摘要
目的:研究活血地龟汤(HXDGT)对高糖诱导的足细胞铁死亡的调控作用,明确其治疗糖尿病肾脏疾病(DKD)的作用机制。方法:体外常规培养MPC-5足细胞,分为正常组、DKD组、HXDGT组、核转录因子红系2相关因子2(Nrf2)激动剂莱菔硫烷(Sulforaphane)组、Nrf2抑制剂ML385组,免疫荧光染色检测足突蛋白(Podocin)、肾病蛋白(Nephrin)、谷胱甘肽过氧化物酶4(GPX4)、酰基辅酶A合成酶长链家族成员4(ACSL4)、Nrf2、铁转运蛋白1(FPN1)的蛋白表达和定位;Western Blot法检测GPX4、ACSL4、Nrf2、FPN1蛋白的表达;普鲁士蓝染色观察足细胞内铁离子积累情况。结果:与正常组比较,DKD组Podocin、Nephrin、GPX4、Nrf2、FPN1荧光强度减弱,ACSL4荧光强度增强,GPX4、Nrf2、FPN1蛋白表达下调(P<0.01),ACSL4蛋白表达上调(P<0.01),普鲁士蓝染色加深;与DKD组比较,HXDGT组和Sulforaphane组Podocin、Nephrin、GPX4、Nrf2、FPN1荧光强度增强,ACSL4荧光强度减弱,GPX4蛋白表达上调(P<0.01)、Nrf2蛋白表达上调(P<0.05),FPN1蛋白表达上调(P<0.01,P<0.05),普鲁士蓝染色减轻。结论:HXDGT可能通过上调Nrf2蛋白表达,促进FPN1蛋白表达,减轻DKD足细胞铁沉积以减轻铁死亡,从而发挥保护足细胞、治疗DKD的作用。
Objective:To investigate the regulatory effects of Huoxue Digui Decoction(HXDGT) on high glucose-induced ferroptosis in podocytes and to elucidate its therapeutic mechanism in diabetic kidney disease(DKD).Methods:MPC-5 podocytes were cultured in vitro and divided into normal,DKD,HXDGT,Nrf2 activator(Sulforaphane),and nuclear factor erythroid 2-related factor 2(Nrf2) inhibitor(ML385) groups.Immunofluorescence staining was used to detect the expression and localization of podocin,nephrin,glutathione peroxidase 4(GPX4),acyl-CoA synthetase long-chain family member 4(ACSL4),Nrf2,and ferroportin 1(FPN1).Western blot was employed to assess the protein expression of GPX4,ACSL4,Nrf2,and FPN1.Prussian blue staining was performed to evaluate intracellular iron accumulation in podocytes.Results:Compared with the normal group,the DKD group exhibited reduced fluorescence intensities of podocin,nephrin,GPX4,Nrf2,and FPN1,and increased fluorescence intensity of ACSL4.Protein expression levels of GPX4,Nrf2,and FPN1 were significantly downregulated(P<0.01),while ACSL4 was significantly upregulated(P<0.01),accompanied by enhanced Prussian blue staining.Compared with the DKD group,both the HXDGT and Sulforaphane groups showed enhanced fluorescence intensities of podocin,nephrin,GPX4,Nrf2,and FPN1,and reduced ACSL4 fluorescence.Additionally,GPX4 expression was significantly upregulated(P<0.01),Nrf2 expression was upregulated(P<0.05),and FPN1 expression was elevated(P<0.01,P<0.05),with reduced Prussian blue staining.Conclusion:HXDGT may exert its protective effects on podocytes and therapeutic action against DKD by upregulating Nrf2 protein expression,promoting FPN1 protein expression,alleviating iron accumulation,and thereby inhibiting ferroptosis.
作者
李冰
王学艺
韩佳瑞
王慧丽
王晨辉
LI Bing;WANG Xueyi;HAN Jiarui;WANG Huili;WANG Chenhui(Second Clinical College of Henan University of Chinese Medicine,Zhengzhou 450002,China;Department of Nephrology,Henan Province Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)
出处
《世界中医药》
北大核心
2025年第5期738-745,共8页
World Chinese Medicine
基金
国家自然科学基金面上项目(82205082)
河南省中医药科学研究专项重点课题(20-21ZY1003)
河南省重点研发与推广专项(科技攻关)项目(202102310171,202102310505)
河南省中医药拔尖人才培养项目(2019ZYBJ17)
全国中医药创新骨干人才培训项目。
