10 血浆蛋白质以及非蛋白含氮化合物的代谢紊乱

nullThe Plasma Proteins (血浆蛋白)The Plasma Proteins (血浆蛋白) M.E. Mothobi Clinical Laboratory, Zhongnan Hospital 2008.4.14 Contents Contents1. Concept of Plasma Proteins (血浆蛋白的概念) 2. Total Protein (总蛋白) Acute-phase Reactants (急性期蛋白) Individual Proteins and Disease States (个别蛋白质和疾病状态) Methods of Protein Analysis (蛋白方法)1.Concept of plasma proteins1.Concept of plasma proteinsThe plasma proteins are a diverse group of molecules that perform a variety of functions. 血浆蛋白是一组不同的分子,并执行许多种功能 Grouped as related molecules that collectively carry out a specific function. 相关分子成为一组,执行特殊的功能 Need to be distinguished from proteins that occur only incidentally in the blood.需要区别于偶然出现于血中的蛋白质Classification of plasma proteins (血浆蛋白分类)Classification of plasma proteins (血浆蛋白分类)Often Classified Based on Their Electrophoretic Mobility On Cellulose Acetate Or Agarose Gel. 根据它们在乙酸纤维素上或琼胶素明胶电泳来分类 Five Regions (albumin,α1,α2,β and γ regions). 五个区 (清蛋白, α1,α2,β和γ区) Most of Them are Synthesized in the liver 大部分在肝脏合成 Normal Serum Protein ElectrophoresisNormal Serum Protein ElectrophoresisSelected major and minor plasma proteinsSelected major and minor plasma proteinsProtein Abnormalities(蛋白质异常)Protein Abnormalities(蛋白质异常)Plasma protein abnormalities arise from one or more of the following: (在以下情况中血浆蛋白会出现异常) Congenital(先天性)abnormalities affecting a specific protein ( e.g., AAT deficiency) 先天异常导致产生特殊基因 Acquired (获得的)abnormalities affecting a specific protein ( e.g., transferrin in anemia) Alterations affecting multiple proteins reflecting variations in the physiologic state e.g., age, gender, pregnancy (生理状态下,还有其他因素会导致多种蛋白的变异) Alterations affecting multiple proteins secondary to disease (e.g., chronic liver disease) (也可能继发与其他疾病) 2. Total Protein(TP)2. Total Protein(TP)The serum total protein includes both albumin (60%) and globulin (40%).血清总蛋白包括60%白蛋白和40%球蛋白 TP concentration can be measured on either serum or plasma (serum is preferred).血清总蛋白浓度可以用血清或血浆来测定,但一般先选用血清. Factors Which Affect Protein Concentration: 影响蛋白浓度大因素: * Fluid Balance(体液平衡). * Changes in Synthesis or Catabolism(合成或分解代谢的改变) * Protein losses(蛋白质的丢失). Albumin : Globulin ratio is more preferred.测量白/球比值比测量总蛋白要好.Pre-analytical errors which may alter the serum protein value Pre-analytical errors which may alter the serum protein value Prolonged tourniquet application during venipuncture increases the total protein by as much as 5g/L due to hemoconcentration.静脉穿刺时,由于长时间应用止血带会使血液浓缩,从而导致总蛋白升高5 g/L. Recumbent subjects may be 10% lower than in ambulatory individuals.取血时,卧位的病人其蛋白总数比能活动的病人要低10%. Normal diurnal variation of 4 g/L and a seasonal fluctuation: Peak in Nov. and Lowest in June. 有日夜变化大约4 g/L 和季节变化,11月最高,6月最低. Exercise increases the serum protein concentration by as much as 10%, but this effect is transient. 运动会使血清蛋白浓度增高约定10%, 但很快又恢复正常. nullHypoproteinemia(低蛋白血症) * decreased protein synthesis(蛋白质合成减少) * malnutrition(营养不良) * chronic liver disease(慢性肝病) Nephritic syndrome(肾炎综合征) * the serum albumin level may fall to less than 5 g/L 血清白蛋白水平会下降到5 g/L 以下 Acute Phase Reactants (APRs) (急性期蛋白)Acute Phase Reactants (APRs) (急性期蛋白)APRs are a group of proteins whose plasma concentration changes in response to a variety of inflammatory states including: 急性期蛋白是包括对多种炎性状态下其血浆浓度发生变化的一组蛋白质。 * Infection(感染) * Surgery(手术) * Trauma(创伤) * Myocardial Infarction(心肌梗死) * Malignancy(恶性肿瘤) * Any Condition Associated with Tissue Necrosis（任何时候都伴随着有组织坏死）Regulation (调节) of APRsRegulation (调节) of APRsCytokines, major stimulator is IL-6. 细胞因子, 主要刺激因子是IL-6 Other cytokines: 其他细胞因子: *Interleukin-1β (IL-1β)（白细胞介素1β） *Tumor Necrosis Factor-α (TNF-α)（肿瘤坏死因子α） *Interferon Gamma (IFN-γ) （干扰素γ） *Transforming growth factor-β (TGF-β)(转化生长因子) Up-regulate the subgroups of APRs. 上述因子可以上调APRs的亚群 C-reactive protein appears in the serum in 4-6 hours, other proteins follow in a day or so. （C反应蛋白4-6小时出现,其他蛋白一天或之后才会出现.） Major acute phase reactantsMajor acute phase reactantsMeasurement (检测) of APRs C reactive protein (CRP)Measurement (检测) of APRs C reactive protein (CRP)CRP was first identified in 1930 as a substance present in the sera of patients with pneumococcal pneumonia that could bind to C-polysaccharide isolated from Streptococcus pneumoniae, producing a flocculation reaction. CRP是1930年在肺炎球菌肺炎病人血清中以A血型红细胞的凝集原物质被鉴定出来的，它可以和肺炎链球菌中分离的C型多糖结合，产生絮凝反应）。 C-reactive protein (CRP)C-reactive protein (CRP)The early rise in CRP together with the largest factor by which it increases makes it the most useful APR to measure in the clinical lab. 实验室里最有用的APR就是CRP,因为它会较早出现,且数值会非常高 It is synthesized by the liver, short half life of 19 hrs.它在肝脏里合成，短半衰期为19小时 C- reactive protein (CRP)2C- reactive protein (CRP)2Functions: Several functions including;（功能：包括） Initiation of opsonization and phagocytosis by phagocytes吞噬细胞的调理和吞噬功能的启动 Complement activation（补体的激活） Recognition of necrotic tissue（识别坏死组织) Clinical significance: Elevated in a variety of acute inflammatory diseases.（多种急性炎性疾病中升高。） Follow up of patients with acute inflammatory diseases. 急性炎症疾病的患者可以测量CRP来观察其病情 Reference range: Less than 0.8mg/dL. 2. Individual proteins and disease states2. Individual proteins and disease statesPre-albumin(前白蛋白) Albumin(白蛋白) α1-Antitrypsin (α1-抗胰蛋白酶) α2-Macroglobulin(α2-巨球蛋白) Ceruloplasmin(铜蓝蛋白) Haptoglobin (结合珠蛋白) Transferrin(转铁蛋白) β-Lipoprotein (β -脂蛋白) β2-Microglobulin(β2-微球蛋白)Pre-albumin前白蛋白Pre-albumin前白蛋白Pre-albumin (MW 55,000) is a tryptophan-rich tetrameric non-glycosylated protein named for its anodal migration relative to albumin on protein electrophoresis. 前白蛋白是一种富含色氨酸的去糖基的四聚体，其名称的由来是其电泳的迁移相对于白蛋白而言的。 Synonyms include transthyretin and thyroid binding pre-albumin. 前白蛋白又包括血液中三种甲状腺激素结合蛋白之一和甲状腺前白蛋白. Function: carrier protein for thyroid hormone and for vitamin A (with Retinol Binding Protein). [(功能:是甲状腺激素和维生素A (视黄醇结合蛋白)的运载者)] Synthesis: in the liver with a half life of 1.9 days. (在肝脏合成,半衰期待1.9天) Pre-albumin 2Pre-albumin 2Pre-albumin is a compact molecule that can cross the blood-brain barrier and may also be synthesized by the cells of the choroids plexus. （前白蛋白是一种能穿越血脑屏障的致密分子，脉络丛的细胞也能够合成这种蛋白质)。 Pre-albumin can be measured by a variety of immunoassays, including radial immunodiffusion (RID) and nephelometry. 前白蛋白可以用多种免疫测定方法检测:放射免疫扩散和用悬液计测量法 Pre-albumin levels reflect hepatic synthesis and can serve as an index of liver function. 前蛋白水平可以反映肝合成情况,也可以作为肝功能的指标 Reference Range: 195 - 358 mg/L Albumin白蛋白Albumin白蛋白The most abundant protein in plasma, comprising approximately 60% of the total protein concentration. 它是血浆中含量最多大蛋白,占总蛋白的60% Alb consists of a single chain of 580 amino acids with a molecular mass of 66.3kD. 白蛋白包括580氨基酸,其分子量为66.3kD Synthesis: exclusively by the liver. Half life 19 days. 在肝脏合成,半衰期待19天 Albumin 2Albumin 2Functions: 1. 80% of the plasma COP. 80%胶体渗透压 2. Amino Acid Source to a Variety of Cells 多种细胞的氨基酸来源 3. Major Transport Protein 主要的运输蛋白,e.g., 胆红素 4. Lipid Metabolism 参与脂类代谢 Reference Range: 31 to 43g/L. Albumin 3Albumin 3Measurement: Electrophoretic Immunochemical dye-binding techniques bromocresol purple Bromocresol greenAlbumin 4Albumin 4Clinical significance: 临床意义: Decreased levels may result from a variety of causes: 很多因素可以导致白蛋白水平下降: *liver disease (impaired synthesis). [(肝病(合成受损)] *excessive loss (nephritic syndrome).丢失过多(肾炎综合征 *hemorrhage, burns. (出血, 烧伤) *exudative processes, especially of the skin and GIT. 渗出性病变, 尤其是皮肤和胃肠道 *third spacing (ascites, effusions). (腹水, 渗出液)α-1 Antitrypsin (AAT) α1-抗胰蛋白酶α-1 Antitrypsin (AAT) α1-抗胰蛋白酶AAT is one of the major plasma proteins, comprising nearly 90% of the α1 globulin region on SPE.(AAT是一种主要的血浆蛋白，由SPE上的近90%的α1球蛋白组成)。 Belongs to a family of serum proteins collectively known as serpins (serine protease inhibitor 丝氨酸蛋白酶抑制剂). 属于血清蛋白家族，共同的以丝氨酸蛋白酶抑制剂被认识。Genes encoding the AAT protein comprise an autosomal allelic system containing at least 75 codominant genes inherited on a single locus called Pi for proteinase inhibitor. AAT蛋白的基因编码包括一个常染色体等位系统，它包含了至少75个由一个叫Pi的蛋白酶抑制因子单基因转坐遗传而来的共显性基因。 α-1 Antitrypsin (AAT) 2α-1 Antitrypsin (AAT) 2The PiZ allele is the most common variant associated with AAT deficiency. PiZ等位基因随着AAT的变化而变化是很常见的。 Homozygous PiZZ : 10% to 15% of normal 正常的纯合子PIZZ：10%-15% Heterozygotes PiMZ: 60% of normal 正常的杂合子PiMZ：60% Rhe phenotypes PiSS and PiMS have levels of 63% and 83% of normal respectively.（Rhe表型的Piss和PiMS各自分别有63%和83%的正常水平）α-1 Antitrypsin (AAT) 3α-1 Antitrypsin (AAT) 3Synthesis: In the Liver. It can also be found in Tears, Lymph, Bile, Semen, and Amniotic.（合成：在肝脏。同时在眼泪，淋巴液，胆汁，精液和羊水里也能找到）。 Function: Proteinase inhibitor mainly non trypsin proteases e.g., elastases.（功能：蛋白酶抑制因子主要是非胰蛋白酶朊酶，例如弹性蛋白酶）。 Reference Range: 900~1500mg/L（参考范围） Deficiency may be congenital or acquired.（不足可能是先天性的或者是获得性的）。 Congenital deficiency is associated with pulmonary emphysema and hepatic cirrhosis.（先天性的不足常伴随着由肺气肿和肝硬化）。 α-1 Antitrypsin (AAT) 4α-1 Antitrypsin (AAT) 4Acquired decreases are seen in:（获得性的不足常导致） Protein-losing syndromes, (蛋白质丢失综合症) malnutrition, （营养不良） severe liver damage, （严重的肝细胞损伤） respiratory diseases: neonatal respiratory distress syndrome（呼吸系统疾病：新生儿呼吸窘迫综合症） α-2 Macroglobulin (AMG) α2-巨球蛋白 α-2 Macroglobulin (AMG) α2-巨球蛋白AMG Consists of four identical subunits and is one of the largest serum proteins (MW 720,000).（AMG包括四个相同的亚基，它是最大的血清蛋白之一）。 Synthesis: In the liver. Half life of 5 days.（合成：在肝脏，半衰期5天）。 Function: * Defense Against Proteolytic Enzymes (Irreversibly Binds Proteinases )（抵抗蛋白水解酶） * Can inhibit Several endopeptidases （能抑制若干蛋白水解酶） * Can bind numerous growth factors, hormones, and cytokines (e.g., TGF- 1&2, PDGF, TNF-αe.t.c)（能和多种生长因 子，激 素，细胞活素相结合 ） Reference Range: 125 to 215 mg/dL（参考值范围） Levels are greatly increased in nephritic syndrome, may be >1000mg/dL.L Ceruloplasmin (CER)铜蓝蛋白 Ceruloplasmin (CER)铜蓝蛋白CER (MW 132,000) is a glycoprotein synthesized by the liver as a single polypeptide chain to which six to eight copper atoms are attached. （CER是由肝脏利用一种连接有6到8个铜原子的多肽链合成的一种糖蛋白）。 The pure protein is blue because of its high copper content. Increased levels may impart a green tinge to plasma samples.（这种纯蛋白是蓝颜色的，因为它由很高的铜原子含量。增加水平可以使血浆样品呈绿颜色）。 Function: *Oxidation of Fe2+ to Fe3+ (功能：氧化Fe2+ 到 Fe3+） * Prevention of lipid oxidation（预防脂质氧化） Ceruloplasmin ( CER) 2Ceruloplasmin ( CER) 2Clinical Significance: （临床意义） * Diagnosis of Wilson’s Disease （诊断肝豆核变性病） (Hepatolenticular Degeneration) Reference Range: 27 to 50 mg/dL（参考值范围） Highest in young children and somewhat lower in both Infants and Adults. （在幼儿体内最高，在婴儿和成人体内低些）。 Values less than 10 mg/dL are Suggestive of Wilson’s Disease.（小于10 mg/dL提示肝豆核变性病）。Haptoglobin结合珠蛋白Haptoglobin结合珠蛋白A glycoprotein synthesized by the liver that migrates as an α2 globulin. （一种在肝内合成的由α2球蛋白迁移而来的糖蛋白）。 In addition to its well-known function in the binding of free hemoglobin, this APR might also play an important role in the control of local inflammation. (除此之外，它以能和游离血红蛋白结合的功能被认识，这种APR可能在控制局部感染过程中扮演着重要的角色)。 haptoglobin and hemopexin protect the body against reactive oxygen species. [结合珠蛋白和血红素蛋白, 它们能对抗体内的活性氧物质(e.g., H202, singlet oxygen, free radicals, superoxides)] 。 Haptoglobin 2Haptoglobin 2Clinical Significance:（临床意义） Decreased levels are associated with intravascular hemolysis including hemolytic anemias, hemoglobin-opathies, hemolytic transfusion reactions, extensive burns, malaria, disseminated intravascular coagulation, and exercise-induced hemolysis. 水平降低常见于：血管内溶血包括溶血性贫血，血红蛋白病，溶血性输血反应 ，广泛的烧伤，疟疾，弥散性血管内凝血，运动诱导性溶血 Reference range: 16 to 199 mg/dL.（参考值范围） Transferrin转铁蛋白Transferrin转铁蛋白A glycoprotein (MW 79,550) containing two iron binding sites on a single polypeptide chain.（是一种在单多肽链上包含有2个铁结合位点的糖蛋白）。 Transport of iron from intestine (or Hemoglobin catabolism) to red cell precursors.（将铁从肠运输到红细胞前体的运输工具）。 It is synthesized mainly by the liver with half life of approx. 8 days.（主要在肝里面合成，半衰期约8天）。 Evaluation of anemia（贫血的评估） Elevated levels in Iron deficiency anemia（缺铁性贫血的提高水平） Reference Range:1.91 to 3.65g/L.（参考值范围）-Lipoprotein β -脂蛋白-Lipoprotein β -脂蛋白Lipoproteins constitute a family of molecules composed of lipids and proteins whose function is to transport cholesterol, triglycerides, and phospholipids in the blood.（脂蛋白是一种由有着运输血液里的胆固醇，甘油三酯，磷脂功能的脂质和蛋白质分子家族组成的）。 Four categories: Chylomicrons, VLDL, LDL and HDL.（分为4类：乳糜微粒，极低密度脂蛋白，低密度脂蛋白，高密度脂蛋白）。 2- Microglobulin β2-微球蛋白2- Microglobulin β2-微球蛋白A small protein (MW 11,818) comprising the common light chain of the class I major histocompatibility complex antigen on the surfaces of all nucleated cells. （是由有着共同有核细胞核上的I类MHC复合抗原的轻链组成的小蛋白质）。 Necessary for insertion of the HLA into cell membrane, and stabilizes the HLA heavy chain. It may also play a role in the regulation of some lymphocyte function.（是HLA插入细胞膜所必须的，还可以稳定HLA重链。它在调节一些淋巴细胞功能上起着重要的作用）。 Reference Range: [Serum 2.8mg/L, urine 0.2mg/L] 2-Microglobulin22-Microglobulin2Clinical significance: Elevated levels of urinary 2M are seen in patients with tubulointerstial disorders e.g., heavy metal toxicity, upper urinary tract infection, renal allograft rejection and anticancer drug toxicity. 临床的:小管间质性病症 e.g., 重金属毒性上泌尿道感染肾脏的同种异体移植物排斥抗癌药的提高水平尿的2M Elevated plasma 2M levels are seen in renal failure based on glomerular disfunction, patients with leukemia, lymphoma and some solid tumors. 肾衰竭,血管小球的机能失常,白血病,淋巴瘤和实体瘤的提高水平血浆2M 3. Methods of protein analysis3. Methods of protein analysisMeasurement of Total Protein Serum Protein Electrophoresis Other methodsMeasurement of total proteinMeasurement of total proteinbiuret method (widely used procedure) dye-binding procedures turbidimetric method nephelometric method Biuret methodBiuret methodPrinciple based on the reaction of peptide bonds with copper ions (cu 2+) in alkaline solution forming a pink to violet complex that is measured spectrophotometrically at 540nm. Intensity of the color is proportional to the protein concentration. Lacks sensitivity for use on low concentration samples such as CSF and Urine. Serum protein electrophoresisSerum protein electrophoresisPrinciples of Electrophoresis Agarose Gel Electrophoresis Capillary Electrophoresis Principles of electrophoresisPrinciples of electrophoresisSeparates molecules based on their mobility in an electric field. Support media in use include agarose gel, paper Mobile phase is composed of buffer ions Negatively charged molecules migrate towards the anode Direction of protein migration depends on the PH of the buffer and the isoelectric point of the proteinsPrinciples of gel electrophoresisPrinciples of gel electrophoresisAgarose gel electrophoresisAgarose gel electrophoresisBecause the proteins in an agarose gel are invisible, a staining solution is used to visualize the protein bands.（蛋白质在琼脂糖凝胶电泳是不可见的，所以在蛋白质带可视化的过程中用到染色剂）。 Separates proteins into five bands.（将蛋白质分离为5个条带）。Typical serum protein electrophoresis reference pattern Typical serum protein electrophoresis reference pattern Representative patterns of serum protein seen on serum protein electrophoresisRepresentative patterns of serum protein seen on serum protein electrophoresisCapillary electrophoresis （毛细管电泳）Capillary electrophoresis （毛细管电泳）Electrophoretic separation of analytes is carried out in fused-silica capillaries with an inner diameter of 50 μm (range, 20 to 200 μm), an outer diameter of 375 μm, and an effective length of approximately 50 cm (range, 7 to 100 cm). （分析物的电泳分离是在用内径为50μm，外径为375 μm，有效长度大约为50cm融合硅胶毛细管里实现的）。Other methods（其他方法）Other methods（其他方法）Ion Exchange Chromatography（离子交换层析） Affinity Chromatography（亲和层析） Isoelectric Focusing（等电聚焦）Chromatography methods（色谱法）Chromatography methods（色谱法）All forms of chromatography contain the mobile phase, which provides the driving force to separate molecules and the stationary phase which may partially or completely retards their movement thus permitting separation. 所有形式的色谱法都包括流动相，它是分离分子的驱动力;还包括固定相，它是部分的或者完全的阻止它们运动，从而使不同分子得到分离。null