STM2457减轻顺铂诱导的高尿酸下肾小管细胞损伤

STM2457 reduces cisplatin-induced renal tubular epithelial cells injury in the high uric acid setting

目的观察小鼠在高尿酸血症基础上使用顺铂诱导急性肾损伤(AKI)对于肾脏病变的影响情况以及探究甲基转移酶-3(METTL3)在其中发挥的作用。方法将C57BL/6小鼠随机分成对照组、高尿酸血症组、AKI组和高尿酸血症+AKI复合模型组。通过灌胃氧嗪酸钾+腺嘌呤21 d建立高尿酸血症模型,腹腔注射顺铂3 d建立AKI模型,检测血肌酐、尿素氮、血尿酸、HE、Masson病理染色,探究不同模型对小鼠肾脏损伤情况。Western blot检测α-平滑肌蛋白(α-SMA)、肾损伤分子-1、METTL3蛋白表达变化;qPCR检测白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)、IL-1β等mRNA变化。m6A检测试剂盒检测小鼠总体甲基化水平。结果与单纯顺铂引起的AKI模型组相比,高尿酸血症和AKI复合模型组的血肌酐、尿素氮、血尿酸均明显升高。病理染色、Western blot和qPCR也显示复合模型组肾脏纤维化加重,METTL3表达水平升高。m6A检测结果说明实验组小鼠的总体甲基化水平升高且复合模型组明显高于单纯AKI组。体外实验中,使用METTL3特异性抑制剂STM2457则能够有效减轻肾小管上皮细胞的损伤以及降低炎症。结论顺铂能够加重高尿酸引起的肾脏炎症和纤维化;STM2457对于高尿酸基础上顺铂诱导的细胞损伤具有治疗作用,METTL3可能是治疗有关肾脏损伤的潜在靶点。

Objective To observe the effect of Methyltransferase like 3(METTL3)in cisplatin-induced acute kidney injury(AKI)in mice with hyperuricemia.Methods C57BL/6 mice were randomly divided into control group,hyperuricemia group,AKI group,and hyperuricemia+AKI group.The hyperuricemia model was established by injecting with gavage potassium oxyzinate and adenine for 21 days,and the AKI model was established by intraperitoneal injection of cisplatin for 3 days.Serum creatinine,urea nitrogen,blood uric acid,the pathological staining of HE and Masson were detected.The expression of alpha-smooth muscle actin(α-SMA),Kim-1 and METTL3 proteins were detected by Western blot.qPCR detected mRNA changes such as interleukin-6(IL-6),tumor necrosis factor(TNF-α),and IL-1β.The m6A assay detected overall methylation levels in mice.Results Compared with the AKI group alone,the serum creatinine,urea nitrogen and blood uric acid in the hyperuricemia+AKI group significantly increased.Pathological staining,Western blot,and qPCR also showed worsening of renal fibrosis and elevated METTL3 expression in the composite model group.The m6A test results showed that the overall methylation level of mice in the experimental group increased,and the composite model group was significantly higher than that in the AKI group.In vitro experiments,the METTL3-specific inhibitor STM2457 was given to effectively reduce the injury of renal tubular epithelial cells and reduce the level of inflammation and fibrosis.Conclusion Cisplatin can exacerbate renal inflammation and fibrosis caused by high uric acid;STM2457 has therapeutic effects in hyperuricemic nephropathy and cisplatin-induced acute kidney injury,and METTL3 may be a potential target for the treatment of related kidney injury.