Trauma Center, Zhuzhou Hospital Affiliated to Xiangya Medical College, Central South University, Zhuzhou, Hunan, China.
Department of Emergency, Zhuzhou Hospital Affiliated to Xiangya Medical College, Central South University, Zhuzhou, Hunan, China.
Kaohsiung J Med Sci. 2024 Sep;40(9):789-800. doi: 10.1002/kjm2.12883.
We investigated the potential correlation between miR-223 and NAcHT, LRR, and PYd domain-containing protein 3 (NLRP3) in the context of renal ischemia-reperfusion injury (RIRI), which is a leading cause of acute renal failure with significant mortality rates. Additionally, miR-223 has been implicated in renal inflammation, further highlighting its relevance to this study. C57BL/6 male mice were used as RIRI models. After successful modeling, pathological examinations and serum creatinine and miR-223 levels were tested. Pro-inflammatory cytokine (IL-1β, IL-6, IL-8, NLPR3, TLR4) expression was detected in mice by western blot (kidney tissue) and enzyme-linked immunosorbent assay (serum). HK-2 cells were used for in vitro experiments. A hypoxia/reoxygenation (H/R) model was used, and miR-223 and pro-inflammatory cytokine levels were detected using PCR and western blot assays, respectively. A dual-luciferase reporter assay was conducted to confirm the binding of miR-223 to NLPR3. Next, NLRP3 was knocked down to determine whether the anti-inflammatory function of miR-223 is dependent on NLRP3. MiR-223 expression was lower in RIRI mice than in the sham operation group. The level of miR-223 negatively correlated with serum creatinine levels and the severity of tubule injury. Increased proinflammatory cytokine levels in RIRI mice were observed. In vitro, miR-223 alleviated the inflammatory response in H/R treated cells by inhibiting proinflammatory cytokines. Dual-luciferase reporter and western blot assays confirmed the binding of miR-223 to NLRP3. NLRP3 knockdown reversed the anti-inflammatory effects of miR-223 in HK-2 cells. MiR-223 plays an anti-inflammatory role in RIRI by targeting NLRP3 to repress pro-inflammatory factors.
我们研究了 miR-223 与 NAcHT、LRR 和富含 PYd 结构域的蛋白 3（NLRP3）之间的潜在相关性，这在肾缺血再灌注损伤（RIRI）中是一个主要原因，它是具有高死亡率的急性肾衰竭的主要原因。此外，miR-223 与肾炎症有关，这进一步突出了它与本研究的相关性。C57BL/6 雄性小鼠被用作 RIRI 模型。成功建模后，进行病理检查和血清肌酐及 miR-223 水平检测。通过 Western blot（肾组织）和酶联免疫吸附试验（血清）检测小鼠促炎细胞因子（IL-1β、IL-6、IL-8、NLRP3、TLR4）的表达。使用体外实验的 HK-2 细胞。使用缺氧/复氧（H/R）模型，分别通过 PCR 和 Western blot 检测 miR-223 和促炎细胞因子水平。进行双荧光素酶报告基因实验以确认 miR-223 与 NLRP3 的结合。接下来，敲低 NLRP3 以确定 miR-223 的抗炎功能是否依赖于 NLRP3。与假手术组相比，RIRI 小鼠的 miR-223 表达降低。miR-223 的表达水平与血清肌酐水平和肾小管损伤的严重程度呈负相关。在 RIRI 小鼠中观察到促炎细胞因子水平升高。在体外，miR-223 通过抑制促炎细胞因子减轻 H/R 处理细胞的炎症反应。双荧光素酶报告基因和 Western blot 实验证实了 miR-223 与 NLRP3 的结合。NLRP3 敲低逆转了 miR-223 在 HK-2 细胞中的抗炎作用。miR-223 通过靶向 NLRP3 抑制促炎因子在 RIRI 中发挥抗炎作用。
