2型糖尿病并发非酒精性脂肪性肝炎患者血清脂质组学分析及诊断价值研究

Serum Lipomics Analysis and Diagnostic Value of Type 2 Diabetes Patients with Nonalcoholic Steatohepatitis

目的 探讨2型糖尿病(T2DM)并发非酒精性脂肪性肝炎(nonalcoholic steatohepatitis, NASH)患者血清脂质组学分析及诊断价值。方法 选取2018年4月~2020年4月中国人民解放军联勤保障部队第九二一医院收治的80例T2DM并发非酒精性脂肪性肝病(nonalcoholic fatty liver disease, NAFLD)患者,根据肝活检结果分组,其中T2DM并发NASH患者38例(NASH组),并发非酒精性单纯性脂肪肝(NAFL)患者42例(NAFL组)。采用超高效液相色谱-串联质谱法(UPLC-MS/MS)进行血清脂质组学分析,并比较其检测结果,同时采用Logistic逐步回归分析及受试者工作特征(ROC)曲线对T2DM发生NASH的潜在血清标志物进行筛选。结果 与NAFL组比较,NASH组碱性磷酸酶(ALP)(98.79±29.65 U/L vs 82.94±18.66U/L)、天冬氨酸氨基转移酶(AST)(32.12±13.64 U/L vs 21.33±15.51U/L)、γ-谷氨酰基转肽酶(GGT)(86.87±24.38 U/L vs 45.06±15.22U/L)、晨起空腹血糖(FPG)(10.20±3.15mmol/L vs 7.72±1.79mmol/L)明显升高,差异具有统计学意义(t=4.176~33.168,均P<0.05);脂质学分析NASH组磷脂酰胆碱(PC)(38∶3),PC(35∶4),PC(36∶1)^(a),PC(36∶1)^(b),PC(44∶5)、磷脂酰乙醇胺(PE)(34∶1)^(c),PE(34∶1)^(d),PE(36∶2),PE(38∶1),PE(39∶1),磷脂酰丝氨酸(PS)(33∶0)、三酰甘油(TG)(52∶1)、TG(54∶2)、鞘磷脂(SM)(37∶1),SM(39∶1),SM(40∶1)、葡萄糖神经酰胺(GlcCer)(40∶2)和棕榈酰苯丙氨酸水平均明显高于NAFL组(VIP=1.275~3.176,均P <0.05);NASH组PC(38∶5),PC(38∶6),TG(52∶4),TG(54∶4),TG(54∶5),TG(57∶6),TG(58∶4)和TG(60∶6)水平明显低于NAFL组(VIP=1.086~3.086,均P <0.05);Logistic逐步回归分析显示:PC(36∶1)a和PE(38∶1)升高是T2DM患者进展为NASH的危险因素(P<0.05);PC(36∶1)a和PE(38∶1)ROC曲线下面积分别为0.814,0.792,敏感度分别为94.28%,94.86%,特异度分别为77.65%,76.31%,均高于GGT,AST和ALT。结论 T2DM并发NASH患者,出现变化的主要脂质是甘油磷脂、鞘脂和TG,其中PC(36∶1),PE(38∶1)可能成为诊断NASH的潜在血清标志物。

Objective To investigate the value of serum lipoomics analysis and diagnosis in type 2 diabetes mellitus(T2DM)patients with nonalcoholic steatohepatitis(NASH).Methods 80 patients with T2DM combined with nonalcoholic fatty liver disease(NAFLD)admitted to the 921st Hospital,Joint Logistics Support Force of the Chinese People’s Liberation Army from April 2018 to April 2020,and grouped on the liver biopsy results.Among them,38 patients with T2DM combined with NASH(NASH group),and 42 patients with non-alcoholic simple fatty liver disease(NAFL)(NAFL group).The serum lipids were analyzed by ultra-high performance liquid chromatography-mass spectrometry and the detection results were compared.At the same time,the potential serum markers of NASH in T2DM were screened by logistic stepwise regression analysis and subject working characteristics(ROC).Results Compared with NAFL group,in NASH group,ALP(98.79±29.65 U/L vs 82.94±18.66 U/L),AST(32.12±13.64 U/L vs 21.33±15.51 U/L),γ-GGT(86.87±24.38 U/L vs 45.06±15.22U/L)and FPG(10.20±3.15 mmol/L vs 7.72±1.79 mmol/L)increased significantly in the morning,with significant difference(t=4.176~33.168,all P<0.05).Lipographic analysis of NASH group phosphatidylcholine(PC)(38∶3),PC(35∶4),PC(36∶1)^(a),PC(36∶1)^(b),PC(44∶5),phosphatidylethanolamine(PE)(34∶1)^(c),PE(34∶1)^(d),PE(36∶2),PE(38∶1),PE(39∶1),phosphatidylserine(PS)(33∶0),triacylglycerol(TG)(52∶1),TG(54∶2),sphingomyelin(SM)(37∶1),SM(39∶1),SM(40∶1).The levels of glucose ceramide(GlcCer)(40∶2)and palmitoyl phenylalanine were significantly higher than those in NAFL group(VIP=1.275~3.176,all P<0.05).The levels of PC(38∶5),PC(38∶6),TG(52∶4),TG(54∶4),TG(54∶5),TG(57∶6),TG(58∶4)and TG(60∶6)in NASH group were significantly lower than those in NAFL group(VIP=1.086~3.086,all P<0.05).Logistic stepwise regression analysis showed that the increase of PC(36∶1)a and PE(38∶1)was a risk factor for T2DM patients to develop NASH(P<0.05).The area under ROC curve of PC(36∶1)a and PE(38∶1)were 0.814 and 0.792,the sensitivity were 94.28%and 94.86%,and the specificity were 77.65%and 76.31%,respectively,higher than GGT,AST,ALT.Conclusion In T2DM patients with NASH,the main lipid changes were glycerophospholipid,sphingolipid and TG.PC(36∶1)and PE(38∶1)may become potential serum markers for the diagnosis of NASH.