【摘要】：目的:探討甲狀腺功能亢進癥(甲亢)患者體內(nèi)循環(huán)微小核糖核酸-206(micro RNA-206,mi R-206)的表達變化及其參與甲狀腺激素(thyroid hormone,TH)調(diào)節(jié)脂代謝的相關(guān)機制。方法:按照納入和排除標(biāo)準(zhǔn)收集2013年10月~2014年03月來自我院20-60歲15例內(nèi)分泌科門診的甲亢患者和12例體檢中心的健康者血清標(biāo)本,采用實時定量聚合酶鏈?zhǔn)椒磻?yīng)(quantitative real-time Polymerase Chain Reaction,q RT-PCR)技術(shù)檢測血清標(biāo)本中mi R-206表達水平的變化并利用線性回歸分析其與游離三碘甲狀腺原氨酸(free triiodothyronine,FT3)、游離四碘甲狀腺原氨酸(free tetraiodothyronine,FT4)、促甲狀腺激素(thyroid stimulating hormone,TSH)相關(guān)性;另外以肝癌細(xì)胞株(Hep G2細(xì)胞)為工具,在其培養(yǎng)基內(nèi)添加三碘甲狀腺原氨酸(triiodothyronine,T3)干預(yù),采用q RT-PCR技術(shù)檢測細(xì)胞內(nèi)mi R-206表達水平變化,并分別在生化分析儀上采用酶偶聯(lián)比色法檢測細(xì)胞內(nèi)甘油三酯(triglycerides,TG)含量變化和利用油紅進行染色;在細(xì)胞過表達/沉默mi R-206后,采用酶偶聯(lián)比色法檢檢測細(xì)胞內(nèi)TG含量的變化;在T3干預(yù)過表達mi R-206的細(xì)胞后,采用酶偶聯(lián)比色法檢檢測細(xì)胞內(nèi)TG含量的變化,觀察T3對過表達mi R-206細(xì)胞內(nèi)TG含量的變化。結(jié)果:1.q RT-PCR技術(shù)檢測結(jié)果表明:甲亢患者血清內(nèi)mi R-206的表達水平明顯低于健康者(p0.05)且血清內(nèi)mi R-206水平與FT3濃度呈負(fù)相關(guān)(r=㧟0.408,p=0.035),與FT4濃度呈負(fù)相關(guān)(r=㧟0.451,p=0.018),與TSH濃度呈正相關(guān)(r=0.445,p=0.020);2.采用T3分別干預(yù)Hep G2細(xì)胞24h、36h后,細(xì)胞內(nèi)mi R-206表達水平均明顯降低(p0.05);3.采用T3分別干預(yù)Hep G2細(xì)胞12h、24h、36h后,TG含量均明顯降低(p0.01);4.沉默/過表達mi R-206后,細(xì)胞內(nèi)TG含量顯著性降低/升高(p0.05);5.用T3干預(yù)過表達mi R-206和陰性對照(negative control,NC)的Hep G2細(xì)胞,過表達mi R-206細(xì)胞內(nèi)TG含量顯著性高于NC,說明過表達mi R-206具有拮抗T3降低TG生成的作用(p0.05)。結(jié)論:1.甲亢患者血清mi R-206含量顯著降低;2.人血清mi R-206含量與TH濃度呈負(fù)相關(guān)關(guān)系;3.Mi R-206對Hep G2細(xì)胞TG代謝具有調(diào)控作用;4.Mi R-206可能參與甲亢患者體內(nèi)T3對TG代謝的調(diào)節(jié)過程。
[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.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R581.1

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1 王穗;張春風(fēng);;兒童甲狀腺功能亢進癥52例[J];實用兒科臨床雜志;2007年14期

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