摘要
目的 探讨萝卜硫素(sulforaphane,SFN)通过调控AMPK/SIRT1/NF-κB途径保护糖尿病小鼠肾损伤的治疗机制。方法 30只C57BL/6小鼠随机分为正常组10只和实验组20只,后者通过腹腔注射50 mg/kg链脲佐菌素制备糖尿病小鼠模型;造模成功的小鼠分为模型组和萝卜硫素组(每天萝卜硫素0.5 mg/kg)各10只。每周称重并测量空腹血糖,连续给药12周后,收集血清和肾脏组织,检测肾损伤指标、炎症因子及AMPK/SIRT1/NF-κB信号通路的蛋白水平,观察肾脏病理损伤情况。高糖处理HEK-293T细胞复制肾细胞损伤模型,加入AMPK抑制剂验证SFN是否通过激活AMPK/SIRT1途径发挥抗炎作用。结果 给药12周后,SFN组小鼠空腹血糖降至7.5 mmol/L,胰岛素升至0.82 ng/mL,肾脏病理损伤明显改善,血清中肌酐、尿素氮、24 h内尿蛋白含量及促炎因子(TNF-α、IL-1β和IL-6)、p-p65蛋白水平降低,p-AMPK和SIRT1的蛋白表达水平显著增加,与模型组差异均有统计学意义(P<0.05)。体外实验表明,SFN改善高糖诱导的HEK-293T细胞炎症,激活AMPK/SIRT1信号通路并降低p65磷酸化蛋白表达水平;而AMPK抑制剂削弱了SFN的抗炎效果。结论 萝卜硫素通过激活AMPK/SIRT1信号通路并抑制NF-κB信号通路降低肾脏炎症水平,有效缓解糖尿病小鼠肾损伤。
Objective To investigate the therapeutic mechanism of sulforaphane(SFN)to protect against kidney injury in diabetic mice by regulating AMPK/SIRT1/NF-κB pathway.Methods A total of 30 C57BL/6 mice were randomly divided into a normal control group(10 mice)and an experimental group(20 mice).The latter group was administered a diabetic mouse model via intraperitoneal injection of 50 mg/kg streptozotocin(STZ);the successful model mice were then divided into the model group and the sulforaphane group(each with 10 mice),with the latter receiving daily doses of 0.5 mg/kg sulforaphane.Body weight was measured weekly,and fasting blood glucose was assessed.After 12 weeks of continuous dosing,serum and kidney tissues were collected to test for kidney injury markers,inflammatory factors,and protein levels of the AMPK/SIRT1/NF-κB signaling pathway.Renal pathological damage was observed.High-glucose treated HEK-293T cells were used to replicate the renal cell injury model,and an AMPK inhibitor was added to verify whether SFN exerts anti-inflammatory effects through the activation of the AMPK/SIRT1 pathway.Results In the SFN group,fasting blood glucose levels were reduced to 7.5 mmol/L,insulin levels increased to 0.82 ng/mL,and renal pathological damage was significantly improved.Serum creatinine,blood urea nitrogen,24-hour urinary protein excretion,and pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6),as well as the protein levels of p-p65,were decreased.In contrast,the protein expression levels of p-AMPK and SIRT1 were significantly increased,with all differences being statistically significant compared to the model group(P<0.05).In vitro experiments indicated that SFN ameliorated high-glucose-induced inflammation in HEK-293T cells by activating the AMPK/SIRT1 signaling pathway and reducing the expression levels of phosphorylated p65 protein.The anti-inflammatory effects of SFN were attenuated by the AMPK inhibitor.Conclusions SFN reduces kidney inflammation by activating AMPK/SIRT1 signaling pathway and inhibiting NF-κB si
作者
缪蒙爱
郑楠
杨敏
MIAO Meng’ai;ZHENG Nan;YANG Min(Department of Critical Care Medicine,People s Hospital of Pingyang,Wenzhou 325400,China;School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325000,China;Department of Orthopedics,Pingyang Hospital of Traditional Chinese Medicine,Wenzhou 325400,China)
出处
《健康研究》
CAS
2024年第5期553-557,F0003,共6页
Health Research
基金
浙江省基础公益研究计划(LY18C020006)
温州市级基础性科研项目(Y2023373)。