本文選題：橋本甲狀腺炎 + IFN-γ��； 參考：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的1.探討橋本甲狀腺炎(Hashimoto thyroiditis,HT)甲狀腺組織Th1細(xì)胞因子γ-干擾素(interferon-γ,IFN-γ)與凋亡蛋白Fas之間的關(guān)系。2.探討Th1細(xì)胞因子IFN-γ對(duì)大鼠甲狀腺細(xì)胞系(FRTL-5細(xì)胞)Fas表達(dá)的調(diào)控。方法1.橋本甲狀腺炎甲狀腺組織來源于甲狀腺病理組織切片,收集甲狀腺腺瘤旁正常甲狀腺組織作為正常對(duì)照組;采用免疫組織化學(xué)方法檢測(cè)HT和正常甲狀腺組織中IFN-γ及Fas的表達(dá)和分布。2.對(duì)體外培養(yǎng)的FRTL-5細(xì)胞,采用兩種干預(yù)方法:(1)使用終濃度為40 ng/ml的IFN-γ分別干預(yù)FRTL-5細(xì)胞6、12、24、48、72h為IFN-γ刺激組,未經(jīng)IFN-γ作用的FRTL-5細(xì)胞為對(duì)照組,收集細(xì)胞用于后期檢測(cè);(2)終濃度分別為5、10、20、40、80 ng/ml的IFN-γ干預(yù)FRTL-5細(xì)胞為IFN-γ刺激組,未經(jīng)IFN-γ作用的FRTL-5細(xì)胞為對(duì)照組,培養(yǎng)48h后收集細(xì)胞用于后期檢測(cè)。3.使用TRIzol提取FRTL-5細(xì)胞總RNA;逆轉(zhuǎn)錄試劑盒合成c DNA。應(yīng)用Real-Time PCR檢測(cè)各組細(xì)胞Fas m RNA的表達(dá)水平,基因表達(dá)量用2-ΔΔCt相對(duì)量表示。4.采用FACSVerse流式細(xì)胞儀,分析在不同濃度IFN-γ干預(yù)條件下FRTL-5細(xì)胞Fas蛋白表達(dá)。結(jié)果1.正常甲狀腺組織內(nèi)IFN-γ表達(dá)陰性,Fas陽(yáng)性表達(dá)甲狀腺濾泡細(xì)胞不明顯。在被檢的HT患者組織切片中,均見到浸潤(rùn)的淋巴細(xì)胞IFN-γ表達(dá)陽(yáng)性,甲狀腺濾泡細(xì)胞Fas表達(dá)陽(yáng)性,且Fas蛋白陽(yáng)性甲狀腺濾泡細(xì)胞常見于淋巴細(xì)胞浸潤(rùn)區(qū),無淋巴細(xì)胞浸潤(rùn)的區(qū)域,較少見到甲狀腺濾泡細(xì)胞Fas表達(dá)陽(yáng)性。2.IFN-γ上調(diào)FRTL-5細(xì)胞Fas m RNA表達(dá),且存在時(shí)間-劑量依賴關(guān)系。FRTL-5細(xì)胞Fas m RNA的表達(dá)量與IFN-γ刺激時(shí)間相關(guān),24h、48h、72h組皆顯著高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(`x±s分別為6.39±2.02,7.45±4.33,11.40±5.05,P0.01);FRTL-5細(xì)胞Fas m RNA的表達(dá)量與IFN-γ刺激濃度相關(guān),20ng/ml、40ng/ml、80ng/ml組皆顯著高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(`x±s分別為4.01±1.48,10.53±1.54,3.62±2.22,P0.01)。3.流式細(xì)胞術(shù)檢測(cè)示未經(jīng)IFN-γ刺激的FRTL-5細(xì)胞Fas蛋白陽(yáng)性表達(dá)率為10.1%,隨著IFN-γ的劑量的增加,FRTL-5細(xì)胞Fas蛋白陽(yáng)性表達(dá)率逐漸增高,20ng/ml的IFN-γ刺激組FRTL-5細(xì)胞Fas蛋白陽(yáng)性表達(dá)率為28.3%,40ng/ml刺激組Fas蛋白陽(yáng)性表達(dá)率為30.0%,80ng/ml刺激組Fas蛋白陽(yáng)性表達(dá)率為38.9%。結(jié)論:橋本甲狀腺炎甲狀腺組織浸潤(rùn)的淋巴細(xì)胞分泌的IFN-γ通過上調(diào)甲狀腺細(xì)胞Fas的表達(dá),導(dǎo)致甲狀腺組織的凋亡,與HT患者的甲狀腺功能減退可能相關(guān)。
[Abstract]:Objective 1.To investigate the relationship between Th1 cytokine interferon- 緯 (IFN- 緯) and apoptotic protein (Fas) in thyroid tissue of Hashimoto thyroiditis (HT).To investigate the effect of Th1 cytokine IFN- 緯 on the expression of FAS in rat thyroid cell line FRTL-5.Method 1.The thyroid tissue of Hashimoto's thyroiditis originated from the pathological section of thyroid gland and the normal thyroid tissue adjacent to thyroid adenoma was collected as the normal control group.Immunohistochemical method was used to detect the expression and distribution of IFN- 緯 and Fas in HT and normal thyroid tissues.FRTL-5 cells cultured in vitro were treated with IFN- 緯 at the final concentration of 40 ng/ml for 72 h, and FRTL-5 cells without IFN- 緯 were treated with IFN- 緯 for 72 h.After 48 hours of culture, the cells collected were treated with IFN- 緯 as IFN- 緯 stimulation group and FRTL-5 cells without IFN- 緯 as control group. After 48 hours of culture, the cells were collected for later detection.Total RNAs of FRTL-5 cells were extracted by TRIzol and c DNA was synthesized by reverse transcription kit.The expression level of Fas m RNA was detected by Real-Time PCR, and the expression of Fas m RNA was expressed by the relative amount of 2- 螖 Ct.FACSVerse flow cytometry was used to analyze the expression of Fas protein in FRTL-5 cells treated with different concentrations of IFN- 緯.Result 1.The positive expression of IFN- 緯 in thyroid follicular cells was not obvious.The positive expression of IFN- 緯 in infiltrated lymphocytes and Fas expression in thyroid follicular cells were found in the tissue sections of HT patients. The positive expression of Fas protein in thyroid follicular cells was found in the lymphocytic infiltrating area, without lymphocytic infiltration.The positive expression of Fas in thyroid follicular cells. 2. IFN- 緯 upregulated the expression of Fas m RNA in FRTL-5 cells, and the expression of Fas m RNA in FRTL-5 cells was significantly higher than that in the control group for 72 h, and the expression of Fas m RNA in FRTL-5 cells was significantly higher than that in the control group.The expression of Fas m RNA in FRTL-5 cells was significantly higher than that in the control group ('x 鹵s 6.39 鹵2.02 鹵7.45 鹵4.33 鹵11.40 鹵5.05 鹵5.05), respectively. The expression of Fas m RNA in FRTL-5 cells was significantly higher than that in the control group (`x 鹵s = 4.01 鹵1.48U 10.53 鹵1.543.62 鹵2.22g / ml P 0.01g / ml), and was significantly higher than that in the control group (`x 鹵s = 4.01 鹵1.48U 10.53 鹵1.543.62 鹵2.22g / ml P 0.01g / ml).Flow cytometry showed that the positive expression rate of Fas protein in FRTL-5 cells without IFN- 緯 stimulation was 10.1. With the increase of the dose of IFN- 緯, the positive expression rate of Fas protein in FRTL-5 cells gradually increased with 20 ng / ml IFN- 緯 stimulation. The positive expression rate of Fas protein in FRTL-5 cells was 28.3ng / ml.The positive expression rate of Fas protein in the stimulation group was 30.0% and the positive expression rate of Fas protein was 38.9% in the 80ng / ml stimulation group.Conclusion: IFN- 緯 secreted by infiltrated lymphocytes in Hashimoto thyroiditis leads to thyroid apoptosis by up-regulating the expression of Fas in thyroid cells, which may be related to hypothyroidism in HT patients.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R581.4

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