發(fā)布時(shí)間：2018-11-16 14:57

【摘要】：背景干擾素-α(interferon-α,IFN-α)是經(jīng)典的固有免疫細(xì)胞因子,具有顯著的抗病毒和抗腫瘤作用,因此在臨床上IFN-α常用于抗病毒感染(丙型肝炎、乙型肝炎)或抗腫瘤(如黑色素瘤、乳腺癌)治療。IFN-α的長(zhǎng)期使用可以導(dǎo)致30～70%HCV患者出現(xiàn)不同程度的精神抑郁。這給患者帶來(lái)額外的痛苦。因此很有必要尋找一種有效的方法,及時(shí)監(jiān)測(cè)接受IFN-α治療患者可在治療中的抑郁發(fā)病風(fēng)險(xiǎn),以便及時(shí)采取相應(yīng)的治療措施,如降低IFN-α用藥量,改變用藥頻次或者施加抗抑郁治療,避免患者出現(xiàn)抑郁癥狀。目的尋找一種客觀而準(zhǔn)確的預(yù)測(cè)抑郁發(fā)病風(fēng)險(xiǎn)的生物學(xué)標(biāo)志(易感基因)。方法1.給藥：在SH-sy5y細(xì)胞培養(yǎng)液中加入終濃度為3000IU/mL的干擾素-α(IFN-α),持續(xù)16h,同時(shí)設(shè)PBS對(duì)照組。2. mRNA提�。菏褂肨rizol法提取SH-SY5Y細(xì)胞總RNA。3.逆轉(zhuǎn)錄為cDNA:將提取到的SH-SY5Y細(xì)胞總RNA逆轉(zhuǎn)錄為cDNA。4.基因芯片檢測(cè)：分別檢測(cè)5-羥色胺、多巴胺代謝相關(guān)基因、炎癥相關(guān)基因、神經(jīng)生長(zhǎng)相關(guān)基因、經(jīng)典的抑郁相關(guān)基因的表達(dá)。實(shí)驗(yàn)重復(fù)三次。5.統(tǒng)計(jì)分析：使用spss19.0統(tǒng)計(jì)軟件對(duì)數(shù)據(jù)進(jìn)行T檢驗(yàn),P0.05為顯著差異。抑郁易感基因的篩選條件為：IFN-α處理組比PBS對(duì)照組的比值2.0或者0.5,并且P0.05。結(jié)果1.全基因表達(dá)譜整張芯片信號(hào)均勻、清晰；沒(méi)有影響信號(hào)值的邊緣化效應(yīng)或刮傷等；陽(yáng)性定位點(diǎn)和陰性對(duì)照區(qū)結(jié)果明顯。2.全基因表達(dá)譜芯片聚類圖顯示每組3次重復(fù)實(shí)驗(yàn)的樣品聚類到一起,證明實(shí)驗(yàn)重復(fù)性好。3. IFN-α能夠提高SH-SY5Y細(xì)胞TPH2、CASP1、INDO、IL7、IL12A、IL15 RNA的表達(dá)水平。結(jié)論IFN-α能夠提高SH-SY5Y細(xì)胞TPH2、CASP1、INDO、IL7、IL12A、IL15RNA的表達(dá)水平。
[Abstract]:Background interferon- 偽 (IFN- 偽) is a classical innate immune cytokine with significant antiviral and anti-tumor effects. Therefore, IFN- 偽 is often used in clinical antiviral infection (hepatitis C). Hepatitis B) or antitumor therapy (such as melanoma, breast cancer). Long-term use of IFN- 偽 can lead to varying degrees of depression in patients with 30~70%HCV. This brings additional pain to the patient. Therefore, it is necessary to find an effective method to monitor the risk of depression in patients treated with IFN- 偽 in time, so as to take corresponding treatment measures, such as reducing the dosage of IFN- 偽. Change the frequency of medication or apply antidepressant treatment to avoid depressive symptoms. Objective to find an objective and accurate biomarker (susceptibility gene) for predicting the risk of depression. Method 1. Administration: the final concentration of 3000IU/mL interferon-偽 (IFN- 偽) was added to the SH-sy5y cell culture medium for 16 h, and the PBS control group was set up at the same time. MRNA extraction: total RNA.3. extraction from SH-SY5Y cells by Trizol Reverse transcription to cDNA: the total RNA of extracted SH-SY5Y cells is reversed to cDNA.4. Gene chip detection: 5-hydroxytryptamine, dopamine metabolism related genes, inflammatory genes, nerve growth related genes, classic depression related genes were detected. The experiment was repeated three times. Statistical analysis: using spss19.0 statistical software for T test data, P0.05 is the significant difference. The screening conditions of depression susceptibility genes were as follows: the ratio of IFN- 偽 treatment group to PBS control group was 2.0 or 0.5, and P 0.05. Result 1. The whole gene expression profile was uniform and clear, there was no marginal effect or scrape, the results of positive locus and negative control were obvious. 2. The whole gene expression microarray cluster diagram showed that the samples of each group of three repeated experiments were clustered together, which proved that the experiment was reproducible. IFN- 偽 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15 RNA in SH-SY5Y cells. Conclusion IFN- 偽 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15RNA in SH-SY5Y cells.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R96

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