目(JUPH201838)。

摘　　要：症程度的关系。方法 取30只6周龄雄性C57BL/6J小鼠,随机分为2组:感染组小鼠(16只)经腹壁皮肤接触感染雌雄尾蚴20条/鼠,未感染组(14只)用不含尾蚴的去氯水处理,两组小鼠于感染后第8周灌服吡喹酮(300 mg/kg)。另设7只6周龄雄性APP/PS1双转基因小鼠(阿尔兹海默症模型小鼠)为阳性对照(转基因组),不作任何处理。3组小鼠于相同环境饲养至10月龄,观察各组小鼠在旷场试验(OFT)和高架十字迷宫试验(EPM)中的行为变化,以OFT的总路程、中央区路程、中央路程百分比、活动次数、活动时间、中央时间百分比、平均速度、中央平均速度和EPM中的开臂、闭臂和中央区的运动路程、停留时间和进入次数以及开臂内运动路程、停留时间和进入次数的百分比,评价各组小鼠的焦虑样行为。各组小鼠安乐死后,取肝脏,观察肝脏大体形态;肝组织切片经HE和Masson染色后,镜下观察肝脏病理变化和胶原纤维增生情况;测定肝脏胶原纤维特有的羟脯氨酸含量;应用荧光定量PCR检测肝组织α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1 (TGF-β1)、Ⅰ型胶原(ColⅠ)、 ColⅢmRNA相对转录水平。应用SPSS 21.0软件进行统计学分析。结果与未感染组比较,感染组小鼠肝脏颜色发暗,肝组织内有大量虫卵沉积,周围有炎性细胞浸润和胶原纤维增生。感染组小鼠的肝脏羟脯氨酸含量为(0.194±0.040)μg/mg,高于未感染组的(0.122±0.016)μg/mg和转基因组的(0.124±0.004)μg/mg (t=0.79、 0.86,均P <0.05)。感染组小鼠肝脏ColⅠ、 α-SMA、 TGF-β1、 ColⅢm RNA相对转录水平分别为3.55±1.76、 2.01±0.66、 2.21±0.64、 4.51±2.32,与未感染组比较差异无统计学意义(t=0.76、0.59、 0.17、 0.40,均P> 0.05)。感染组小鼠OFT中的活动时间(346.77±181.03) s和EPM中的开臂内运动路程(206.31±282.52) mm、开臂内运动路程百分比(7.51±9.90)%,以及开臂、闭臂和中央进入次数(1.32±1.51Objective To observe the behavioral characteristics of mice with late-stage schistosomiasis japonica and explore the relationship between late-stage infection of Schistosoma japonicum and anxiety. Methods Thirty 6-week-old male C57BL/6J mice were randomly divided into the infected group(n = 16) and the uninfected group(n = 14). The infected group was infected via abdominal skin contact with 20 cercariae of both sexes, and the uninfected group was treated with dechlorinated water without cercariae. Mice in both of groups were administered with praziquantel by gavage at 8 weeks post-infection. Additional 7 male APP/PS1 double transgenic mice(Alzheimer ’s disease model) at 6 weeks of age were used as positive control(transgenic group) without any treatment. Three groups of mice were reared in the same environment until they were 10 months old, to observe their behavioral changes in an open-field test(OFT) and elevated-plus maze test(EPM). In OFT, the total distance, the travelling distance in the central zone, the percentage of travelling distance in the centrol zone, the number of locomotor activity, the duration of locomotor activity, the time percentage spent in the centrol zone, the mean speed, the mean speed in the centrol zone were measured, and in EPM, the locomotor distance, retention time and the number of entry in the open arms/close arms/central zone, and the percentage of locomotor distance, retention time and number of entry in the open arms were recorded to evaluate the anxiety-like behavior of the three mice groups. The mice of all groups were euthanized, and the livers were collected to observe gross morphology. The liver tissue sections were stained using hematoxylin & eosin(HE) and Masson ’s trichrome stain for microscopic observation of pathological changes and collagen fiber hyperplasia. The content of liver hydroxyproline which is the specific amino acid constituent in collagen, was determined. The mRNA relative transcription levels of α-smooth muscle actin(α-SMA), transforming growth fact

关 键 词：晚期日本血吸虫感染 旷场试验 高架十字迷宫试验 焦虑 

分 类 号：R532.21[医药卫生—内科学]