| 【Background and aims】Hypertension is a systemic disease with arterial pressure of systemic circulation increases accompanied by functional or structural changes in heart, vascular, brain and kidney et al. International morbility of hypertension is about 25% to 30%. Hypertension has been a leading cause of illness and death in adults. Hypertensive heart disease is the main clinical manifestations of complications of hypertension with unclear molecular mechanisms even now. Because of transcription, translation and post translation modifications, results of early genomics research might not revealed at protein level. And with the methods of immunohistochemistry, only a few of proteins can be effectively analysed at the same time, which unable to meet the needs of the high throughput research.Differential proteomics is the most wildly used research strategies of proteomics. Though the comparative analysis of the proteins with differential expression between different situations or similar species, differential proteomics may find the regulations of metabolic within the body and reveal the natural disciplines of the body responding to internal and external environment changes. Differential proteomics researchs have wide application prospects in the life science community with the traditional but mature technologies of two-dimensional electrophoresis and mass spectrometry. Current studies, however, are always researching with animals like spontaneously hypertensive heart rats, and only a few of proteomic studies focusing on hypertension and hypertensive heart disease uses myocardium from human hearts.A large numbers of corpses involved in forensic pathology cases. The decedents are always been preserved under-20 ℃ environment and tissues and organs are in good conditions at autopsy and low degree of degradation of bio-macromoleculars like proteins. So those corpses provides a large number of experiment meterials for life science research. In forensic pathology cases, death caused by hypertensive heart disease is not uncommon and those hearts povide reliable human myocardium specimens for the research of hypertensive heart disease. 【Objectives】In this study, we collected the left ventricular myocardium specimens of human heart in the practice of forensic pathology, screened on the protein in myocardial tissues comprehensively with the methods of proteomics, found the differential expression of proteins associated with hypertension and hepertensive heart disease, established the research foundation of the differential expression of proteins in human heart specimens, selected some differential proteins for deep research to discuss the role of them in the formation process of hypertension and hypertensive heart disease. All of this aim to provide new clues for further research in hypertension and hypertensive heart disease.1. to establish the protein expression map of human left ventricular myocardium tissue.2. to analyze the differential expression of myocardial proteins in non-hypertension, hypertension and hypertensive heart disease conditions.3. select some proteins for mass spectrometry identification and conduct deep research to the identified proteins, discuss the role of them in the formation process of hypertension and hypertensive heart disease. 【Materials and Methods】1. collect left ventricular myocardium specimens in well preserved human hearts, devide the specimens according to the results of forensic pathology examination and the research purpose into three groups( non-hypertension, hypertension and hypertensive heart disease).2. 100 mg of left ventricular myocardium tissue from each specimens, extract cardiac total protein respectively and run two-dimensional electrophoresis and mass spectrometry for identification, and analyze the differential expression of myocardial proteins in nonhypertension, hypertension and hypertensive heart disease conditions.3. Verify the differential expression of proteins by western blotting and real-time PCR. 【Results】1. Contrast analysis of gel images between three groups revealed that there are 67 proteins up-regulate in HT3-HHD2 contrast group and 39 down-regulate, 50 proteins upregulate in NHT1-HHD2 contrast group and 32 down-regulate, and 98 proteins up-regulate in NHT1-HT3 contrast group and 103 down-regulate.2. By mass spectrometry identification, 8 groups of protein were identified, including heat shock protein B1(Hsp B1), peroxiredoxin-2(Prx2), ATP synthase subunit alpha,(mitochondrial, ATP5A1), Fibrinogen beta chain,(FIBB), alpha-1-antitrypsin,(A1AT), Vatamin D-binding protein,(VTDB), serine protease inhibitor,(Serpin B6), Voltagedependent anion-selective channel protein 1,(VDAC1).3. The verity results of western blotting and real-time PCR to Hsp B1, Prx2 and ATP5A1 were consistent with the two-dimensional electrophoresis results.4. The expression of Hsp B1 in HT group was up-regulating obviously than NHTgroup, and down-regulating obviously in HHD group than HT group, but no obvious difference between NHT and HHD groups. Prx2 expression was up-regulating obviously in HT group and then down-regulating obviously in HHD group, the expression of Prx2 in HHD group was down-regulating obviously then in NHT and HHD group; ATP5A1 expression, however, was down-regulating obviously in HT and HHD group than NHT group but no obvious difference between HT and HHD groups.【Conclusions】1. By separating the proteins in left ventricular myocardium of human heart, we obtained the protein expression map of human left ventricular myocardium in condition of non-hypertension(NHT), hypertension(HT) and hypertensive heart disease(HHD).2. Contrast analysis of gel images between three groups revealed that there are 67 proteins up-regulate in HT3-HHD2 contrast group and 39 down-regulate, 50 proteins upregulate in NHT1-HHD2 contrast group and 32 down-regulate, and 98 proteins up-regulate in NHT1-HT3 contrast group and 103 down-regulate. We identified 8 groups of protein by mass spectrometry identification.3. The difference expression of proteins between NHT, HT and HHD groups involved in exidative stress and energy metabolism.4. The identified proteins found in this research provide new clues for futher research to hypertension and hypertensive heart disease. |
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