生长分化因子15（GDF15）作为线粒体病筛选诊断标记物的研究

发布时间：2018-02-15 00:55

本文关键词： 线粒体病 GDF15 生物标记物　出处：《山东大学》2016年硕士论文　论文类型：学位论文

【摘要】：背景线粒体病是由于线粒体DNA (mtDNA)或核DNA突变导致的原发性线粒体呼吸链氧化磷酸化障碍的一组遗传代谢性疾病,在人群中患病率约为1：5000。该病在各个年龄段均可发病,而且临床表现各异。所有器官均可受累,临床上以能量需求旺盛的脑、心、肾、眼、耳及肌肉等多见。线粒体病根据临床表现可分为不同的临床综合征,临床上以线粒体脑肌病伴高乳酸血症和卒中样发作(MELAS),肌阵挛癫痫伴破碎红纤维(MERRF)和慢性进行性眼外肌麻痹(CPEO)、亚急性坏死性脑脊髓病(Leigh综合征)等多见。也可表现为多种症状的非特异性组合或者多种综合征的叠加,或者仅仅表现单一的症状,如听力下降或者糖尿病等。线粒体病非特异性的临床表现和缺乏可靠的生物标记物给临床诊断造成了很多困难。在临床工作中,我们常用的线粒体病标记物包括：乳酸,丙酮酸,肌酸激酶。然而,这些生物学标记物在诊断线粒体疾病方面效能较低,因此,目前诊断的标准仍然依靠病史和肌肉活检酶组织化学染色。肌肉病理诊断需要先进的实验室设施、熟练技术人员及大量资金的投入,因此限制了肌肉病理工作的普及。对于广大不能开展肌肉活检酶组织化学染色的医院,通过结合患者病史和可靠性的血清生物学标志物来提示我们进行基因诊断或肌肉病理诊断是十分必要的。生长分化因子15(Growth Differentiation Factor 15),也称为巨噬细胞抑制因子,是转化生长因子-β超家族成员之一,最早在活化的巨噬细胞中发现其mRNA表达升高。人体许多组织均能分泌GDF15,例如：在炎症、氧化应激,低氧等情况下心肌组织,肝组织的GDF15表达量明显升高。近几年国外有研究表明：G DF15可以作为心脏病、肝病、肿瘤等患者的非特异性血清生物学标记物。有趣的是：最近Kalko和Saskia Koene等指出血清GDF15与线粒体功能损害相关,可能是诊断线粒体病的潜在生物学标记物且与疾病临床严重程度相关。目的：本研究旨在探讨中国人群血清生长分化因子(GDF15)在线粒体病诊断筛查中的价值,同时探究血清GDF15水平与Newcastle线粒体病评分量表(Newcastle Mitochondrial Disease Adult Scale, NMDAS)的相关关系。方法：线粒体病患者42例,其平均年龄28.8±14岁。我们根据线粒体病患者年龄和性别选取了我院健康体检中心健康人与确诊的非线粒体相关的神经肌肉病作为正常对照组和对照组作为对照。20例非线粒体性神经肌肉病患者,其平均年龄39+18.9岁。正常对照组50例,平均年龄29.5±12.4岁。选取部分线粒体病患者的肌肉标本作为线粒体病组,确诊为非神经肌肉患者的肌肉标本作为正常对照组。两组之间性别、年龄比较差异无统计学意义(P0.05)。对所有入选者均抽取空腹外周静脉血5ml,3000g/min离心10分钟,留取1ml血清标本分装、编码后于-80℃冰箱保存用于GDF15测定。比较每个生物标记物水平(GDF15, FGF21、乳酸、肌酸激酶)在不同组间的差异；通过诊断性比值比(OR)、敏感性、特异性、阳性预测值、阴性预测值、诊断性OR、ROC曲线等比较GDF、FGF21、乳酸、肌酸激酶在线粒体病诊断方面的价值。结果：1.线粒体病患者血清GDF15水平均明显高于病例对照组和正常组。2.血清GDF15的诊断比值比(OR)明显高于FGF21,乳酸和肌酸激酶。3.线粒体病患者肌肉中GDF15的mRNA表达量明显升高。4.血清GDF15水平与线粒体病患者肌肉病理中破碎红纤维的比例呈正相关。5.血清GDF15水平与线粒体病患者临床严重程度(NMDAS评分)呈正相关。6.ROC曲线下的面积(AUC):GDF15:AUC 0.998 [95% CI 0.993-1], p0.001), FGF21:AUC 0.925 ([95%CI 0.860-0.966], p0.001),乳酸：AUC 0.870 (95% CI [0.794-0.926], P＜0.001),肌酸激酶：AUC 0.553 (95% CI [0.452-0.650], p= 0.3774).结论：血清GDF15是线粒体病的敏感标记物,可以作为首要检查方法来辅助诊断线粒体病。
[Abstract]:The background of mitochondrial disease is due to mitochondrial DNA (mtDNA) is a group of inherited metabolic diseases or nuclear DNA mutations in primary oxidative phosphorylation of the mitochondrial respiratory chain disorders, prevalence rate is about 1:5000. of the disease in all ages can occur in the crowd, but with different clinical manifestations. All the organs can be involved in clinical, energy the demand for the brain, heart, kidney, eye, ear and muscle rare. Mitochondrial disease according to the clinical manifestations can be divided into different clinical syndrome, clinically with mitochondrial encephalomyopathy with lactic acidosis and stroke like episodes (MELAS), muscle clonic epilepsy with ragged red fibers (MERRF) and chronic progressive extraocular muscle paralysis (CPEO), subacute necrotizing encephalomyelopathy (Leigh syndrome) and other rare. Also could be expressed in a variety of symptoms or a combination of a variety of non-specific syndrome superimposed, or just single symptoms, such as hearing loss or Diabetes. Mitochondrial disease nonspecific clinical manifestations and lack of reliable biomarkers has caused many difficulties for clinical diagnosis. In clinical work, we used mitochondrial disease markers including: lactate, pyruvate, creatine kinase. However, these biological markers of efficacy in diagnosis of mitochondrial disease is low, therefore, at present, the standard of diagnosis still depends on the history and muscle biopsy histochemical staining. The muscle pathological diagnosis need advanced laboratory facilities, skilled technical personnel and a large number of funds, thus limiting the popularity of muscle pathological work. For the majority of the hospital can not carry out muscle biopsy histochemical staining, by combining with the history and reliability of biochemiccal the marker to prompt us to perform gene diagnosis or pathological diagnosis is very necessary. The growth differentiation factor 15 (Growth Differe Ntiation Factor 15), also known as macrophage inhibition factor, transforming growth factor beta superfamily, most found that the increased expression of mRNA in activated macrophages. Many human tissues can secrete GDF15, for example: in inflammation, oxidative stress, myocardial tissue hypoxia condition, the expression of GDF15 in liver tissue volume increased obviously. In recent years, foreign studies have shown that: G DF15 can be used as heart disease, liver disease, nonspecific serum biomarkers for tumor patients. It is interesting: recently Kalko and Saskia Koene indicated that the correlation between serum GDF15 and mitochondrial dysfunction may be potential biomarkers for diagnosis of mitochondrial diseases and with the severity of clinical disease. Objective: This study aimed to investigate Chinese serum growth differentiation factor (GDF15) in the diagnosis of mitochondrial disease screening. At the same time, to explore the level of serum GDF15 and N Ewcastle mitochondrial disease rating scale (Newcastle Mitochondrial Disease Adult Scale, NMDAS) of the relationship. Methods: 42 cases of patients with mitochondrial disease, the average age of 28.8 + 14 years. According to the age and sex of patients with mitochondrial disease from healthy people and diagnosed in our hospital health examination center of non mitochondrial related neuromuscular disease as normal the control group and the control group as the control cases of non.20 patients with mitochondrial neuromuscular disease, the average age of 39+18.9 years. 50 cases of normal control group, mean age 29.5 + 12.4 years. Patients were part of mitochondrial diseases as the underlying muscle mitochondrial disease were diagnosed as non neuromuscular patients with muscle specimens as the control group. Sex between the two groups, no significant age difference (P0.05). All the subjects were fasting peripheral venous blood 5ml, 3000g/min centrifuge for 10 minutes, take the 1ml standard serum The packaging and encoding in -80 DEG C refrigerator for determination of GDF15. The comparison of each biomarker levels (GDF15, FGF21, creatine kinase, lactic acid) differences in different groups; through the diagnostic odds ratio (OR), sensitivity, specificity, positive predictive value, negative predictive value, diagnosis of OR, ROC curve comparison of GDF, FGF21, lactate, creatine kinase value in the diagnosis of mitochondrial diseases. Results: 1. patients with mitochondrial disease serum GDF15 levels were significantly higher than the diagnostic ratio of case control group and normal group.2. serum GDF15 ratio (OR) was significantly higher than that of FGF21, GDF15 lactic acid and creatine kinase.3. in muscle of patients with mitochondrial disease mRNA the expression of.4. significantly increased serum GDF15 level and mitochondrial disease in patients with muscle pathology in ragged red fiber ratio of.5. was positively correlated with serum GDF15 level in patients with mitochondrial disease severity (NMDAS score) was positively related to.6.ROC under the curve Area (AUC): GDF15:AUC 0.998 [95% CI 0.993-1], p0.001 FGF21:AUC 0.925 ([95%CI), 0.860-0.966], p0.001, AUC (0.870): lactic acid 95% CI [0.794-0.926], P < 0.001), creatine kinase ([0.452-0.650]: AUC 0.553 CI 95%, p= 0.3774). Conclusion: serum GDF15 is a sensitive marker of mitochondrial diseases, can as the primary examination method to diagnosis of mitochondrial diseases.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R746

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