摘要
目的:探究尿液细胞外囊泡中关键蛋白细胞骨架相关蛋白4(cytoskeleton⁃associated protein 4,CKAP4)作为生物标志物预测糖尿病肾病(diabetic kidney disease,DKD)进展的能力。方法:纳入南京医科大学第一附属医院143例肾活检证实DKD的2型糖尿病患者以及肾脏恶性肿瘤患者10例。免疫组化法检测肾组织中CKAP4的表达水平。采用Spearman相关分析CKAP4表达水平与临床指标的相关性。计算受试者工作特征(receiver operating characteristic,ROC)曲线下面积(area under the curve,AUC),以评估CKAP4表达是否能有效区分肾脏预后不良的患者。采用ROC曲线分析确定预测肾脏事件的CKAP4评分的最佳临界值(最高约登指数)。此外,进行时间依赖性AUC分析,以表征肾活检后6个月以上CKAP4的预测准确性。通过Cox比例风险模型随访CKAP4和DKD进展之间的风险比。将单变量分析中有统计学意义(P<0.05)的临床病理因素作为协变量纳入多变量Cox比例风险模型分析。采用Kaplan⁃Meier分析评价CKAP4高表达组和CKAP4低表达组肾活检后6个月以上生存的差异。结果:与肿瘤患者癌旁正常肾脏组织相比,CKAP4在DKD患者肾脏组织中表达增高,差异有统计学意义(P<0.001)。CKAP4在不同分期的DKD患者肾组织中表达有差异,其中Ⅱ期与Ⅲ期、Ⅱ期与Ⅳ期、Ⅲ期与Ⅳ期之间的差异均有统计学意义(P均<0.05)。DKD患者CKAP4表达与血清肌酐、尿素氮、24 h尿蛋白呈正相关,与肾小球滤过率估计值、血红蛋白呈负相关。DKD患者中位随访期为2.22年,63例(44.06%)患者出现DKD进展。Pearson相关分析显示,CKAP4表达水平随DKD病理分级的升高而升高(r=0.808,P<0.001)。多因素Cox回归分析显示,CKAP4高表达与DKD进展的风险增加相关(HR=4.120,95%CI:1.730~9.811,P=0.001)。此外,在Kaplan⁃Meier生存分析中,CKAP4高表达组患者的肾脏终点事件的发生率显著高于CKAP4低表达组(P<0.001)。绘制了包括CKAP4分类在内的列线图来预测DKD进展风险(C⁃index:0.689)。结论:来源于尿液细胞外囊泡的CKAP4是DKD患者肾活检后6个月以上疾病进展的独立危险因素。
Objective:To investigate the ability of the histological expression of the key protein cytoskeleton-associated protein 4(CKAP4)in urine extracellular vesicles as a biomarker for the progression of diabetic kidney disease(DKD).Methods:A total of 143 type 2 diabetic patients with biopsy-proven DKD and 10 patients with renal malignant tumors were enrolled from the First Affiliated Hospital of Nanjing Medical University.The expression of CKAP4 in renal tissue was detected by immunohistochemistry.Spearman’s correlation analysis was used to analyze the correlation between CKAP4 expression level and clinical indicators.The area under the receiver operating characteristic(ROC)curve(AUC)was calculated to assess whether CKAP4 expression could effectively distinguish patients with poor renal prognosis.ROC curve analysis was used to determine the best cut-off value of CKAP4 score for renal events(highest Youden’s index).Time-dependent AUC analyses were also performed to characterize the predictive accuracy of CKAP4 beyond 6 months after renal biopsy.In addition,hazard ratios between CKAP4 and DKD progression were calculated using by Cox proportional hazards models.The clinicopathological factors with statistical significance(P<0.05)in univariate analysis were included as covariates in multivariate Cox proportional hazards model analysis.Kaplan-Meier analysis was used to evaluate the difference in survival beyond 6 months after renal biopsy between CKAP4 high-expression and CKAP4 low-expression groups.Results:Compared with the adjacent normal kidney tissues of tumor patients,the expression of CKAP4 in the kidneys of DKD patients was significantly increased(P<0.05).The expression of CKAP4 in renal tissues of DKD patients was different in different stages,and the differences between stageⅡand stageⅢ,stageⅡand stageⅣ,and stageⅢand stageⅣwere statistically significant(P<0.05).The expression level of CKAP4 in DKD patients was positively correlated with serum creatinine,urea nitrogen and 24 h urine protein,while negatively correlated with estimated glomerular filtration rate and hemoglobin.During a median follow-up period of 2.22 years,63 patients(44.06%)had DKD progression.Pearson correlation analysis showed that CKAP4 increased with the increase of pathological grade of DKD(r=0.808,P<0.001).Of note,multivariate Cox regression analysis showed that elevated CKAP4 was associated with an increased risk of DKD progression(HR:4.120,95%CI:1.730-9.811,P=0.001).In addition,in Kaplan-Meier survival analysis,patients with high CKAP4 expression had a significantly higher incidence of renal endpoint events than those with low CKAP4 expression(P<0.001).At the same time,a nomogram was developed including CKAP4 classification to predict the risk of DKD progression(C-index:0.689).Conclusion:Our findings suggest that expression of CKAP4,derived from urine extracellular vesicles,is an independent risk factor for disease progression over 6 months after renal biopsy in DKD patients.
作者
刘畅
钱力
陆放
王子韬
李青
段俗言
吴琳
张波
毛慧娟
梁宏伟
袁杨刚
邢昌赢
LIU Chang;QIAN Li;LU Fang;WANG Zitao;LI Qing;DUAN Suyan;WU Lin;ZHANG Bo;MAO Huijuan;LIANG Hongwei;YUAN Yanggang;XING Changying(Department of Nephrology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;School of Life Sciences and Technology,China Pharmaceutical University,Nanjing 210000,China)
出处
《南京医科大学学报(自然科学版)》
北大核心
2025年第8期1063-1072,1100,共11页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省医学创新团队项目(CXTDA2017011)。
