1. 上海中医药大学基础医学院,上海,201203
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潘志强, 梁龙龙, 王晓敏, 等. 氢化可的松诱发虚证模型小鼠的证候特征再评价及肾上腺皮质功能变化研究[J]. 上海中医药杂志, 2017,51(8):13-20.
PAN Zhi-qiang, LIANG Long-long, WANG Xiao-min, et al. Study on the characteristics of mice model with deficiency syndrome induced by hydrocortisone and changes of adrenocortical function[J]. Shanghai Journal of Traditional Chinese Medicine, 2017,51(8):13-20.
潘志强, 梁龙龙, 王晓敏, 等. 氢化可的松诱发虚证模型小鼠的证候特征再评价及肾上腺皮质功能变化研究[J]. 上海中医药杂志, 2017,51(8):13-20. DOI:
PAN Zhi-qiang, LIANG Long-long, WANG Xiao-min, et al. Study on the characteristics of mice model with deficiency syndrome induced by hydrocortisone and changes of adrenocortical function[J]. Shanghai Journal of Traditional Chinese Medicine, 2017,51(8):13-20. DOI:
目的:研究氢化可的松诱发虚证模型小鼠的证候特征及其可能的物质基础。 方法:将60只雄性昆明种小鼠随机分为正常对照组和模型组,每组30只。正常对照组自由饮水与摄食,模型组自由饮用剂量递减的氢化可的松溶液。其中,0.015 g·kg-1·d-1剂量,饮用3天;0.010 g·kg-1·d-1剂量,饮用5天;0.007 5 g·kg-1·d-1剂量,饮用7天。在造模的第8天、第15天,采用本课题组建立的小鼠四诊标准采集技术与辨证方法判断小鼠的证候属性;于造模的第3天、第8天、第15天分批处死小鼠,取肾上腺组织,分别抽提RNA和蛋白质,以实时荧光定量PCR技术检测相关基因表达,以Western blotting法检测类固醇合成急性调节蛋白(STAR)表达,并以ELISA方法检测血清促肾上腺皮质激素(ACTH)含量。 结果:①与正常对照组比较,模型组小鼠在造模期间,消瘦、体质量增长显著减慢(P<0.05),被毛蓬松易脱落、光泽度下降,活动度降低、喜蜷缩,体温尤其是尾部温度下降,爪部色泽变化不明显。②造模第15天,与正常对照组比较,模型组小鼠气虚证、阴虚证明显,肾上腺、脾脏与胸腺持续性萎缩(P<0.05);血液ACTH水平下降(P<0.05);肾上腺皮质激素合成过程重要受体Srb1、Ldlr基因表达降低(P<0.01),类固醇激素合成酶Star、Cyp11a1、Cyp21a1、Cyp11b1、Cyp11b2基因表达及其酶的调节因子Fdx1、Fdxr、Nr4a1、Nr4a2、Nr5a1基因表达显著下降(P<0.05),以及STAR蛋白表达水平明显降低(P<0.01)。 结论:氢化可的松复制的小鼠模型属于虚证模型,具有气虚与阴虚证的特征;参与肾上腺皮质激素合成的重要功能分子涉及相关证候的发生。
Objective:To study the characteristics of mice model with deficiency syndrome induced by hydrocortisone and the potential material base. Methods 60 male Kunming mice were randomly divided into the normal control group and model group,30 mice in each group. The mice in the normal control group were treated with normal water and feeding,and the mice in the model group were treated with hydrocortisone solution with degressive dose(0.015 g·kg-1·d-1 for 3 days,0.010 g·kg-1·d-1 for 5 days and 0.007 5 g·kg-1·d-1 for 7 days). On the eighth and fifteenth day,the syndrome classification of mice was determined by standard acquisition technique and syndrome differentiation methods for the four methods of diagnosis in mice that created by our research group. On the third,eighth and fifteenth day,the mice were sacrificed respectively. The adrenal gland was collected and the RNA and protein were extracted. The expressions of related genes were detected by real time fluorescent quantitative PCR,the expression of steroidogenic acute regulatory protein(STAR) was detected by Western blotting,and the content of serum adrenocorticotropic hormone (ACTH) was measured by ELISA. Results: ①Compared with the normal control group,the body weight of the mice in the model group was significantly decreased during the modeling period(P<0.05),and the appearance such as marasmus,fluffy hair falling,decreased glossiness of hair,reduced activity with curled posture were observed,the temperature especially in the tail was decreased,but there was no obvious change on the claw color. ②Compared with the normal control group on the fifteenth day,the mice in the model group showed obvious syndrome of qi deficiency and yin deficiency,the persistent atrophy of adrenal gland,spleen and thymus were observed(P<0.05). The serum level of ATCH was decreased in the model group(P<0.05),the gene expressions of Srb1 and Ldlr as the important receptor in adrenocortical hormone synthesis were decreased(P<0.01),the gene expressions of steroid hormone synthases such as Star,Cyp11a1,Cyp21a1,Cyp11b1,Cyp11b2 and their transcription factors such as Fdx1,Fdxr,Nr4a1,Nr4a2 and Nr5a1 were significantly decreased (P<0.05), and the protein expression level of StAR was significantly decreased(P<0.01). Conclusion: Hydrocortisone can be used to induce the mice model with deficiency syndrome,which shows the characteristics of syndrome of qi deficiency and yin deficiency. The important functional molecules in the adrenocortical hormone synthesis are associated with the related syndrome.
证候虚证疾病模型小鼠氢化可的松肾上腺皮质功能
syndromedeficiency syndromedisease modelmicehydrocortisoneadrenocortical function
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