Wu Fan, Dong Xiu-Juan, Li Yan-Yan, Zhao Yan, Xu Qiu-Lin, Su Lei
Southern Medical UniversityGuangzhou 510515, Guangdong, China; Department of Nephrology, The Third Hospital of Zhengzhou 450000Zhengzhou, Henan, China.
Department of Hematology, The Third Hospital of Zhengzhou 450000 Zhengzhou, Henan, China.
Int J Clin Exp Pathol. 2015 Nov 1;8(11):14409-16. eCollection 2015.
Acute kidney injury (AKI) is a common and increasingly encountered complication in hospitalized patients with critical illness in intensive care units (ICU). According to the etiology, Sepsis-induced AKI (SAKI) is a leading contributor to AKI and significantly has very poor prognosis, which might be related to the late detection when the elevation of BUN and serum creatinine (SCr) is used. Many genes are up-regulated in the damaged kidney with the corresponding protein products appearing in plasma and urine. Some of these are candidate biomarkers for more timely diagnosis of SAKI. Therefore, extensive research efforts over this past decade have been directed at the discovery and validation of novel SAKI biomarkers to detect injury prior to changes in kidney function, a number of serum and urinary proteins, including NGAL, KIM-1, cystatin-C, IL-18, and L-FABP, have been identified for predicting SAKI before a rise in BUN and serum creatinine in several experimental and clinical trainings. Unfortunately, an ideal biomarker of SAKI with highly sensitivity and specificity has not been identified yet. Recent progresses in quantitative proteomics have offered opportunities to discover biomarkers for SAKI. In the present study, kidney tissue samples from SAKI mice were analyzed by two-dimensional differential gel electrophoresis (2D-DIGE), and 4 up-regulated proteins, which were actin (ACTB), myosin regulatory light chain 12B (MYL12B), myosin regulatory light polypeptide 9 (MYL9), and myosin regulatory light chain 12A (MYL12A) were identified by matrix assisted laser desorption ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). Among all the varied proteins, MYL12B was validated by western blot. Interestingly, there was no change between the SAKI and control kidney tissues, however, phosphorylated MYL12B was detected to be consistent with the proteomics data. Furthermore, phosphorylated MYL12B was found similarly to be increased in SAKI plasma, while MYL12B was changeless in plasma of control group. Taking together, phosphorylated MYL12B may be employed as a potential plasma biomarker for the early diagnosis of SAKI.
急性肾损伤(AKI)是重症监护病房(ICU)中危重症住院患者常见且日益增多的并发症。根据病因,脓毒症诱导的急性肾损伤(SAKI)是急性肾损伤的主要原因,其预后极差,这可能与使用血尿素氮(BUN)和血清肌酐(SCr)升高进行晚期检测有关。许多基因在受损肾脏中上调,相应的蛋白质产物出现在血浆和尿液中。其中一些是更及时诊断SAKI的候选生物标志物。因此,在过去十年中,大量研究致力于发现和验证新型SAKI生物标志物,以便在肾功能改变之前检测损伤,在一些实验和临床研究中,已经确定了多种血清和尿液蛋白,包括中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、肾损伤分子-1(KIM-1)、胱抑素-C、白细胞介素-18(IL-18)和肝型脂肪酸结合蛋白(L-FABP),可在BUN和血清肌酐升高之前预测SAKI。不幸的是,尚未确定一种具有高灵敏度和特异性的理想SAKI生物标志物。定量蛋白质组学的最新进展为发现SAKI生物标志物提供了机会。在本研究中,通过二维差异凝胶电泳(2D-DIGE)分析了SAKI小鼠的肾组织样本,并通过基质辅助激光解吸电离飞行时间/飞行时间质谱(MALDI-TOF/TOF MS)鉴定了4种上调蛋白,即肌动蛋白(ACTB)、肌球蛋白调节轻链12B(MYL12B)、肌球蛋白调节轻多肽9(MYL9)和肌球蛋白调节轻链12A(MYL12A)。在所有变化的蛋白质中,通过蛋白质印迹法验证了MYL12B。有趣的是,SAKI肾组织与对照肾组织之间没有变化,然而,检测到磷酸化的MYL12B与蛋白质组学数据一致。此外,发现磷酸化的MYL12B在SAKI血浆中同样增加,而在对照组血浆中MYL12B无变化。综上所述,磷酸化的MYL12B可作为SAKI早期诊断的潜在血浆生物标志物。