应用液相色谱-串联质谱法系统检测糖尿病肾病和糖尿病神经病变患者尿儿茶酚胺水平

Systematic assessment of urinary catecholamines in type 2 diabetic nephropathy and neuropathy patients by liquid chromatography tandem mass spectrometry

目的研究尿儿茶酚胺水平与Ⅱ型糖尿病肾病及神经病变的关系.方法病例对照研究.收集华中科技大学同济医学院附属同济医院收治的Ⅱ型糖尿病患者130例(年龄27~81岁,男75例,女55例)和健康对照118名(年龄27~76岁,男67名,女51名)的24h尿液.另收集Ⅱ型糖尿病患者115例(年龄27~77岁,男75例,女40例)和健康对照93名(年龄25~74岁,男57名,女36名)的随机尿.根据尿白蛋白清除率的检测结果,Ⅱ型糖尿病患者分为肾病组(24h尿:37例,随机尿:32例)和非肾病组(24h尿:32例,随机尿:35例);根据问卷调查,定量感觉试验及自主功能试验,Ⅱ型糖尿病患者分为神经病变组(24h尿:31例,随机尿:23例)和非神经病变组(24h尿:30例,随机尿:25例).采用液相色谱-串联质谱法(LC-MS/MS)测定尿儿茶酚胺(CAs)的含量,包括肾上腺素(E)、去甲肾上腺素(NE)、多巴胺(DA)和总儿茶酚胺(总CAs)的含量,并探讨不同分组患者儿茶酚胺的水平.采用χ2检验、独立样本t检验和Mann-Whitney非参数检验对结果进行分析.结果Ⅱ型糖尿病肾病患者24h尿液E(0.74±0.24)μg/24h,NE(9.22±4.02)μg/24h,DA(64.77±21.68)μg/24h和总CAs(74.72±25.65)μg/24h低于非肾病组[E:(4.23±0.50)μg/24h,U=10,P<0.001;NE:(32.31±1.74)μg/24h,t=-2.72,P=0.011;DA:(219.58±27.51)μg/24h,t=-2.88,P=0.007;总CAs:(256.02±30.65)μg/24h,t=-3.02,P=0.005]且差异具有统计学意义(P<0.05).在随机尿液中糖尿病肾病组NE、DA和总CAs含量与非肾病组的差异亦具有统计学意义(P<0.05).结论本研究结果表明糖尿病肾病患者尿儿茶酚胺含量低于非肾病患者,可能与糖尿病肾病的发生发展有关.

Objective To investigatethe relationship between urinary catecholamine levels and type Ⅱ diabetic nephropathy and neuropathy. Methods Allsubjects were collected fromTongji Hospital, Tongji Medical College, Huazhong University of Science and Technology,and were divided into two groups: type Ⅱ diabetic patients (For 24 h urine, n=130, aged 27-81 years old, 75 men, 55 female;for random urine, n=115, aged 27-77 years old, 75 men, 40 female) and healthy control (For 24 h urine, n=118, aged 27-76 years old, 67 men, 51 female;for random urine, n=93, aged 25-74 years old, 57 men, 36 female).Based on the results of urinary albumin excretion, type Ⅱ diabetic patients were prospectively divided into two independent groups: patients with nephropathy (For 24 h urine, n=37;for random urine, n=32) and patients without nephropathy (For 24 h urine, n=32;for random urine, n=35). According to questionnaires, quantitative sensory testing and autonomic function test, type Ⅱ diabetic patients were divided into two different groups: patients with neuropathy (For 24 h urine, n=31;for random urine, n=23) and patients without neuropathy (For 24 h urine, n=30;for random urine, n=35). Urinary catecholamines(CAs) levels, including epinephrine (E), norepinephrine (NE), dopamine (DA) and total catecholamines (total CAs) levels, were measured using liquid chromatography tandem mass spectrometry (LC-MS/MS). The CAs levels in different groups have been discussed. The results were analyzed using χ2 test, independent t test and Mann-Whitney non-parametric test. Results Type Ⅱ diabetic patients with nephropathy show lower E (0.74±0.24)μg/24 h, NE(9.22±4.02)μg/24 h, DA(64.77±21.68)μg/24 h and total CAs(74.72±25.65)μg/24 h in 24-hour urine in comparison with the group without nephropathy(For E, 4.23±0.50 μg/24 h, U=10, P<0.001;for NE,(32.31±1.74)μg/24 h, t=-2.72, P=0.011;for DA,(219.58±27.51)μg/24 h, t=-2.88, P=0.007;fro CAs,(256.02±30.65)μg/24 h, t=-3.02, P=0.005) and the differences were statistically significant (P<0.05). The differences of NE, DA and total CAs between type Ⅱ diabetic patients with nephropathyand without nephropathy were statistically significant in random urine(P<0.05). Conclusion These results suggested that urine CAs levels in diabetic patients with nephropathy is lower than that in patients without nephropathy, which may be related to the occurrence and development of diabetic nephropathy.