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不同剂量氢化可的松诱发小鼠药源性脾肾虚证的评价研究

Study on Drug-induced Pi and Shen Deficiency Syndrome of Mice Induced by Different Doses of Hydrocortisone
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摘要 目的评价氢化可的松诱发小鼠药源性脾肾虚证模型,并探索其物质基础。方法 80只ICR小鼠按随机分为对照组、极低、低、中、高剂量组,每组16只,分别采用灭菌水、0.33、0.66、3.3、12.5 mg/(kg·d)氢化可的松灌胃。每日动态观测小鼠体重,并分别在给药第7天和第14天,检测小鼠抓力、旷场活动度、躯体不同部位红外温度等表征信息。分别于给药第8天和第15天各处死小鼠8只,取脾脏、胸腺、心脏和肾脏称重并计算脏器指数;ELISA检测血清皮质酮(CORT)和促肾上腺皮质激素(ACTH)含量;实时荧光定量PCR检测肾上腺StAR、Cyp11a1、Cyp21a1、Cyp11b1、Cyp11b2 mRNA表达;Western Blot检测肾上腺LDLR、SRBI、StAR蛋白表达。结果与对照组同期比较,低、中剂量组4~14天体重下降(P<0.05),极低剂量组7天尾部最低温度、StAR、Cyp21a1 mRNA表达,14天头部最高温度、躯干平均温度、尾部最低温度以及ACTH含量、LDLR、StAR蛋白表达下降(P<0.05,P<0.01),14天抓力上升(P<0.05);低、中、高剂量组7、14天头部最高温度、躯干平均温度、尾部最低温度、胸腺重量和指数下降(P<0.05,P<0.01);低剂量组7、14天StAR mRNA、LDLR、StAR表达下降(P<0.05,P<0.01),14天CORT含量、ACTH、Cyp11a1、Cyp11b1 mRNA表达下降(P<0.05,P<0.01);中、高剂量组7、14天脾脏重量和指数、CORT含量、ACTH、StAR、Cyp21a1、Cyp11b1 mRNA、LDLR、SRBI、StAR表达下降(P<0.05,P<0.01),14天Cyp11a1、Cyp11b2 mRNA下降(P<0.05,P<0.01);高剂量组14天体重、旷场活动度下降(P<0.05)。结论采用0.66及3.3 mg/(kg·d)氢化可的松持续给药14天可建立稳定的药源性脾肾虚证小鼠模型,可能与其降低肾上腺皮质激素合成酶表达和垂体-肾上腺皮质轴功能有关。 Objective To evaluate Pi and Shen deficiency syndrome of mice induced by different doses of hydrocortisone,and explore the material basis.Methods Totally 80 ICR mice were randomly divided into the nomal group,extremly low dose group,low dose group,medium dose group,high dose group,16 in each group,they were respectively administrated with sterile water,0.33,0.66,3.3 and 12.5 mg/kg-1·d-1 hydrocortisone by gastrogavage.The weight of mice was dynamically observed every day,and the characteristic information such as holding power,activity of open field and infrared temperature of different parts of the body were detected on at the 7th and 14th day.8 mice were sacrificed on 8th and 15th day repectively,whose spleen,thymus,heart and kidney were weighed and the organ index was calculated.Serum corticosterone(CORT)and adrenocorticotrophic hormone(ACTH)were measured by ELISA.The mRNA expressions of StAR,Cyp11a1,Cyp21a1,Cyp11b1 and Cyp11b2 in the adrenal were detected by real-time fluorescent quantitative PCR.The proteins expressions of LDLR,SRBI and StAR were detected by Western Blot.Results Compared with the control group in the same time,the weight decreased in low and medium dose groups from 4th to 14th day(P<0.05).Tail minimum temperature,StAR,Cyp21a1 mRNA expression at 7th day decreased and head maximum temperature,trunk mean temperature,tail minimum temperature,the levels of ACTH,LDLR,StAR at 14th day decreased in extremely low dose group(P<0.05,P<0.01),while holding power at 14th day increased(P<0.05);head maximum temperature,trunk mean temperature,tail minimum temperature,thymus weight and index at 7th and 14th day decreased in low,medium and high dose groups(P<0.05,P<0.01);the expressions of StAR,LDLR,StAR at 7th and 14th day and corticosterone,ACTH,Cyp11a1 and Cyp11b1 mRNA expression at 14th day decreased in the low dose group(P<0.05,P<0.01),and corticosterone,ACTH,Cyp11a1 and Cyp11b1 at 14 days decreased(P<0.05,P<0.01);spleen weight and index,corticosterone,ACTH,StAR,Cyp21a1,Cyp11b1 mRNA and LDLR,SRBI,StAR at
作者 潘志强 钱宏梁 王晓敏 卢涛 李亚 方肇勤 PAN Zhi-qiang;QIAN Hong-liang;WANG Xiao-min;LU Tao;LI Ya;FANG Zhao-qin(Basic Medical School of Shanghai University of Traditional Chinese Medicine,Shanghai 201203)
出处 《中国中西医结合杂志》 CAS CSCD 北大核心 2019年第4期480-485,共6页 Chinese Journal of Integrated Traditional and Western Medicine
基金 国家自然科学基金面上项目(No.81473562,No.81873212).
关键词 氢化可的松 肾上腺皮质 证候 药源性证候 虚证 动物模型 hydrocortisone adrenal cortex syndromes drug-induced syndromes deficiency syndromes animal model
作者简介 通讯作者:潘志强,Tel:021-51322116,E-mail:pzq527@163.com
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