淫羊藿苷改善甲状腺功能减退模型大鼠的垂体发育  

Icariin improves pituitary development in hypothyroidism model rats

作　　者：白改改 赵玉娟[2] 魏家凯 黄文娣 安瑶 王志 Bai Gaigai;Zhao Yujuan;Wei Jiakai;Huang Wendi;An Yao;Wang Zhi(Department of Endocrine Genetics and Metabolism,Xi’an Children’s Hospital,Xi’an,Shaanxi 710000,China;Department of Neonatology,Xi’an Children’s Hospital,Xi’an,Shaanxi 710000,China)

机构地区：[1]西安市儿童医院内分泌遗传代谢科,陕西省西安市710000 [2]西安市儿童医院新生儿科,陕西省西安市710000

出　　处：《中国组织工程研究》2023年第28期4546-4553,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:甲状腺功能减退常伴随着垂体合成分泌激素分泌不足,有研究表明淫羊藿苷对甲状腺功能减退引起的垂体激素分泌不足有确切疗效,但其作用机制还不清楚。目的:探究淫羊藿苷对大鼠甲状腺功能减退后垂体合成分泌激素的影响及机制。方法:(1)40只新生雌性SD大鼠随机分为对照组、模型组及75,150,300 mg/kg淫羊藿苷组,每组8只,后4组给予丙基硫氧嘧啶1.5 mg/(kg·d)灌胃处理构建甲状腺功能减退大鼠模型,各淫羊藿苷剂量组给予75,150,300 mg/kg淫羊藿苷干预治疗,检测各组miR-17-5p、LIM同源框4(LIM homeobox 4,LHX4)、PROP成对同源1蛋白(prop paired end homology 1 protein,PROP1)和HESX1表达,以及垂体合成分泌激素含量。(2)采用锂盐处理共培养的大鼠甲状腺细胞和垂体前叶细胞,给予不同浓度淫羊藿苷(10,30,50,70,100μmol/L)干预,CCK-8试剂盒检测垂体前叶细胞活力;ELISA试剂盒检测促卵泡激素、生长激素和促甲状腺激素水平;实时荧光定量PCR检测miR-17-5p表达;Western印迹法检测PROP1和HESX1蛋白水平;采用双荧光素报告酶基因和RNA结合蛋白免疫沉淀实验检测miR-17-5p和LHX4的结合关系。结果与结论:(1)体内实验结果显示,150 mg/kg淫羊藿苷组较模型组促甲状腺激素含量降低(P<0.05),生长激素(P<0.01)和促卵泡激素(P<0.05)含量显著升高;此外,150mg/kg淫羊藿苷组较模型组血清游离三碘甲状腺原氨酸、血清游离甲状腺素、血清三碘甲状腺原氨酸和血清甲状腺素激素分泌水平增加;(2)用50,70,100μmol/L淫羊藿苷干预能够增强大鼠垂体前叶细胞活力(P<0.01),其中70μmol/L淫羊藿苷干预效果最佳(P均<0.01);(3)淫羊藿苷处理抑制了锂盐处理的大鼠垂体前叶细胞中miR-17-5p表达增加,上调PROP1和HESX1表达(P均<0.05);(4)锂盐处理组细胞上清液中生长激素和促卵泡激素含量明显降低,促甲状腺激素含量增加,与对照组相比差异有显著性意义(P<0.01)BACKGROUND:Hypothyroidism is often accompanied by inadequate secretion of pituitary synthetic secretory hormones.Some studies have shown that icariin has an exact effect on pituitary hormone secretion deficiency caused by hypothyroidism,but the mechanism is still unclear.OBJECTIVE:To investigate the effect and mechanism of icariin on pituitary hormone synthesis and secretion after hypothyroidism in rats.METHODS:(1)Forty newborn female Sprague-Dawley rats were randomized into control,model,75,150,300 mg/kg icariin groups with 8 rats in each group.The rats in the latter four groups were intragastrically treated with propylthiouracil(1.5 mg/kg/day)to construct a hypothyroid rat model,and treated with different doses(0,75,150,and 300 mg/kg)of icariin.The expression of miR-17-5p,LIM homeobox 4(LHX4),prop paired end homology 1 protein(PROP1),and HESX1 was detected,as well as hormone content synthetized and secreted by the pituitary gland.(2)Co-cultured rat thyrocytes and anterior pituitary cells were treated with lithium salt,and different concentrations of icariin(10,30,50,70,100μmol/L)were administe red.Anterior pituitary cell viability was assessed by cell counting kit-8.Follicle-stimulating hormone,growth hormone,and thyroid-stimulating hormone levels were detected by ELISA kits.miR-17-5p expression was detected by real-time fluorescence quantitative PCR.PROP1 and HESX1 protein levels were measured by western blot assay.The binding relationship between miR-17-5p and LHX4 was examined by RNA binding protein immunoprecipitation and dual luciferin reporter assays.RESULTS AND CONCLUSION:In vivo results showed that the treatment with 150 mg/kg icariin decreased the contents of thyroid-stimulating hormone and significantly increased the contents of growth hormone and follicle-stimulating hormone compared with the model group.In addition,the hormone secretion levels of serum free triiodothyronine,serum free thyroxine,serum triiodothyronine,and serum thyroxine were increased in the 150 mg/kg icariin group compared with th

关 键 词：淫羊藿苷 甲状腺功能减退 miR-17-5p/LHX4轴 生长素 促甲状腺激素 促卵泡激素 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R722.1