Department of nephrology, West China Hospital, School of Clinic Medicine, Sichuan University, 610041, Chengdu, Sichuan, China.
Department of Cardiovascular diseases, West China Hospital, School of Clinic Medicine, Sichuan University, 610041, Chengdu, Sichuan, China.
BMC Nephrol. 2023 Feb 2;24(1):25. doi: 10.1186/s12882-023-03066-9.
Contrast-induced acute kidney injury (CI-AKI) is the third most common cause of hospital-acquired renal failure. However, there is no effective treatment of CI-AKI, and its mechanism is unknown. Interestingly, atorvastatin has been reported to be effective in renal injury. Therefore, the aim of this study was to explore the effect and possible molecular mechanism of atorvastatin in CI-AKI.
On the CI-AKI in vitro model, rat tubular epithelial cells (NRK-52E) were treated with 18 mg I/ml meglumine diatrizoate (MEG) and then pretreated with atorvastatin. pcDNA3.1-TLR4 treatment was performed to overexpress toll-like receptor 4 (TLR4) in NRK-52E cells. Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) kits were used to detect NRK-52E cell viability as well as LDH release in each group, respectively; qRT-PCR to determine mRNA expression of TLR4 in cells; western blot to detect protein expression levels of pyroptosis-related proteins (NLRP3, caspase-1, ASC, and GSDMD) and TLR4/MyD88/NF-κB signaling pathway-related proteins (TLR4, MyD88, NF-κBp65, and p-NF-κB p65) in cells.
MEG treatment significantly inhibited the viability of NRK-52E cells, increased pro-inflammatory factor levels and promoted pyroptosis, representing successful establishment of a rat tubular epithelial cell (NRK-52E) CI-AKI in vitro model. Notably, atorvastatin increased the activity of MEG-treated NRK-52E cells and alleviated cell injury in a concentration-dependent manner. In addition, atorvastatin significantly down-regulated the expression of TLR4 in MEG-treated NRK-52E cells. However, overexpression of TLR4 inhibited the effects of atorvastatin on increasing cell viability, alleviating cell injury, reducing pro-inflammatory factors (IL-1β, IL-6, and TNF-α) levels, and inhibiting apoptosis (by down-regulating the expression of NLRP3, caspase-1, ASC, and GSDMD). Furthermore, atorvastatin also inhibited the expression of TLR4/MyD88/NF-κB pathway-related proteins (TLR4, MyD88, and p-NF-κB p65).
Atorvastatin can attenuate CI-AKI through increasing the activity of MEG-treated renal tubular epithelial cells, relieving cell injury, as well as inhibiting pyroptosis and inflammation. More importantly, the mechanism was achieved by inhibiting the TLR4//MyD88/NF-κB signaling pathway.
对比剂诱导的急性肾损伤（CI-AKI）是医院获得性肾衰竭的第三大常见原因。然而，目前还没有有效的治疗方法，其机制尚不清楚。有趣的是，阿托伐他汀已被报道对肾损伤有效。因此，本研究旨在探讨阿托伐他汀对 CI-AKI 的作用及其可能的分子机制。
在体外 CI-AKI 模型中，用 18mg/ml 葡甲胺二碘酞酸盐（MEG）处理大鼠肾小管上皮细胞（NRK-52E），然后用阿托伐他汀预处理。用 pcDNA3.1-TLR4 处理过表达 toll 样受体 4（TLR4）的 NRK-52E 细胞。细胞计数试剂盒-8（CCK-8）和乳酸脱氢酶（LDH）试剂盒分别用于检测各组 NRK-52E 细胞活力和 LDH 释放；qRT-PCR 检测细胞中 TLR4 的 mRNA 表达；western blot 检测细胞中焦亡相关蛋白（NLRP3、caspase-1、ASC 和 GSDMD）和 TLR4/MyD88/NF-κB 信号通路相关蛋白（TLR4、MyD88、NF-κBp65 和 p-NF-κB p65）的蛋白表达水平。
MEG 处理显著抑制 NRK-52E 细胞活力，增加促炎因子水平并促进焦亡，成功建立了大鼠肾小管上皮细胞（NRK-52E）体外 CI-AKI 模型。值得注意的是，阿托伐他汀以浓度依赖性方式增加 MEG 处理的 NRK-52E 细胞的活性并减轻细胞损伤。此外，阿托伐他汀显著下调 MEG 处理的 NRK-52E 细胞中 TLR4 的表达。然而，TLR4 的过表达抑制了阿托伐他汀增加细胞活力、减轻细胞损伤、降低促炎因子（IL-1β、IL-6 和 TNF-α）水平和抑制凋亡（通过下调 NLRP3、caspase-1、ASC 和 GSDMD 的表达）的作用。此外，阿托伐他汀还抑制 TLR4/MyD88/NF-κB 通路相关蛋白（TLR4、MyD88 和 p-NF-κB p65）的表达。
阿托伐他汀通过增加 MEG 处理的肾小管上皮细胞的活性、减轻细胞损伤以及抑制焦亡和炎症来减轻 CI-AKI。更重要的是，该机制是通过抑制 TLR4//MyD88/NF-κB 信号通路来实现的。
