基于“先天伏寒”病因所致“先天伏寒证”人的血清蛋白组学研究

发布时间：2018-04-09 17:54

本文选题：先天伏寒证　切入点：肾阳虚证　出处：《长春中医药大学》2015年博士论文

【摘要】：目的:本研究拟采用蛋白组学研究方法,对健康人、先天伏寒证、肾阳虚证以及辩证治疗后的先天伏寒证人群的血清样本进行蛋白组学分析,寻找差异表达的蛋白质,结合生物信息学和统计学对差异蛋白分析,探索先天伏寒证、肾阳虚证与蛋白质之间的关系,建立一个“先天伏寒证候-蛋白质表达谱”,并对图谱上有意义的蛋白进行质谱鉴定,并根据筛选到的潜在证候标记物,寻找“先天伏寒”证的证候标记物。方法:应用病证结合的方法,采用二维凝胶电泳和生物质谱结合的技术对先天伏寒证、肾阳虚证与健康人及先天伏寒证治疗前后的差异蛋白点进行分离和鉴定,从分子生物学角度研究亚健康状态下先天伏寒证的分子实质,以期发现先天伏寒证的物质基础。结果:1.通过蛋白质组学方法得到先天伏寒证和阳虚证患者间差异蛋白点36个,其中表达上调蛋白20个,表达下调蛋白16个;伏寒组与健康人有差异蛋白点28个,其中表达上调蛋白9个,表达下调蛋白19个;阳虚组与健康人有差异蛋白点28个,其中表达上调蛋白有15个,表达下调蛋白13个;通过质谱鉴定分析,先天伏寒证与阳虚证二者有一些共性又有差异蛋白点。2、伏寒组与健康人血清蛋白表达比较,经鉴定筛选后上调的蛋白质主要有补体C4前体蛋白原和角蛋白,下调的蛋白质主要有补体因子H、补体C6、补体因子I,胆绿素还原酶,维生素D结合蛋白,血红素结合蛋白,血浆丝氨酸蛋白酶抑制剂,人体绒膜促性腺激素,激肽原等。而伏寒组经药物干预后,补体因子C4水平与角蛋白水平均下调。胆绿素还原酶,血红素结合蛋白,维生素D结合蛋白等均上调3、阳虚组与健康人血清蛋白表达比较,上调的蛋白质有糖蛋白,胆绿素还原酶,维生素D,丛生蛋白,角蛋白,过氧化氢酶抑制剂,碳酸酐酶,而下调的蛋白质有补体因子C4,血液结合素,羧肽酶催化剂。4、从伏寒与健康人表达的差异蛋白及阳虚与健康人表达差异的蛋白比较来分析,二者有一些共性或有交叉的蛋白点,其中角蛋白与健康人比较均是上调的,血红素结合蛋白均是下调的。而胆绿素还原酶、维生素D结合蛋白在伏寒组人群中表达是下调的,而在阳虚组表达是上调的。补体C4在伏寒组人群中表达是上调的,而在阳虚组表达是下调的。结论:无论是伏寒组还是阳虚组,通过与健康人血清蛋白比较均存在明显差异,说明两者都处于非健康状态。伏寒组与阳虚组在血清蛋白表达上也是具有明显差异,说明二者证候是不同的。阳虚组病更多地表现为糖蛋白的表达异常,伏寒组更多的是体现在补体系统的异常。伏寒组经药物干预后,部分与人体免疫、动脉硬化相关的蛋白可以得到调整,说明早期识别及早期干预对疾病的预防具有重大意义。
[Abstract]:Objective: the aim of this study was to analyze the serum samples of healthy people, congenital latent cold syndrome, kidney-yang deficiency syndrome and after dialectical treatment with proteomics, and to find the differentially expressed proteins.Combining with bioinformatics and statistics to analyze the differential protein, to explore the relationship between the congenital latent cold syndrome, the kidney yang deficiency syndrome and the protein, and to establish a "congenital latent cold syndromes-protein expression profile".The significant proteins on the map were identified by mass spectrometry, and the syndrome markers of "congenital latent cold" syndrome were found according to the potential syndrome markers screened.Methods: two dimensional gel electrophoresis and biological mass spectrometry were used to separate and identify the differential protein points of congenital cold accumbent syndrome, kidney yang deficiency syndrome and healthy persons and congenital latent cold syndrome before and after treatment.In order to find out the material basis of congenital latent cold syndrome, the molecular essence of congenital latent cold syndrome under sub-health condition was studied from the molecular biological point of view.The result is 1: 1.A total of 36 differential protein points were obtained by proteomics, including 20 up-regulated proteins and 16 down-regulated proteins, and 28 differentially expressed protein points were found between the latent cold group and healthy persons.Among them, 9 were up-regulated proteins, 19 were down-regulated proteins, 28 were differentially expressed between Yang deficiency group and healthy people, among which 15 were up-regulated proteins and 13 were down-regulated proteins.The expression of serum protein in cold latent group was compared with that in healthy people. The up-regulated protein mainly included complement C4 precursor protein and keratin after identification and screening.The down-regulated proteins mainly include complement factor H, complement C6, complement factor I, cholgreen reductase, vitamin D binding protein, heme binding protein, plasma serine protease inhibitor, human chorionic gonadotropin, kallikinogen and so on.However, the level of complement factor C4 and keratin decreased after drug intervention in latent cold group.The expression of serum protein in Yang deficiency group was compared with that in healthy people. The up-regulated proteins included glycoprotein, cholgreen reductase, vitamin D, cluster protein, keratin, etc.Catalase inhibitors, carbonic anhydrase, and down-regulated proteins include complement factor C4, blood binding factor, carboxypeptidase catalyst.There are some common or cross protein spots between them. Keratin is up-regulated and heme binding protein is down-regulated compared with healthy people.However, the expression of bile green reductase and vitamin D binding protein was down-regulated in cold cold group and up-regulated in yang deficiency group.The expression of complement C4 was up-regulated in cold cold group and down-regulated in yang deficiency group.Conclusion: there are significant differences in serum protein between cold latent group and yang deficiency group, indicating that both groups are in unhealthy state.There was also significant difference in the expression of serum protein between cold latent group and yang deficiency group, indicating that the syndromes were different between them.The abnormal expression of glycoprotein in Yang deficiency group was more than that in cold latent group.After drug intervention, some proteins related to human immunity and atherosclerosis could be adjusted, which indicated that early recognition and early intervention were of great significance to the prevention of diseases.
【学位授予单位】：长春中医药大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R228

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