本文選題：T細(xì)胞發(fā)育 + 亞臨床甲狀腺功能減退��； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：研究背景:促甲狀腺素(thyrotropin,TSH)被認(rèn)為僅僅甲狀腺調(diào)節(jié)。然而,新的證據(jù)表明,高血清TSH水平可能存在潛在的有害的影響。高血清TSH水平,是非酒精性脂肪肝病和心血管疾病的獨(dú)立危險(xiǎn)因素,而且往往伴隨著慢性炎癥、胰島素抵抗、氧化應(yīng)激。亞臨床甲狀腺功能減退癥(subclinical hypothyroidism,SCH),是一種臨床上較為常見的隱匿性疾病,其特點(diǎn)是甲狀腺功能血清學(xué)檢測(cè)表現(xiàn)現(xiàn)為促甲狀腺激素水平增高,但游離甲狀腺素水平尚正常,是甲狀腺機(jī)能減退障礙的早期階段。亞甲減癥的發(fā)病率呈現(xiàn)逐漸升高的趨勢(shì),并對(duì)健康有潛在的害影響。SCH在成年人中的患病率約為4-100%。胸腺是重要的中樞免疫器官,是T淋巴細(xì)胞離開骨髓后發(fā)育的重要場(chǎng)所。胸腺為T淋巴細(xì)胞的發(fā)育,提供了一個(gè)特定的微環(huán)境,既胸腺微環(huán)境。對(duì)胸腺內(nèi)T細(xì)胞發(fā)育、分化以及成熟的研究,是當(dāng)今免疫學(xué)研究的重要熱門領(lǐng)域。研究表明,T細(xì)胞表達(dá)促甲狀腺激素受體(TSHreceptor,TSH-R),而SCH中胸腺T細(xì)胞發(fā)育的變化尚未完全闡明。研究目的:本實(shí)驗(yàn)旨在闡明亞甲減小鼠模型中,TSH對(duì)胸腺T細(xì)胞發(fā)育的影響,SCH小鼠模型中胸腺的細(xì)胞組成情況以及不同發(fā)育階段的T細(xì)胞凋亡變化。研究方法:1.SCH小鼠模型的構(gòu)建:應(yīng)用甲巰基咪唑喂養(yǎng)小鼠,甲巰基咪唑劑量為0.08mg/kg/d,甲巰基咪唑喂養(yǎng)16周時(shí),采用I125標(biāo)記放射免疫的方法,檢測(cè)小鼠的血清中游離甲狀腺素(FT4)水平,采取Elisa的方法測(cè)驗(yàn)小鼠血清TSH水平。通過評(píng)估小鼠的血清甲狀腺功能,來構(gòu)建SCH小鼠模型,使其血清檢測(cè)符合SCH診斷標(biāo)準(zhǔn),既血清TSH水平升高,甲狀腺激素水平無顯著變化的,研究TSH對(duì)T細(xì)胞發(fā)育的影響。2.SCH小鼠胸腺中不同發(fā)育階段T的免疫學(xué)組成分析:應(yīng)用多色彩流式分析的方法檢測(cè)小鼠胸腺中T細(xì)胞不同發(fā)育階段各T細(xì)胞亞群的比例。3.胸腺中不同發(fā)育階段T細(xì)胞凋亡比例的測(cè)定:應(yīng)用凋亡試劑盒檢測(cè)各T細(xì)胞亞群細(xì)胞凋亡的比例。4.小鼠胸腺內(nèi)凋亡相關(guān)蛋白檢測(cè):采取Western Blot法測(cè)驗(yàn)胸腺p-ERK/ERK的蛋白水平變化。5.胸腺近期輸出功能檢測(cè):提取小鼠脾臟DNA,實(shí)時(shí)定量PCR檢測(cè)小鼠脾臟T細(xì)胞DNA中TREC水平。結(jié)果:1.SCH小鼠模型的構(gòu)建:甲巰基咪唑喂養(yǎng)小鼠16周,SCH組小鼠血清FT4測(cè)值與對(duì)照組小鼠相比,無明顯差異性的變化(P0.05);SCH模型組血清TSH水平明顯高于對(duì)照組小鼠(P0.05),兩組血清TSH水平差異具有統(tǒng)計(jì)學(xué)意義。2.SCH小鼠胸腺中不同發(fā)育階段T的免疫學(xué)組成分析:SCH小鼠胸腺重量比對(duì)照組增加18%。胸腺中CD4 +和CD8 +單陽性(single positive,SP)胸腺細(xì)胞分別增加了38%和44%。3.胸腺中不同發(fā)育階段T細(xì)胞凋亡比例的測(cè)定:通過檢測(cè)凋亡,我們發(fā)現(xiàn)在SCH小鼠胸腺中,Annexin-V+細(xì)胞比例減少(p0.05),證明TSH減少SCH小鼠胸腺T細(xì)胞凋亡。4.小鼠胸腺內(nèi)凋亡相關(guān)蛋白檢測(cè):Western blot結(jié)果顯示,SCH小鼠胸腺中p-ERK 1/2增加,表明SCH小鼠胸腺組織中ERK通路被激活。5.胸腺近期輸出功能檢測(cè):PCR檢測(cè)小鼠脾臟組織T細(xì)胞受體切除環(huán)(TREC),結(jié)果顯示,SCH組小鼠脾臟T細(xì)胞中TREC增加(p0.05)。結(jié)論及意義:這些結(jié)果表明,TSH促進(jìn)SCH小鼠胸腺T細(xì)胞發(fā)育,增強(qiáng)胸腺近期輸出。其機(jī)制可能是通過抑制胸腺細(xì)胞的凋亡,抗凋亡作用可能是通過p-ERK/ERK信號(hào)通路。
[Abstract]:Background: thyrotropin (TSH) is thought to be only thyroid regulation. However, new evidence suggests that high serum TSH levels may have potentially harmful effects. High serum TSH levels are independent risk factors for nonalcoholic fatty liver disease and cardiovascular disease, and are often accompanied by chronic inflammation, insulin resistance, and oxidation. Stress. Subclinical hypothyroidism (subclinical hypothyroidism, SCH) is a clinically common occult disease characterized by thyroid function serological detection showing elevated levels of thyroid stimulating hormone, but free thyroxine level Shang Zhengchang is an early stage of hypothyroidism. Subhypothyroidism is a subgrade of hypothyroidism. The incidence of the disease is increasing gradually, and the potential harm to health of the.SCH in adults is about 4-100%. thymus, which is an important central immune organ and an important place for the development of T lymphocytes to leave the bone marrow. The thymus provides a specific microenvironment for the development of T lymphocytes, and the thymus microenvironment. Research on the development, differentiation and maturation of T cells in the thymus is an important area of immunology research today. Studies have shown that T cells express TSHreceptor (TSH-R), and the changes in the development of T cells in the thymus gland have not been fully elucidated. The purpose of this study was to elucidate the thymus T thin in the subsubsubtract mice model. The effect of cell development, the cell composition of the thymus in the SCH mouse model and the changes of T cell apoptosis in different developmental stages. Research methods: Construction of 1.SCH mice model: using methimidazole to feed mice, a dose of methimidazole at a dose of 0.08mg/kg/d, and 16 weeks after a mercapto methimidazole feeding, using I125 labelled radioimmunoassay to detect small amounts. The serum level of free thyroxine (FT4) in the rat serum was tested by Elisa method to test the level of TSH in the serum of mice. By evaluating the thyroid function of the mice, the SCH mouse model was constructed to make its serum test conform to the SCH diagnostic standard. The serum TSH level was elevated and the thyroid hormone level was not significantly changed. The effect of TSH on the development of T cells was studied. The analysis of the immunological composition of T in the thymus of.2.SCH mice: detection of the proportion of T cell subgroups in the different developmental stages of T cells in the thymus gland by multi color flow analysis. The ratio of apoptosis in the different developmental stages of the.3. thymus of the.3. thymus was measured: the apoptosis ratio of each T cell subgroup was detected by the apoptotic test kit. 4. the detection of apoptosis related proteins in the thymus of mice: the Western Blot method was used to test the protein level of p-ERK/ERK in the thymus, and the short-term output function of.5. thymus was detected: the spleen DNA in mice was extracted and the TREC level in the T cell DNA of the spleen was detected by PCR in real time. The result: the construction of 1.SCH mouse model: methimidazole feeding mice for 16 weeks, SCH group mice. There was no significant difference in serum FT4 values compared with the control group (P0.05); the serum level of TSH in the SCH model group was significantly higher than that of the control group (P0.05). The difference in serum TSH level between the two groups was statistically significant in the immunological analysis of T in the different developmental stages of the thymus of.2.SCH mice: the thymus weight of SCH mice was more than the 18%. thymus in the control group. CD4 + and CD8 + single positive (single positive, SP) thymic cells increased the percentage of apoptotic T cells in 38% and 44%.3. thymus, respectively. By detecting apoptosis, we found that the proportion of Annexin-V+ cells in the thymus of SCH mice decreased (P0.05), which showed that TSH decreased the apoptosis phase in the thymus gland of SCH mice. The Western blot results showed that the p-ERK 1/2 in the thymus of SCH mice increased, indicating that the ERK pathway in the thymus of the SCH mice was activated to detect the short-term output function of the.5. thymus: PCR detection of the T cell receptor excision ring in the spleen tissues of mice (TREC). The results showed that TSH promoted the development of thymus T cells in SCH mice and enhanced the recent output of thymus. The mechanism may be through inhibiting the apoptosis of thymus cells, and the anti apoptosis effect may be through the p-ERK/ERK signaling pathway.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R581.2

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