Department of Pediatrics, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Infectious Department, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Apoptosis. 2023 Apr;28(3-4):539-548. doi: 10.1007/s10495-023-01815-7. Epub 2023 Jan 18.
Recently, there has been an increasing clinical incidence of acute kidney injury (AKI), which rapidly declines renal function and leads to massive tubular cell necrosis. Pyroptosis is an inflammatory process of cell death that is more rapid than apoptosis, which is accompanied by a massive release of inflammasome activation. In the study, we aim to explore whether Bufalin regulates the AKI through the pyroptosis pathway.
We have established gentamicin (GM)-induced AKI animal and cell models to simulate the pathological conditions of kidney injury. The expression of renal injury and pyroptosis-related indicators were detected by western blot. PAS staining and IHC staining were used to analyze renal function. CCK-8 assay was performed to detect cell viability after AKI with different treatments. TUNEL staining, flow cytometry and immunofluorescence assays were performed to measure pyroptosis.
After intraperitoneal injection of GM in rats, renal function was significantly decreased, along with a significant increase of damaged and necrotic cells as suggested by renal tubular epithelial tissue sections. In addition, there was an increase in the pyroptosis-related markers expression and pyroptosis-induced cell death. Consistently, studies in vitro found that GM significantly induced pyroptosis and its associated protein expression in NRK52e cells. Whereas, the administration of Bufalin reversed these effects of GM in vivo and in vitro. Further, we found that Nigericin (NLRP3 agonist) could reversed the effects of bufalin on GM-induced pyroptosis.
Bufalin attenuates pyroptosis generated AKI by inhibiting NLRP3 inflammasome.
最近，急性肾损伤（AKI）的临床发病率不断增加，其迅速降低肾功能并导致大量肾小管细胞坏死。细胞焦亡是一种比细胞凋亡更快的细胞死亡炎症过程，伴随着大量炎症小体激活的释放。在研究中，我们旨在探讨蟾毒灵是否通过细胞焦亡途径调节 AKI。
我们建立了庆大霉素（GM）诱导的 AKI 动物和细胞模型，以模拟肾脏损伤的病理条件。通过 Western blot 检测肾损伤和细胞焦亡相关指标的表达。PAS 染色和 IHC 染色分析肾功能。CCK-8 测定不同处理后 AKI 细胞的活力。TUNEL 染色、流式细胞术和免疫荧光法测定细胞焦亡。
GM 腹腔注射大鼠后，肾功能明显下降，肾小管上皮组织切片显示损伤和坏死细胞明显增加。此外，还观察到细胞焦亡相关标志物表达和细胞焦亡诱导的细胞死亡增加。体外研究发现 GM 显著诱导 NRK52e 细胞发生细胞焦亡及其相关蛋白表达。而蟾毒灵在体内和体外均逆转了 GM 的这些作用。进一步研究发现， Nigericin（NLRP3 激动剂）可逆转蟾毒灵对 GM 诱导的细胞焦亡的作用。
蟾毒灵通过抑制 NLRP3 炎症小体减轻 GM 诱导的细胞焦亡引起的 AKI。
