MiRNA-206参与甲状腺功能亢进症患者代谢异常的机制研究
发布时间：2018-10-05 06:58
【摘要】：目的:探讨甲状腺功能亢进症(甲亢)患者体内循环微小核糖核酸-206(micro RNA-206,mi R-206)的表达变化及其参与甲状腺激素(thyroid hormone,TH)调节脂代谢的相关机制。方法:按照纳入和排除标准收集2013年10月~2014年03月来自我院20-60岁15例内分泌科门诊的甲亢患者和12例体检中心的健康者血清标本,采用实时定量聚合酶链式反应(quantitative real-time Polymerase Chain Reaction,q RT-PCR)技术检测血清标本中mi R-206表达水平的变化并利用线性回归分析其与游离三碘甲状腺原氨酸(free triiodothyronine,FT3)、游离四碘甲状腺原氨酸(free tetraiodothyronine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)相关性;另外以肝癌细胞株(Hep G2细胞)为工具,在其培养基内添加三碘甲状腺原氨酸(triiodothyronine,T3)干预,采用q RT-PCR技术检测细胞内mi R-206表达水平变化,并分别在生化分析仪上采用酶偶联比色法检测细胞内甘油三酯(triglycerides,TG)含量变化和利用油红进行染色;在细胞过表达/沉默mi R-206后,采用酶偶联比色法检检测细胞内TG含量的变化;在T3干预过表达mi R-206的细胞后,采用酶偶联比色法检检测细胞内TG含量的变化,观察T3对过表达mi R-206细胞内TG含量的变化。结果:1.q RT-PCR技术检测结果表明:甲亢患者血清内mi R-206的表达水平明显低于健康者(p0.05)且血清内mi R-206水平与FT3浓度呈负相关(r=㧟0.408,p=0.035),与FT4浓度呈负相关(r=㧟0.451,p=0.018),与TSH浓度呈正相关(r=0.445,p=0.020);2.采用T3分别干预Hep G2细胞24h、36h后,细胞内mi R-206表达水平均明显降低(p0.05);3.采用T3分别干预Hep G2细胞12h、24h、36h后,TG含量均明显降低(p0.01);4.沉默/过表达mi R-206后,细胞内TG含量显著性降低/升高(p0.05);5.用T3干预过表达mi R-206和阴性对照(negative control,NC)的Hep G2细胞,过表达mi R-206细胞内TG含量显著性高于NC,说明过表达mi R-206具有拮抗T3降低TG生成的作用(p0.05)。结论:1.甲亢患者血清mi R-206含量显著降低;2.人血清mi R-206含量与TH浓度呈负相关关系;3.Mi R-206对Hep G2细胞TG代谢具有调控作用;4.Mi R-206可能参与甲亢患者体内T3对TG代谢的调节过程。
[Abstract]:Aim: to investigate the changes of circulating microribonucleic acid (micro RNA-206,mi R-206) expression in patients with hyperthyroidism (hyperthyroidism) and the mechanisms involved in the regulation of lipid metabolism by thyroid hormone (thyroid hormone,TH). Methods: from October 2013 to March 2014, 15 patients with hyperthyroidism from 20 to 60 years old and 12 healthy people from physical examination center were collected according to the inclusion and exclusion criteria. Real time quantitative polymerase chain reaction (quantitative real-time Polymerase Chain Reaction,q RT-PCR) was used to detect the expression of mi R-206 in serum samples and its relationship with free triiodothyronine (free triiodothyronine,FT3) and free tetraiodothyronine was analyzed by linear regression analysis. (free tetraiodothyronine,FT4), thyroid stimulating hormone (thyroid stimulating hormone,TSH) correlation; In addition, the hepatoma cell line (Hep G2 cell) was used as a tool and triiodothyronine (triiodothyronine,T3) was added to the medium. The expression of mi R-206 was detected by Q RT-PCR technique. The content of triglyceride (triglycerides,TG) in cells was detected by enzyme-coupled colorimetry and stained with oil red on biochemical analyzer, and the changes of TG content in cells were detected by enzyme-coupled colorimetry after over-expression / silencing of mi R-206. After T3 intervention, the changes of TG content in cells expressing mi R-206 were detected by enzyme-coupled colorimetric assay, and the changes of TG content in over-expressed mi R-206 cells were observed. Results 1. The expression of mi R-206 in serum of hyperthyroidism patients was significantly lower than that of healthy controls (p0.05), and there was a negative correlation between the mi R-206 level and FT3 concentration in hyperthyroidism patients. The concentration of FT4 was negatively correlated with the concentration of FT4. There was a positive correlation between the concentration of TSH and the concentration of TSH (r = 0.445P0. 020). After treated with T3 for 24 h or 36 h, the expression of mi R-206 in Hep G2 cells decreased significantly (p0.05). Triglyceride content in Hep G2 cells decreased significantly (p0.01) after treatment with T3 for 12 h, 24 h and 36 h, respectively. After silencing / overexpression of mi R-206, the content of TG in the cells decreased / increased significantly (p0.05). The overexpression of TG in mi R-206 cells was significantly higher than that in NC, cells, which indicated that the overexpression of mi R-206 could antagonize T3 and TG production in Hep G2 cells (p0.05). Conclusion 1. Serum mi R-206 decreased significantly in patients with hyperthyroidism. There was a negative correlation between serum mi R-206 and TH concentration. Mi R-206 could regulate TG metabolism in Hep G2 cells. 4. Mi R-206 may be involved in the regulation of TG metabolism by T3 in patients with hyperthyroidism.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R581.1
本文编号：2252445
[Abstract]:Aim: to investigate the changes of circulating microribonucleic acid (micro RNA-206,mi R-206) expression in patients with hyperthyroidism (hyperthyroidism) and the mechanisms involved in the regulation of lipid metabolism by thyroid hormone (thyroid hormone,TH). Methods: from October 2013 to March 2014, 15 patients with hyperthyroidism from 20 to 60 years old and 12 healthy people from physical examination center were collected according to the inclusion and exclusion criteria. Real time quantitative polymerase chain reaction (quantitative real-time Polymerase Chain Reaction,q RT-PCR) was used to detect the expression of mi R-206 in serum samples and its relationship with free triiodothyronine (free triiodothyronine,FT3) and free tetraiodothyronine was analyzed by linear regression analysis. (free tetraiodothyronine,FT4), thyroid stimulating hormone (thyroid stimulating hormone,TSH) correlation; In addition, the hepatoma cell line (Hep G2 cell) was used as a tool and triiodothyronine (triiodothyronine,T3) was added to the medium. The expression of mi R-206 was detected by Q RT-PCR technique. The content of triglyceride (triglycerides,TG) in cells was detected by enzyme-coupled colorimetry and stained with oil red on biochemical analyzer, and the changes of TG content in cells were detected by enzyme-coupled colorimetry after over-expression / silencing of mi R-206. After T3 intervention, the changes of TG content in cells expressing mi R-206 were detected by enzyme-coupled colorimetric assay, and the changes of TG content in over-expressed mi R-206 cells were observed. Results 1. The expression of mi R-206 in serum of hyperthyroidism patients was significantly lower than that of healthy controls (p0.05), and there was a negative correlation between the mi R-206 level and FT3 concentration in hyperthyroidism patients. The concentration of FT4 was negatively correlated with the concentration of FT4. There was a positive correlation between the concentration of TSH and the concentration of TSH (r = 0.445P0. 020). After treated with T3 for 24 h or 36 h, the expression of mi R-206 in Hep G2 cells decreased significantly (p0.05). Triglyceride content in Hep G2 cells decreased significantly (p0.01) after treatment with T3 for 12 h, 24 h and 36 h, respectively. After silencing / overexpression of mi R-206, the content of TG in the cells decreased / increased significantly (p0.05). The overexpression of TG in mi R-206 cells was significantly higher than that in NC, cells, which indicated that the overexpression of mi R-206 could antagonize T3 and TG production in Hep G2 cells (p0.05). Conclusion 1. Serum mi R-206 decreased significantly in patients with hyperthyroidism. There was a negative correlation between serum mi R-206 and TH concentration. Mi R-206 could regulate TG metabolism in Hep G2 cells. 4. Mi R-206 may be involved in the regulation of TG metabolism by T3 in patients with hyperthyroidism.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R581.1
【参考文献】
相关期刊论文 前1条
1 王穗;张春风;;儿童甲状腺功能亢进症52例[J];实用儿科临床杂志;2007年14期
，本文编号：2252445
本文链接：https://www.wllwen.com/yixuelunwen/nfm/2252445.html
最近更新
教材专著
