基于人肾类器官构建顺铂诱导急性肾损伤模型

OBJECTIVE To establish a cisplatin induced acute kidney injury (AKI) model based on human kidney organoids. METHODS (1) Kidney organoids containing various cell types were designed and constructed based on human induced pluripotent stem cells (hiPSC) induction techniques, and the tissue structure and cell types were identified by HE staining and immunofluorescence. (2) Based on the constructed human kidney organoid model, the broad-spectrum anticancer drug cisplatin was used as the experimental drug, and three concentration gradients of 20, 50 and 75 μmol·L^-1 were applied to the human kidney organoid model. The samples were collected 48 h later, and the cell viability was detected by observing the morphological changes of kidney organoids. The expression levels of renal injury factor-1 (Kim-1) and inflammatory cytokine interleukin-8 {IL-8) were detected with real-time quantitative PCR to determine the degree and location of renal injury. RESULTS (1) Histological and immunofluorescence results showed that mature human renal organoids could be produced after iPSC-induced differentiation. The organoids had primitive tubular structure and included various cell types including the proximal tubular tubular, distal tubular tubular, podocytes, interstitial endothelial cells and tubular epithelial cells. (2) Single administration of three concentrations of cisplatin resulted in the destruction of cell morphology and structure, and a large number of tubular structures disappeared. Live/Dead staining showed that renal organoids exhibited dose-dependent apoptosis to cisplatin. With cisplatin at the concentration of 20 μmol·L^-1, cell viability fell below 50%, and approached 0 at a maximum concentration of 75 μmol·L^-1. (3) Kim-1 and IL-8 were significantly increased at different concentrations of cisplatin induced AKI models. CONCLUSION This study has constructed a human kidney organoid model and verified the feasibility of human kidney organoids being used to simulate AKI induced by chemotherapy drugs in vitro. Kim-1 combined with IL-8 is expected to provide data for clinical prediction of the possibility of AKI induced by such drugs and countermeasures against adverse reactions.