儿童过敏性紫癜基因表达谱及肾损伤相关基因研究

发布时间：2018-04-06 21:26

本文选题：过敏性紫癜　切入点：紫癜性肾炎　出处：《南昌大学》2012年硕士论文

【摘要】：目的：用Roche NimbleGen人基因表达谱芯片技术筛选过敏性紫癜(HenochSchonlein purpura，HSP)患儿疾病发生及肾损伤相关基因，以进一步从分子水平更全面、系统地分析儿童HSP发生发展及肾损伤发生机制。方法：利用Roche NimbleGen人基因表达谱芯片（约含有45033个基因）,分别检测单纯过敏性紫癜组外周血（HSP）、紫癜性肾炎组外周血(HSPN)、健康对照组外周血（C）的基因表达谱，用相关生物信息学分析方法对不同实验组间基因表达谱进行差异分析，最后应用实时定量PCR技术对部分差异表达基因进行验证。结果： 1、单纯过敏性紫癜组与对照组间：共有1064条差异表达基因，其中2倍以上差异基因225条，2.5倍以上38条，3倍以上5条；其中225条2倍以上差异基因中146条表达上调，79条表达下调。紫癜性肾炎组与单纯过敏性紫癜组：共有830条差异表达基因，其中2倍以上差异基因283条，2.5倍以上118条，3倍以上51条；其中283条2倍以上差异基因中180条表达上调，103条表达下调。 2、随机选取其中3条差异基因进行Real．time PCR验证，其结果与基因芯片筛查结果一致，结果充分证实表达谱芯片差异表达基因结果的可靠性。 3、对单纯过敏性紫癜组与对照组2倍以上差异基因进行GO生物学过程分析发现，已知功能分类的共119条，主要涉及：转录/翻译、物质合成/代谢过程和信号转导/通路相关基因，其次为分化/发育、免疫/炎症和凋亡/抗凋亡相关基因；而在紫癜性肾炎组与单纯过敏性紫癜组2倍以上差异基因中，我们发现已知功能分类的有201条，主要涉及：物质合成/代谢过程相关基因，其次为转录/翻译、免疫/炎症、信号转导/通路、分化/发育和细胞周期。 4、初步筛选出免疫炎症/细胞黏附/信号转导通路相关基因AIF1、LAMC2、BCAM、CD209、MFAP4、ELMOD3、SCUBE1、ADAM33、RET、CLEC7A、MASP2、TAPS、AOAH、MGST2、PSMA3、SP100，代谢/凋亡相关基因MUC3B、MUC1、HSD11B1、MTHFS、TNFAIP8及未知基因BM88、CEACAM3等可能在HSP发病中起重要作用，，同样也筛选出免疫炎症/信号转导通路相关基因KRT18、GRB10、VEGFB、HSP90AB1、HSPD1、F12、TGFBR2、TGM2、UBE2L6、CTSS、PXDN、GBP2、HLA-DRB1、HLA-DMA、HLA-DPA1，代谢/分化发育/凋亡相关基因SORD、HSPE1、ARG2、HSP90AA2、PRAME、TNFAIP2、MMP19、ZFP36L1，细胞黏附相关基因CLDN4及未知基因CKMT1B等可能与HSP肾脏损伤的发生密切相关。结论： 1、首次用快速高通量的表达谱芯片筛选出过敏性紫癜及紫癜性肾炎相关基因。 2、初步筛选出(AIF1、BCAM、CLEC7A、CD209、AOAH、MAPS2等)可能导致HSP发病的易感基因，进一步证实了免疫炎症、细胞黏附相关基因的表达异常可能在HSP发病过程中起到了主导作用，同时也发现了一些可能导致HSP发生的代谢/凋亡相关基因（如MUC3B、HSD11B1、MTHFS、TNFAIP8等）及未知基因。 3、研究不仅证实了在HSP肾损伤过程中可能起重要作用的已被认识的基因如VEGFB、HLA-DRB1，也首次发现了一些以往在HSPN发病过程中未曾报道的已知基因（如SORD、ARG2、ZFP36L1、TGM2、KRT18等）以及未知基因。 4、针对这些基因的深入研究，将有助于更全面、系统地揭示过敏性紫癜的分子机制。
[Abstract]:Purpose :

The pathogenesis of HSP and the pathogenesis of renal injury in children with Henoch Schonlein purpura ( HSP ) were screened by using the Roche NileGen gene expression profiling chip technique .

Method :

The gene expression profiles of peripheral blood ( HSP ) , Henoch - Schonlein purpura nephritis ( HSPN ) and healthy control group ( C ) were detected by using the Roche NileGen gene expression profiling chip ( about 45033 genes ) . The gene expression profiles of different experimental groups were analyzed by means of bioinformatics analysis . Finally , the partial differential expression gene was verified by real - time quantitative PCR .

Results :

1 . There were a total of 1064 differential expression genes , among which more than 2 times the difference gene was 225 , 2.5 times more than 38 , 3 times more than 5 ;
Among them , more than 225 of these genes were up - regulated and 79 were down - regulated . There were 830 differentially expressed genes , among them , there were 830 differentially expressed genes , more than 2 times the difference gene 283 , 2.5 - fold or more than 118 , and more than 3 times 51 ;
Of these , the expression of 180 in 283 2 - fold difference gene was up - regulated and 103 expression was down - regulated .

2 . Three different genes were randomly selected for Real . time PCR . The results were consistent with the results of gene chip screening .

3 . The analysis of GO biological process of 2 - fold difference gene in Henoch - Henoch - Schonlein purpura group and control group showed that 119 of the known functions were involved : transcription / translation , substance synthesis / metabolic process and signal transduction / pathway - related gene , followed by differentiation / development , immune / inflammation and apoptosis / anti - apoptosis related genes ;
Among the 2 - fold difference genes in the Henoch - Schonlein purpura nephritis group and the simple allergic purpura group , we found that there were 201 known functional classes , mainly involved in gene synthesis / metabolic process - related genes , followed by transcription / translation , immune / inflammation , signal transduction / pathway , differentiation / development and cell cycle .

4 . The related genes AIF1 , LAMC2 , BCAM , CD209 , MFAP4 , ELMOD3 , SCUBE1 , CD209 , MAP4 , ELMOD3 , SCUBE1 , TAPS , AOAH , MGST2 , PSMA3 , TAPS , AOAH , MGST2 , PSMA3 , SP100 , TNFAIP8 and unknown genes BM88 , CEACAM3 and so on may play an important role in the pathogenesis of HSP . MMP - 19 , ZFP36L1 , cell adhesion - related gene CLDN4 and unknown gene CKMT1B may be closely related to HSP renal injury .

Conclusion :

1 . The genes associated with Henoch - Schonlein purpura and Henoch - Schonlein purpura nephritis were screened for the first time with a fast high - throughput expression profiling chip .

2 . Preliminary screening ( AIF1 , BCAM , CLEC7A , CD209 , AOAH , MAPS2 , etc . ) may lead to the susceptibility gene of HSP , further confirming the immune inflammation , the abnormal expression of cell adhesion - related genes may play a leading role in the pathogenesis of HSP , and some genes related to the metabolism / apoptosis of HSP ( such as MUC3B , HSD 11B1 , HFS , TNFAIP8 , etc . ) and unknown genes are also found .

3 . The research not only confirmed that the known genes , such as VEGFB , HLA - DRB1 , which may play an important role in the course of HSP kidney injury , also first discovered some known genes ( such as SORD , ARG2 , ZFP36L1 , TGM2 , KRT18 , etc . ) which have not been reported in the pathogenesis of HSP .

4 . In - depth study of these genes will help more comprehensively and systematically reveal the molecular mechanism of Henoch - Schonlein purpura .

【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.5

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