基于RNA-Seq鉴定肾小管上皮细胞损伤时趋化因子的表达及其分子机制  

Identification of Chemokines Expression and Molecular Mechanisms of Re⁃nal Tubular Epithelial Cell Injury Based on RNA-Seq

作　　者：刘玺[1] 刘芳[1] 张春华[1] 李春丽 王海澜[3] LIU Xi;LIU Fang;ZHANG Chunhua;LI Chunli;WANG Hailan(Department of Nephrology,Shenzhen Longgang Central Hospital,Shenzhen 518116,Guangdong,China;Depart-ment of Internal Medicine,Shenzhen Longgang Central Hospital,Shenzhen 518116,Guangdong,China;Department of Endocrinology,Longgang Central Hospital of Shenzhen,Shenzhen 518116,Guangdong,China)

机构地区：[1]深圳市龙岗中心医院肾内科,广东深圳518116 [2]深圳市龙岗中心医院内科,广东深圳518116 [3]深圳市龙岗中心医院内分泌科,广东深圳518116

出　　处：《系统医学》2024年第24期41-45,共5页Systems Medicine

基　　金：深圳市龙岗区经济与科技发展专项资金医疗卫生科技计划项目(LGKCYLWS2021000007)。

摘　　要：目的 探索在糖异常情况下诱导的肾小管损伤中趋化因子的表达及其分子机制。方法 选取2020年1月—2021年6月深圳市龙岗中心医院的慢性肾小球肾炎和糖尿病肾病(diabetic kidney disease, DKD)患者的组织样本,各10例。使用不同浓度葡萄糖诱导人肾小管上皮细胞(human kidney 2, HK-2),建立细胞损伤模型,分别用CCK-8分析和流式细胞术检测不同糖浓度对HK-2细胞活力和细胞凋亡的影响;收集细胞RNA样本进行转录组测序分析,筛选差异表达基因并进行实验验证;定量聚合酶链反应和免疫印迹法检测趋化因子CXCL2、CCL20的表达情况。结果 葡萄糖以浓度依赖性的方式降低肾小管上皮细胞活性和促进凋亡。RNA-Seq测序发现,CXCL2和CCL20是不同糖浓度诱导下肾小管细胞表达最为显著的趋化因子。在临床DKD患者活检穿刺标本中,CXCL2和CCL20的mRNA水平均高于慢性肾小球肾炎患者,差异有统计学意义(P均<0.05),蛋白水平也高于慢性肾小球肾炎患者。结论 不同浓度葡萄糖诱导肾小管上皮细胞损伤导致趋化因子表达升高可能介导DKD进展。Objective To explore the expression and molecular mechanism of chemokines in renal tubule injury in‐duced by glucose abnormality.Methods Tissue samples from patients with chronic glomerulo nephritis and diabetic kidney disease(DKD)in Longgang Central Hospital of Shenzhen from January 2020 to June 2021 were selected,in‐cluding 10 patients each.human kidney 2(HK-2)cells were induced with different concentrations of glucose,and the cell injury model was established.CCK-8 analysis and flow cytometry were used to detect the effects of different con‐centrations of glucose on the viability and apoptosis of HK-2 cells.Cell RNA samples were collected for transcriptome sequencing analysis,and differentially expressed genes were screened for experimental verification.The expressions of CXCL2 and CCL20 were detected by quantitative polymerase chain reaction and Western blot.Results Glucose de‐creased the activity of renal tubular epithelial cells and promoted apoptosis in a concentration-dependent manner.RNA-Seq sequencing showed that CXCL2 and CCL20 were the most significant chemokines in renal tubular cells in‐duced by different glucose concentrations.In the biopsy samples of clinical DKD patients,the mRNA levels of CXCL2 and CCL20 were higher than those of patients with chronic glomerulo nephritis,the differences were statistically sig‐nificant(both P<0.05),and the protein levels were also higher than those of patients with chronic glomerulo nephritis.Conclusion The increased expression of chemokines induced by different concentrations of glucose in renal tubular epithelial cells may mediate the progression of DKD.

关 键 词：糖尿病肾病 趋化因子 肾小管损伤 细胞凋亡 

分 类 号：R692[医药卫生—泌尿科学]