刘营营 聂明豪 袁颖 杨彬 王岩

【摘要】 目的：研究褐藻素对自发性糖尿病相关性糖尿病肾病（DN）的治疗作用。方法：Goto-Kakizaki（GK）大鼠自由食用含或不含褐藻素的标准饲料13周，Wistar大鼠作为对照。褐藻素对分离的大鼠肾小球系膜细胞（GMC）无细胞毒性。用血糖仪测定空腹血糖水平，用全自动生化分析仪测定尿素氮（BUN）和血肌酐（Cr）水平，用ELISA试剂盒测定尿蛋白水平。用ELISA试剂盒检测肾皮质胶原Ⅳ水平，Western blot法检测肾皮质和肾小球系膜细胞转化生长因子-β1（TGF-β1）、纤维连接蛋白（FN）和肾小球系膜细胞核因子-κB（NF-κB）的表达水平。结果：糖尿病组空腹血糖、尿素氮、血肌酐、尿蛋白水平均较对照组、糖尿病+褐藻素组显著升高（P<0.01）;糖尿病组肾皮质胶原Ⅳ、TGF-β1和FN的表达及肾小球系膜细胞TGF-β1和FN均较对照组、糖尿病+褐藻素组显著升高（P<0.01）;糖尿病组NF-κB p65表达明显高于对照组与糖尿病+褐藻素组（P<0.01）。结论：褐藻素通过减弱NF-κB信号通路的激活，降低高血糖，预防或阻止与自发性糖尿病相关性DN的发展。

【关键词】 褐藻素 NF-κB 自发性糖尿病相关性糖尿病肾病

[Abstract] Objective： To study the therapeutic effect of Fucoidan on spontaneous diabetes-associated diabetic nephropathy （DN）. Method： Goto-Kakizaki （GK） rats were allowed free access to standard rat food with or without fucoidan for 13 weeks， and Wistar rats were selected as controls. Fucoidan did not show any cytotoxicity on glomerular mesangial cells （GMCs） which separated from rat kidneys. Fasting blood glucose levels were measured by blood glucose meter， blood urea nitrogen （BUN） and serum creatinine （Cr） levels were measured by automatic biochemistry analyzer and urine protein levels were measured by ELISA kit. Collagen Ⅳ levels in the renal cortex were measured by ELISA kit， and the expression levels of transforming growth factor-β1 （TGF-β1） and fibronectin （FN） in the renal cortex and GMCs， and nuclear factor-κB （NF-κB） in GMCs were determined by Western blot. Result： The levels of fasting blood glucose， BUN， serum Cr and urine protein in diabetic group were significantly higher than those in the control group and diabetic + Fucoidin group （P<0.01）. The expression of collagen Ⅳ， TGF-β1 and FN in renal cortex and the expression of TGF-β1 and FN in glomerular mesangial cells in diabetic group were significantly higher than those in the control group and diabetic + Fucoidin group （P<0.01）. The expression of NF-κB p65 in diabetic group was significantly higher than those in the control group and diabetic + Fucoidin group （P<0.01）. Conclusion： Fucoidan reduces hyperglycemia and prevents or impedes the development of spontaneous diabetes-related DN by attenuating the activation of NF-κB signaling pathway.

[Key words] Fucoidin NF-κB Spontaneous diabetes-related diabetic nephropathy

First-authors address： Heilongjiang Provincial Hospital， Harbin 150000， China

doi：10.3969/j.issn.1674-4985.2021.03.006

糖尿病腎病（DN）是糖尿病最严重的微血管并发症之一，也是终末期肾功能衰竭的主要原因[1]。TGF-β1的表达水平升高已被认为是DN的标志。核因子-κB（NF-κB）是一种转录因子，通常情况下，通过与抑制蛋白IκB的相互作用，以非活性形式被隔离在细胞质中。在糖尿病条件下，激活的NF-κB移位到细胞核，并触发其目标基因的表达，包括TGF-β1和FN，进一步导致细胞外基质的积累[2]。褐藻素（Fucoidan）是一种海藻提取物，作为一种抗氧化剂、抗癌和抗炎剂已被广泛研究，并已被证明在癌症和炎症中发挥重要作用[3-4]。因此，笔者预期通过本研究，确定褐藻素是否能抑制自发性糖尿病相关性DN的发展。现报道如下。

1 材料与方法

1.1 动物 实验在哈尔滨医科大学动物实验中心进行，并经哈尔滨医科大学伦理中心批准。……