顺铂短期给药诱发小鼠药源性虚证模型的评价研究  

Evaluation on drug-induced deficiency syndrome model by short-term administration of cisplatin in mice

作　　者：农淄心 邓琳[2] 贺健祥 张超峰 谢海纳 曹琳娜 张夜航 范晓燕 吴晓 方肇勤[1] 潘志强[1] NONG Zixin;DENG Lin;HE Jianxiang;ZHANG Chaofeng;XIE Haina;CAO Linna;ZHANG Yehang;FAN Xiaoyan;WU Xiao;FANG Zhaoqin;PAN Zhiqiang(School of Basic Medical Sciences,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学基础医学院,上海201203 [2]上海中医药大学附属曙光医院,上海201203

出　　处：《上海中医药大学学报》2021年第4期35-44,共10页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：上海市科委实验动物专项研究项目(19140905000);上海市进一步加快中医药事业发展三年行动计划项目[ZY(2018-2020)-CCCX-2001-01]。

摘　　要：目的:研究顺铂短期给药诱发小鼠药源性证候的属性及对类固醇激素合成功能的影响。方法:40只雄性ICR小鼠随机分为对照组及顺铂不同剂量(3、5、10 mg·kg^(-1)·d^(-1))组,每组10只。各药物干预组小鼠腹腔注射相应剂量的顺铂,连续5 d。分别于干预5 d后和停药5 d后,观察小鼠体征(体质量、摄食量、腋温、爪色泽),并进行行为学测试(抓力、旷场试验和转棒试验)。观察期后,取血清;分离肾脏、心脏、胸腺和脾脏,计算脏器指数;收集肾上腺和睾丸。ELISA检测血清皮质酮和睾酮含量;HE染色后光镜下观察肾上腺与睾丸组织形态学变化;PCR、Western blot分别检测肾上腺皮质类固醇激素合成酶和睾丸睾酮合成酶相关的mRNA、蛋白表达。结果:(1)干预5 d后,顺铂10 mg·kg^(-1)·d^(-1)组小鼠体质量、摄食量、腋温、水平运动及垂直运动较对照组均显著降低(P<0.05,P<0.01);顺铂3、5 mg·kg^(-1)·d^(-1)组小鼠腋温明显低于对照组,而爪色r值高于对照组(P<0.05,P<0.01)。停药5 d后,顺铂10 mg·kg^(-1)·d^(-1)组小鼠体质量、摄食量、在棒时间、抓力、腋温、爪色r值、水平运动及垂直运动较对照组均显著降低(P<0.05,P<0.01);顺铂3、5 mg·kg^(-1)·d^(-1)组小鼠垂直运动较对照组亦明显减少(P<0.01)。(2)与对照组相比,10 mg·kg^(-1)·d^(-1)顺铂作用显著抑制小鼠脾脏、胸腺、心脏,脏器指数明显降低(P<0.05,P<0.01),而肾脏指数显著升高(P<0.01);5 mg·kg^(-1)·d^(-1)顺铂作用亦抑制心脏(P<0.05)。(3)形态学观察显示,顺铂10 mg·kg^(-1)·d^(-1)组小鼠肾上腺皮质球状带细胞肿胀明显,顺铂5、10 mg·kg^(-1)·d^(-1)组小鼠可见各级生精细胞排列略紊乱,睾丸间质细胞收缩明显。(4)与对照组相比,顺铂10 mg·kg^(-1)·d^(-1)组小鼠血清皮质酮含量显著升高(P<0.01),血清睾酮含量无明显变化。(5)与对照组相比,顺铂3 mg·kg^(-1)·d^(-1)组小鼠肾上腺Star、Cyp21a1的mRNA表达Objective:To study the effects of short-term administration of cisplatin on syndrome and steroidogenesis in mice.Methods:Forty male ICR mice were randomly divided into the control group and cisplatin groups with different doses(3,5,10 mg·kg^(-1)·d^(-1)),10 mice in each group.The drug intervention groups were intraperitoneally injected with cisplatin at the corresponding dose for 5 days.After last administration and withdrawal for 5 days,the signs(body mass,food intake,axillary temperature and claw color)of mice were observed,and the behavioral tests(grip,open field test and rotarod test)were performed.After observation period,the serum was taken,the kidney,heart,thymus and spleen were separated and the organ index was calculated,and the adrenal gland and testis were collected.The serum levels of corticosterone and testosterone were detected by ELISA.The morphological changes of adrenal gland and testis were observed under light microscope after HE staining.The expressions of genes and proteins related to steroid hormone synthetase and testosterone synthetase were detected by PCR and Western blot respectively.Results:(1)After administration for 5 days,the body mass,food intake,axillary temperature,horizontal movement and vertical movement in the cisplatin 10 mg·kg^(-1)·d^(-1) group were significantly decreased compared with those in the control group(P<0.05,P<0.01);the axillary temperature of the cisplatin 3 and 5 mg·kg^(-1)·d^(-1) groups was significantly lower than that of the control group,while the r value of claw color was higher than that of the control group(P<0.05,P<0.01).After withdrawal for 5 days,the body mass,food intake,stay time,grip,axillary temperature,r value of claw color,horizontal movement and vertical movement in the cisplatin 10 mg·kg^(-1)·d^(-1) group were significantly decreased compared with those in the control group(P<0.05,P<0.01);and the vertical movement of the cisplatin 3 and 5 mg·kg^(-1)·d^(-1) groups was also decreased compared with that of the control group(P<0.01).(2)Com

关 键 词：顺铂 虚证 药源性证候 肾上腺皮质 睾丸 小鼠 

分 类 号：R285.5[医药卫生—中药学]