顺铂连续和间隔造模诱发的药源性虚证小鼠类固醇激素合成能力的比较研究  

Comparative study of steroid hormone synthesis in a drug-induced deficiency syndrome by continuous and interval administration of cisplatin in mice

作　　者：农淄心 邓琳[2] 张超峰 贺健祥 谢海纳 潘志强[1] NONG Zixin;DENG Lin;ZHANG Chaofeng;HE Jianxiang;XIE Haina;PAN Zhiqiang(School of Basic Medical Sciences,Shanghai University of Traditional Chinese Medicine,Shanghai 201203;China.2.Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203)

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：《中国实验动物学报》2021年第5期626-636,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：上海市科委实验动物专项(19140905000)。

摘　　要：目的研究顺铂不同给药方案对小鼠类固醇激素合成及储备功能的影响。方法40只雌性ICR小鼠分为4组,顺铂连续和间隔造模组以及两个对照组。顺铂连续造模组给药7 d,每天注射3 mg/kg顺铂;间隔造模组每周注射1次10 mg/kg顺铂,共4次。末次造模前进行旷场实验、转棒疲劳实验、抓力测试;次日处死前称体重,摘取小鼠心脏、脾、胸腺和双侧肾并称重;RT-qPCR检测下丘脑GnRH和CRH、垂体Pomc、Fshb、Lhb、Pou1f1基因表达,以及肾上腺和卵巢类固醇合成酶(St AR、Cyp11a1、Cyp21a1、Cyp17a1、Cyp11b1、Hsd3b2、Cyp19a1、Hsd17b1)及受体(Esr1、Esr2和Gper1)基因表达;Western Blot检测肾上腺和卵巢St AR、CYP11A1、CYP21A2和CYP11B1蛋白表达。结果与对照组比较:(1)不同频次顺铂造模均可抑制小鼠体重增长,顺铂累积后可以诱导小鼠气虚证表现,并抑制脏器质量。(2)不同频次顺铂造模均能加速激活原始卵泡分化,产生大量闭锁卵泡,诱导膜细胞及颗粒细胞凋亡,健康的成熟卵泡减少;但对肾上腺组织和细胞形态影响不大。(3)连续造模后肾上腺St AR蛋白表达下调;间隔造模后St AR升高,但下调CYP11B1蛋白表达。(4)连续造模后,卵巢Cyp11a1、Hsd3b2、Esr1和Gper1基因表达上调,Star和Cyp17a1基因表达下调;间隔造模后St AR蛋白表达和Hsd3b2基因表达下调。(5)顺铂连续造模后下丘脑GnRH和CRH基因表达明显下调。(6)顺铂连续造模后垂体Pomc、Fshb、Pou1f1基因表达升高,而间隔造模组Lhb表达下降。结论顺铂能够诱发小鼠气虚、精气不足证的相关表现;合成类固醇激素的卵巢受到选择性损伤大于肾上腺,短期连续给药也进一步抑制HPA轴和HPG轴的高级中枢下丘脑和垂体功能。Objective To research the effect of different administration protocols of cisplatin on the biosynthesis and reserve of steroids in mice.Methods Forty female ICR mice were randomly divided into four groups:a cisplatin continuous administration group(CA-group),a cisplatin interval administration group(IA-group),and two corresponding control groups.The CA-group was injected with 3 mg/kg cisplatin for 7 consecutive days.The IA-group was injected with 10 mg/kg cisplatin once per week,for 4 weeks.Rotating-stick,holding power,movements and axillary temperature were examined before the last injection,also the end of laboratory animal model making.The day after,the mice were killed,the visceral organs were separated weighed,included the kidney,heart,thymus and spleen.Relative mRNA expression levels in the hypothalamus(GnRH and CRH),pituitary(Pomc,Fshb,Lhb,and Pou1 f1),adrenal and ovary(St AR,Cyp11 a1,Cyp21 a1,Cyp17 a1,Cyp11 b1,Hsd3 b2,Cyp19 a1,Hsd17 b1,Esr1,Esr2,and Gper1)tissue were examined by RT-qPCR.Protein expression levels(St AR,CYP11 A1,CYP21 A2 and CYP11 B1)in adrenal and ovary tissues were examined.Results Compared with the control group,body weight was reduced by cisplatin in two different mouse models.Visceral organs were inhibited and the Qi deficiency syndrome was induced by cisplatin administration.In two different cisplatin models,murine primordial follicles showed accelerated activation and differentiation,and many atretic follicles were produced containing apoptotic granulosa and theca cells.Healthy mature follicle numbers were reduced significantly.There were no detectable changes in adrenal histology.In adrenal tissue,St AR expression was inhibited in the CA-group and upregulated in the IA-group,which also exhibited reduced CYP11 B1 protein expression.In the ovary,the expression levels of Cyp11 a1,Hsd3 b2,Esr1,and Gper1 mRNAs were upregulated and Star and Cyp17 a1 mRNA expression was reduced in the CA-group.The expression levels of St AR protein and Hsd3 b2 mRNA were decreased in the IA-group.In the hyp

关 键 词：顺铂 证候 药源性证候 卵巢 肾上腺 类固醇激素 

分 类 号：Q95-33[生物学—动物学]