Integrative analysis of transcriptome and metabolome provide new insights into mechanisms of Capilliposide A against cisplatin-induced nephrotoxicity

Objective: Cisplatin (CDDP) has been widely used for chemotherapy against tumours. However,the nephrotoxicity has limited its clinical use. Here, we reported a novel compound, Capilliposide A (CPS-A), to exhibit therapeutic effects on CDDP-induced acute kidney injury (AKI)and explored its potential mechanisms via transcriptome and metabolome. Materials and methods: HK-2 cells were treated with CPS-A, after which cell viability, apoptosis and inflammation were investigated. A mouse model of AKI was constructed by single injection of CDDP in vivo. The renal function and morphology and mitochondrial function were assessed by pathological section and transmission electron microscope (TEM). Transcriptomics and metabolomics are used to explore possible mechanisms which was later verified in vitro. Results: CPS-A administration improved the survival rates of HK-2 cells with a significant decrease in the expression of KIM-1, NGAL, IL-6, IL-8 and IL-1ß. In vivo results also suggested that CPS-A attenuates CDDP-induced kidney injury by reducing serum creatinine (Cr) and blood urea nitrogen (BUN) levels. Furthermore, TEM also showed the improvement of mitochondrial ultrastructure both in vivo and vitro. Transcriptomics analysis of the mice's renal cortex indicated the expression of ATF4 and CHOP were upregulated, which was further validated by qPCR and Western blotting in vitro. Integrative analysis of transcriptome and metabolome indicated that L-Leucine enriched in Valine, leucine and isoleucine degradation might be potential targets. Conclusions: CPS-A can effectively regulate endogenous metabolites associated with amino acid metabolism and ameliorate apoptosis and oxidative stress in CDDP-induced AKI by reducing endoplasmic reticulum stress. Overall design: To explore possible mechanism of CPS-A against cisplatin-induced acute kidney injury,Transcriptomics of mouse's kidney is conducted.

研究目的：顺铂（Cisplatin, CDDP）是临床广泛应用的肿瘤化疗药物，但其引发的肾毒性极大限制了其临床应用。本研究报道了一种新型化合物——海胆苷A（Capilliposide A, CPS-A），其对顺铂诱导的急性肾损伤（Acute Kidney Injury, AKI）具有明确治疗作用，并通过转录组学与代谢组学技术探究其潜在作用机制。
材料与方法：采用CPS-A处理HK-2细胞，随后检测细胞活力、细胞凋亡及炎症因子表达水平。体内实验通过单次注射顺铂构建AKI小鼠模型，利用病理切片与透射电子显微镜（Transmission Electron Microscope, TEM）评估肾功能、肾脏形态及线粒体功能。采用转录组学与代谢组学分析潜在作用机制，并在体外实验中进行验证。
结果：CPS-A处理可显著提升HK-2细胞存活率，并下调KIM-1、NGAL、IL-6、IL-8及IL-1β的表达水平。体内实验结果显示，CPS-A可通过降低血清肌酐（Creatinine, Cr）与血尿素氮（Blood Urea Nitrogen, BUN）水平，减轻顺铂诱导的肾损伤。透射电子显微镜观察证实，CPS-A可改善体内外模型中的线粒体超微结构损伤。小鼠肾皮质转录组学分析显示，ATF4与CHOP的表达水平显著上调，该结果通过体外qPCR与Western blotting实验得到验证。转录组与代谢组联合分析表明，富集于缬氨酸、亮氨酸与异亮氨酸降解通路的L-亮氨酸可能是潜在的作用靶点。
结论：CPS-A可有效调控与氨基酸代谢相关的内源性代谢物，通过减轻内质网应激，改善顺铂诱导的急性肾损伤中的细胞凋亡与氧化应激反应。
整体实验设计：为探究CPS-A对抗顺铂诱导急性肾损伤的潜在作用机制，本研究开展了小鼠肾脏组织的转录组学测序分析。