水飞蓟素通过共同抑制TLR4/NF-κB和TNF-α/ROS/P38MAPK通路减轻糖尿病肾病大鼠肾脏损伤的研究

Silymarin jointly inhibits TLR4/NF-κB and TNF-α/ROS/P38mapk pathway reduces renal injury in diabetes nephropathy rats

目的:探讨水飞蓟素通过共同抑制Toll样受体4/核因子-κB(TLR4/NF-κB)和肿瘤坏死因子α/活性氧簇/P38丝裂原活化蛋白激酶(TNF-α/ROS/P38MAPK)通路减轻糖尿病肾病(DN)大鼠肾脏损伤的机制。方法:选取健康SD大鼠50只,按随机数字表法分为对照组、模型组、低剂量水飞蓟素组、中剂量水飞蓟素组、高剂量水飞蓟素组,每组10只。建模成功并治疗8周后,生化分析仪测定各组大鼠空腹血糖(FBG)、血肌酐(Scr)、尿素氮(BUN)、胱抑素C(Cys-C)、β_(2)微球蛋白(β_(2)-MG)、24 h尿蛋白(UTP)水平;计算肾脏指数;试剂盒检测各组大鼠肾组织丙二醛(MDA)、总抗氧化能力(T-AOC)水平;RT-PCR检测各组大鼠TLR4,NF-κB,TNF-α和P38MAPK等mRNA的表达水平。结果:与对照组相比,模型组体质量、T-AOC均降低(P<0.05),肾脏指数和FBG,Scr,BUN,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均升高(P<0.05)。低、中、高剂量水飞蓟素组体质量、T-AOC水平均低于对照组,且均高于模型组(P<0.05);低、中、高剂量水飞蓟素组肾脏指数和FBG,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均高于对照组,且均低于模型组(P<0.05);低、中剂量水飞蓟素组Scr,BUN水平均高于对照组,且低于模型组(P<0.05);高剂量水飞蓟素组Scr,BUN水平均低于模型组(P<0.05),高于对照组,但差异无统计学意义(P>0.05)。结论:水飞蓟素可调控TLR4/NF-κB和TNF-α/ROS/P38MAPK通路,可通过抑制其激活减轻氧化应激、减轻DN大鼠的肾脏损伤。

Objective:To explore the effect of silymarin on Toll like receptor 4(TLR4)/nuclear factor-κB(NF-κB)and tumor necrosis factor(TNF-α)/mechanism of reactive oxygen species(ROS)/P38 mitogen activated protein kinase(P38MAPK)pathway in alleviating renal injury in diabetes nephropathy(DN)rats.Methods:Fifty healthy SD rats were randomly divided into control group,model group,low-dose silymarin group,medium dose silymarin group and high-dose silymarin group,with 10 rats in each group.After 8 weeks of successful modeling and treatment,the fasting blood glucose(FBG),serum creatinine(SCR),urea nitrogen(BUN),Cystatin C(Cys-C),β_(2)microglobulin(β_(2)-MG),and 24 h urine protein(UTP)level.Renal index was calculated.The levels of malondialdehyde(MDA)and total antioxidative capacity(T-AOC)were detected by kit,and the mRNA expression levels of TLR4,NF-κB,TNF-αand P38MAPK were detected by RT-PCR.Results:Compared with the control group,body mass and T-AOC were decreased in the model group(P<0.05),and renal index,FBG,Scr,BUN,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,and P38MAPK mRNA were increased(P<0.05).Body mass and T-AOC levels were lower in the low,medium and high dose silymarin groups than in the control group,and all were higher than in the model group(P<0.05).Renal index,FBG,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA in the low,medium and high dose silymarin groups were higher than those in the control group,and all were lower than those in the model group(P<0.05).Scr and BUN levels in the low and medium dose silymarin groups were higher than those in the control group,and lower than those in the model group(P<0.05).Scr and BUN levels in the high dose silymarin group were lower than those in the model group(P<0.05)and higher than those in the control group,but the differences were not statistically significant(P>0.05).Conclusion:Silymarin can regulate TLR4/NF-κB and TNF-α/ROS/P38mapk pathway can reduce oxidative stress and renal injury in DN rats by inhibiting its activation.