本文關(guān)鍵詞： 陽虛體質(zhì) 陰虛體質(zhì) 唾液 miRNA 中醫(yī)體質(zhì)學　出處：《北京中醫(yī)藥大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:1.探索陽虛、陰虛體質(zhì)唾液miRNA基因表達譜微觀差異,篩選兩種偏頗體質(zhì)微觀可分的標志物,探索體質(zhì)客觀研究的新方向。2.分析兩種偏頗體質(zhì)與平和質(zhì)唾液miRNA基因表達譜微觀差異,為下一步通過調(diào)體逆轉(zhuǎn)非健康狀態(tài)體質(zhì)人群以及診斷和預防疾病提供新的切入點。方法:1.本實驗在一般人群中共招募平和體質(zhì)狀態(tài)者5人,陽虛體質(zhì)組未病狀態(tài)者5例、陰虛體質(zhì)組未病狀態(tài)者6例,共計16名不同職業(yè)、不同年齡的志愿者。用無菌水漱口以清除口腔內(nèi)干擾異物,收集待檢人員口腔內(nèi)唾液。分離口腔唾液并提取其中的RNA,進行HumanmiRNAOneArray高通量基因芯片檢測。2.通過對miRNA表達譜數(shù)據(jù)進行分析,篩選出具有差異性表達的miRNA。3.對差異表達miRNA進行靶基因分析,并對靶基因進行功能富集分析。結(jié)果:1.本研究發(fā)現(xiàn)陽虛體質(zhì)組與平和體質(zhì)組相比共有81個差異表達miRNA,包括33個上調(diào)差異miRNA和48個下調(diào)差異miRNA;陰虛體質(zhì)組與平和體質(zhì)組相比共有98個差異表達miRNA,包括35個上調(diào)差異miRNA和63個下調(diào)差異miRNA;陰虛體質(zhì)與陽虛體質(zhì)相比共有167個差異表達miRNA包括51個上調(diào)差異miRNA和116個下調(diào)差異miRNA。2.通過對差異表達miRNA的靶基因進行KEGG和GO富集分析發(fā)現(xiàn):陽虛與平和對比組存在激素代謝、糖脂代謝、免疫調(diào)節(jié)、腫瘤等生物學通路的異常表達;陰虛與平和對比組存在炎癥反應、免疫調(diào)節(jié)、腫瘤等生物學通路的異常表達;陽虛與陰虛對比組存在免疫調(diào)節(jié)、激素代謝、腫瘤等生物學通路的異常表達。結(jié)論:1.陽虛與平和、陰虛與平和、陽虛與陰虛組在全基因組層面存在系統(tǒng)性的miRNA表達譜的差異。2.通過靶基因KEGG分析發(fā)現(xiàn)陽虛、陰虛體質(zhì)對比平和質(zhì)組上調(diào)靶基因富集通路中均存在免疫通路的差異表達,顯示兩種偏頗體質(zhì)在基因?qū)用婵赡芫嬖诿庖邔W通路異常的分子機制。與此同時陽虛對比平和組甲狀腺激素、雄性激素等相關(guān)激素通路的差異表達,陰虛對比平和組炎性通路、激素相關(guān)通路的差異表達,可能是導致陽虛體質(zhì)虛性低靡特征和陰虛體質(zhì)虛性亢奮特征的內(nèi)在原因。3.本研究以人體口腔唾液作為研究對象,選用miRNA表達譜分析作為研究方式,初步構(gòu)建了陰虛陽虛體質(zhì)的唾液miRNA表達譜,篩選差異目的基因,分析KEGG、GO富集結(jié)果,為探索陽虛、陰虛兩種偏頗體質(zhì)異常表現(xiàn)的分子機制奠定了基礎(chǔ)。于此同時將人體口腔唾液作為研究的出發(fā)點,也是為了探尋一種更簡便可行的方式,為體質(zhì)微觀辨識探尋新方向。
[Abstract]:Objective 1. To explore the microcosmic difference of saliva miRNA gene expression profile between yang-deficiency and yin-deficiency constitution, and to screen two kinds of microcosmic markers of biased constitution. To explore the new direction of objective study of physique. 2. To analyze the microcosmic difference of miRNA gene expression profile between two kinds of biased constitution and placid saliva. This study provides a new entry point for the next step to reverse the unhealthy physique population and to diagnose and prevent diseases. Methods: 1. In the general population, 5 healthy persons were recruited in this experiment, and 5 cases in the Yang deficiency body group were not ill. In Yin-deficiency constitution group, 6 cases were not ill, a total of 16 volunteers of different occupations and ages. Gargle with aseptic water was used to remove foreign body interference in oral cavity. The oral saliva was collected, the oral saliva was isolated and the RNAs were extracted from the saliva, and the HumanmiRNAOneArray high-throughput gene chip was used to detect .2. the data of miRNA expression profile were analyzed. The differentially expressed miRNA.3.The target gene of differentially expressed miRNA was analyzed. The functional enrichment analysis of target gene was carried out. Results 1. This study found that there were 81 differentially expressed miRNAs in Yang deficiency constitution group compared with peaceful constitution group, including 33 up-regulated differential miRNA and 48 down-regulated miRNAs; Yin deficiency constitution group and calmer body group. There were 98 differentially expressed miRNAs, including 35 up-regulated miRNA and 63 down-regulated miRNAs, and 167 differentially expressed miRNA including 51 up-regulated differential miRNA and 116 down-regulated differential miRNA.2. KEGG and go enrichment analysis of target genes expressing differentially expressed miRNA showed that hormone metabolism existed in the control group of Yang deficiency and Ping Ping. Abnormal expression of biological pathways such as glucose and lipid metabolism, immunomodulation, tumor and so on; abnormal expression of inflammatory response, immune regulation, tumor and other biological pathways in Yin-deficiency and calming control groups; immune regulation in Yang deficiency and Yin-deficiency control groups. Conclusion: 1. Yang deficiency and peace, yin deficiency and peace, yang deficiency and yin deficiency have systematic difference of miRNA expression profile on the whole genome level. 2. Yang deficiency is found by KEGG analysis of target gene. There was differential expression of immune pathway in the up-regulated target gene enrichment pathway in calming group compared with that in Yin deficiency group. At the same time, the differential expression of thyroid hormone, androgen and other related hormone pathways in Yang deficiency contrast and Ping Ping group, and the inflammatory pathway in Yin deficiency contrast peace group. The differential expression of hormone related pathways may be the internal cause of the low prevalence of deficiency of Yang and the hyperactivity of deficiency of yin. 3. In this study, oral saliva was used as the research object, and miRNA expression profile analysis was used as the research method. The saliva miRNA expression profile of yin deficiency and yang deficiency constitution was preliminarily constructed, the differentially expressed target genes were screened, and the enrichment results of KEGG go were analyzed in order to explore yang deficiency. The molecular mechanism of two kinds of biased physical abnormalities of yin deficiency has laid a foundation. At the same time taking oral saliva as the starting point is also to explore a more convenient and feasible way to explore a new direction for physique microscopic identification.
【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R229

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