针对小鼠肾缺血/再灌注损伤的促红细胞生成素衍生肽环 B 表面肽与 Caspase-3 siRNA 的有效治疗及转录谱分析。
Potent Therapy and Transcriptional Profile of Combined Erythropoietin-Derived Peptide Cyclic Helix B Surface Peptide and Caspase-3 siRNA against Kidney Ischemia/Reperfusion Injury in Mice.
机构信息
Renal Group, Basic Medical Research Centre, Nantong University, Nantong, China (Y.W., Y.Z., A.L.); Leicester-Nantong Joint Institute of Kidney Science, Department of Nephrology, Affiliated Hospital of Nantong University, Nantong, China (W.C., H.W., Y.F., B.Y.); Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China (C.Y., T.Z.); Shanghai Key Laboratory of Organ Transplantation, Shanghai, China (C.Y., T.Z.); and Renal Group, Department of Cardiovascular Sciences, University of Leicester, University Hospitals of Leicester, Leicester, United Kingdom (Y.W., B.Y.).
Renal Group, Basic Medical Research Centre, Nantong University, Nantong, China (Y.W., Y.Z., A.L.); Leicester-Nantong Joint Institute of Kidney Science, Department of Nephrology, Affiliated Hospital of Nantong University, Nantong, China (W.C., H.W., Y.F., B.Y.); Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China (C.Y., T.Z.); Shanghai Key Laboratory of Organ Transplantation, Shanghai, China (C.Y., T.Z.); and Renal Group, Department of Cardiovascular Sciences, University of Leicester, University Hospitals of Leicester, Leicester, United Kingdom (Y.W., B.Y.)
出版信息
J Pharmacol Exp Ther. 2020 Oct;375(1):92-103. doi: 10.1124/jpet.120.000092. Epub 2020 Aug 5.
Cause-specific treatment and timely diagnosis are still not available for acute kidney injury (AKI) apart from supportive therapy and serum creatinine measurement. A novel erythropoietin-derived cyclic helix B surface peptide (CHBP) protects kidneys against AKI with different causes, but the underlying mechanism is not fully defined. Herein, we investigated the transcriptional profile of renoprotection induced by CHBP and its potential synergistic effects with siRNA targeting caspase-3, an executing enzyme of apoptosis and inflammation (CASP3siRNA), on ischemia/reperfusion (IR)-induced AKI. Utilizing a mouse model with 30-minute renal bilateral ischemia and 48-hour reperfusion, the renoprotection of CHBP or CASP3siRNA was demonstrated in renal function and structure, active caspase-3 and HMGB1 expression. Combined treatment of CHBP and CASP3siRNA further preserved kidney structure and reduced active caspase-3 and HMGB1. Furthermore, differentially expressed genes (DEGs) were identified with fold change >1.414 and < 0.05. In IR kidneys, 281 DEGs induced by CHBP were mainly involved in promoting cell division and improving cellular function and metabolism (upregulated signal transducer and activator of transcription 5B and solute carrier family 22 member 7). The additional administration of CASP3siRNA caused 504 and 418 DEGs in IR + CHBP kidneys with or without negative control small-interfering RNA, with 37 genes in common. These DEGs were associated with modulated apoptosis and inflammation (upregulated BCL6, SLPI, and SERPINA3M) as well as immunity, injury, and microvascular homeostasis (upregulated complement factor H and GREM1 and downregulated ANGPTL2). This proof-of-effect study indicated the potent renoprotection of CASP3siRNA upon CHBP at the early stage of IR-induced AKI. Underlying genes, BCL6, SLPI, SERPINA3M, GREM1, and ANGPTL2, might be potential new biomarkers for clinical applications. SIGNIFICANCE STATEMENT: It is imperative to explore new strategies of cause-specific treatment and timely diagnosis for acute kidney injury (AKI). CHBP and CASP3siRNA synergistically protected kidney structure after 48-hour ischemia/reperfusion-induced AKI with reduced injury mediators CASP3 and high mobility group box 1. CHBP upregulated cell division-, function-, and metabolism-related genes, whereas CASP3siRNA further regulated immune response- and tissue homeostasis-associated genes. Combined CHBP and CASP3siRNA might be a potent and specific treatment for AKI, and certain dysregulated genes secretory leukocyte peptidase inhibitor and SERPINA3M could facilitate timely diagnosis.
除了支持性治疗和血清肌酐测量外，急性肾损伤（AKI）仍然缺乏特异性治疗和及时诊断。一种新型的促红细胞生成素衍生的环螺旋 B 表面肽（CHBP）可预防不同原因引起的 AKI，但其潜在的机制尚未完全确定。在此，我们研究了 CHBP 诱导的肾保护的转录谱及其与靶向半胱天冬酶-3（凋亡和炎症的执行酶）的 siRNA（CASP3siRNA）的协同作用对缺血/再灌注（IR）诱导的 AKI 的潜在协同作用。利用 30 分钟双侧肾缺血和 48 小时再灌注的小鼠模型，证明了 CHBP 或 CASP3siRNA 在肾功能和结构、活性半胱天冬酶-3 和高迁移率族蛋白 1（HMGB1）表达方面的肾保护作用。联合应用 CHBP 和 CASP3siRNA 进一步保护了肾脏结构，减少了活性半胱天冬酶-3 和 HMGB1。此外，还鉴定了差异表达基因（DEGs），其倍数变化 >1.414 和 <0.05。在 IR 肾脏中，CHBP 诱导的 281 个 DEGs 主要参与促进细胞分裂和改善细胞功能和代谢（上调信号转导子和转录激活子 5B 和溶质载体家族 22 成员 7）。额外给予 CASP3siRNA 导致 IR+CHBP 肾脏中有 504 个和 418 个 DEGs，其中有 37 个基因是共同的。这些 DEGs 与凋亡和炎症的调节（上调 BCL6、SLPI 和 SERPINA3M）以及免疫、损伤和微血管内稳态（上调补体因子 H 和 GREM1 并下调 ANGPTL2）有关。这项效应证明研究表明，在 IR 诱导的 AKI 的早期阶段，CASP3siRNA 在 CHBP 作用下具有强大的肾保护作用。潜在基因 BCL6、SLPI、SERPINA3M、GREM1 和 ANGPTL2 可能是临床应用的潜在新生物标志物。
意义陈述
探索急性肾损伤（AKI）的特异性治疗和及时诊断的新策略至关重要。CHBP 和 CASP3siRNA 在缺血/再灌注诱导的 AKI 后 48 小时协同保护肾脏结构，减少损伤介质 CASP3 和高迁移率族蛋白 1。CHBP 上调与细胞分裂、功能和代谢相关的基因，而 CASP3siRNA 进一步调节与免疫反应和组织内稳态相关的基因。联合应用 CHBP 和 CASP3siRNA 可能是 AKI 的一种有效且特异的治疗方法，某些失调的基因分泌白细胞肽酶抑制剂和 SERPINA3M 可能有助于及时诊断。
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