IFN-α对人神经细胞系SH-SY5Y基因表达谱的影响
发布时间：2018-11-16 14:57
【摘要】：背景干扰素-α(interferon-α,IFN-α)是经典的固有免疫细胞因子,具有显著的抗病毒和抗肿瘤作用,因此在临床上IFN-α常用于抗病毒感染(丙型肝炎、乙型肝炎)或抗肿瘤(如黑色素瘤、乳腺癌)治疗。IFN-α的长期使用可以导致30～70%HCV患者出现不同程度的精神抑郁。这给患者带来额外的痛苦。因此很有必要寻找一种有效的方法,及时监测接受IFN-α治疗患者可在治疗中的抑郁发病风险,以便及时采取相应的治疗措施,如降低IFN-α用药量,改变用药频次或者施加抗抑郁治疗,避免患者出现抑郁症状。目的寻找一种客观而准确的预测抑郁发病风险的生物学标志(易感基因)。方法1.给药：在SH-sy5y细胞培养液中加入终浓度为3000IU/mL的干扰素-α(IFN-α),持续16h,同时设PBS对照组。2. mRNA提取：使用Trizol法提取SH-SY5Y细胞总RNA。3.逆转录为cDNA:将提取到的SH-SY5Y细胞总RNA逆转录为cDNA。4.基因芯片检测：分别检测5-羟色胺、多巴胺代谢相关基因、炎症相关基因、神经生长相关基因、经典的抑郁相关基因的表达。实验重复三次。5.统计分析：使用spss19.0统计软件对数据进行T检验,P0.05为显著差异。抑郁易感基因的筛选条件为：IFN-α处理组比PBS对照组的比值2.0或者0.5,并且P0.05。结果1.全基因表达谱整张芯片信号均匀、清晰；没有影响信号值的边缘化效应或刮伤等；阳性定位点和阴性对照区结果明显。2.全基因表达谱芯片聚类图显示每组3次重复实验的样品聚类到一起,证明实验重复性好。3. IFN-α能够提高SH-SY5Y细胞TPH2、CASP1、INDO、IL7、IL12A、IL15 RNA的表达水平。结论IFN-α能够提高SH-SY5Y细胞TPH2、CASP1、INDO、IL7、IL12A、IL15RNA的表达水平。
[Abstract]:Background interferon- 伪 (IFN- 伪) is a classical innate immune cytokine with significant antiviral and anti-tumor effects. Therefore, IFN- 伪 is often used in clinical antiviral infection (hepatitis C). Hepatitis B) or antitumor therapy (such as melanoma, breast cancer). Long-term use of IFN- 伪 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- 伪 in time, so as to take corresponding treatment measures, such as reducing the dosage of IFN- 伪. 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-伪 (IFN- 伪) 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- 伪 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- 伪 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15 RNA in SH-SY5Y cells. Conclusion IFN- 伪 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15RNA in SH-SY5Y cells.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R96
[Abstract]:Background interferon- 伪 (IFN- 伪) is a classical innate immune cytokine with significant antiviral and anti-tumor effects. Therefore, IFN- 伪 is often used in clinical antiviral infection (hepatitis C). Hepatitis B) or antitumor therapy (such as melanoma, breast cancer). Long-term use of IFN- 伪 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- 伪 in time, so as to take corresponding treatment measures, such as reducing the dosage of IFN- 伪. 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-伪 (IFN- 伪) 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- 伪 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- 伪 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15 RNA in SH-SY5Y cells. Conclusion IFN- 伪 can increase the expression of TPH2,CASP1,INDO,IL7,IL12A,IL15RNA in SH-SY5Y cells.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R96
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