本文選題：先天伏寒證　切入點：腎陽虛證　出處：《長春中醫(yī)藥大學(xué)》2015年博士論文

【摘要】：目的:本研究擬采用蛋白組學(xué)研究方法,對健康人、先天伏寒證、腎陽虛證以及辯證治療后的先天伏寒證人群的血清樣本進行蛋白組學(xué)分析,尋找差異表達的蛋白質(zhì),結(jié)合生物信息學(xué)和統(tǒng)計學(xué)對差異蛋白分析,探索先天伏寒證、腎陽虛證與蛋白質(zhì)之間的關(guān)系,建立一個“先天伏寒證候-蛋白質(zhì)表達譜”,并對圖譜上有意義的蛋白進行質(zhì)譜鑒定,并根據(jù)篩選到的潛在證候標(biāo)記物,尋找“先天伏寒”證的證候標(biāo)記物。方法:應(yīng)用病證結(jié)合的方法,采用二維凝膠電泳和生物質(zhì)譜結(jié)合的技術(shù)對先天伏寒證、腎陽虛證與健康人及先天伏寒證治療前后的差異蛋白點進行分離和鑒定,從分子生物學(xué)角度研究亞健康狀態(tài)下先天伏寒證的分子實質(zhì),以期發(fā)現(xiàn)先天伏寒證的物質(zhì)基礎(chǔ)。結(jié)果:1.通過蛋白質(zhì)組學(xué)方法得到先天伏寒證和陽虛證患者間差異蛋白點36個,其中表達上調(diào)蛋白20個,表達下調(diào)蛋白16個;伏寒組與健康人有差異蛋白點28個,其中表達上調(diào)蛋白9個,表達下調(diào)蛋白19個;陽虛組與健康人有差異蛋白點28個,其中表達上調(diào)蛋白有15個,表達下調(diào)蛋白13個;通過質(zhì)譜鑒定分析,先天伏寒證與陽虛證二者有一些共性又有差異蛋白點。2、伏寒組與健康人血清蛋白表達比較,經(jīng)鑒定篩選后上調(diào)的蛋白質(zhì)主要有補體C4前體蛋白原和角蛋白,下調(diào)的蛋白質(zhì)主要有補體因子H、補體C6、補體因子I,膽綠素還原酶,維生素D結(jié)合蛋白,血紅素結(jié)合蛋白,血漿絲氨酸蛋白酶抑制劑,人體絨膜促性腺激素,激肽原等。而伏寒組經(jīng)藥物干預(yù)后,補體因子C4水平與角蛋白水平均下調(diào)。膽綠素還原酶,血紅素結(jié)合蛋白,維生素D結(jié)合蛋白等均上調(diào)3、陽虛組與健康人血清蛋白表達比較,上調(diào)的蛋白質(zhì)有糖蛋白,膽綠素還原酶,維生素D,叢生蛋白,角蛋白,過氧化氫酶抑制劑,碳酸酐酶,而下調(diào)的蛋白質(zhì)有補體因子C4,血液結(jié)合素,羧肽酶催化劑。4、從伏寒與健康人表達的差異蛋白及陽虛與健康人表達差異的蛋白比較來分析,二者有一些共性或有交叉的蛋白點,其中角蛋白與健康人比較均是上調(diào)的,血紅素結(jié)合蛋白均是下調(diào)的。而膽綠素還原酶、維生素D結(jié)合蛋白在伏寒組人群中表達是下調(diào)的,而在陽虛組表達是上調(diào)的。補體C4在伏寒組人群中表達是上調(diào)的,而在陽虛組表達是下調(diào)的。結(jié)論:無論是伏寒組還是陽虛組,通過與健康人血清蛋白比較均存在明顯差異,說明兩者都處于非健康狀態(tài)。伏寒組與陽虛組在血清蛋白表達上也是具有明顯差異,說明二者證候是不同的。陽虛組病更多地表現(xiàn)為糖蛋白的表達異常,伏寒組更多的是體現(xiàn)在補體系統(tǒng)的異常。伏寒組經(jīng)藥物干預(yù)后,部分與人體免疫、動脈硬化相關(guān)的蛋白可以得到調(diào)整,說明早期識別及早期干預(yù)對疾病的預(yù)防具有重大意義。
[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.
【學(xué)位授予單位】：長春中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R228

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