降糖三黄片对糖尿病肾病大鼠TLR4/NF-κB通路的影响

Effect of Jiangtang Sanhuang Tablets on TLR4/NF-κB Pathway of Diabetic Nephropathy Rats

目的观察降糖三黄片对糖尿病肾病大鼠肾组织Toll样受体4(TLR4)/核因子-κB(NF-κB)通路及单核细胞趋化蛋白^(-1)(MCP^(-1))表达的影响,探讨降糖三黄片对糖尿病肾病的保护作用及其机制。方法将80只SD大鼠随机分为正常组10只,模型组70只。模型组大鼠予腹腔内注射链脲佐菌素(STZ)复制模型,成模大鼠随机分为模型组,降糖三黄片低、中、高剂量组(675,1350,2700 mg·kg^(-1)·d^(-1)),厄贝沙坦组(13.5 mg·kg^(-1)·d^(-1)),正常组和模型组灌服等剂量蒸馏水。给药8周后腹主动脉采血测空腹血糖,收集24 h尿液测定尿蛋白,免疫组化法检测各组肾组织中MCP^(-1)蛋白的分布,Western blot法检测各组肾组织中TLR4、NF-κB(P-p65)蛋白的表达。结果与正常组比较,模型组的空腹血糖、24 h尿蛋白及肾组织TLR4、NF-κB(P-p65)、MCP^(-1)蛋白表达均显著升高,差异有统计学意义(P<0.01)。与模型组比较,降糖三黄片低、中、高剂量组空腹血糖均显著降低,差异有统计学意义(P<0.01)。与模型组比较,降糖三黄片低、中、高剂量组及厄贝沙坦组肾组织TLR4、NF-κB(P-p65)、MCP^(-1)蛋白表达显著降低、尿蛋白排出显著减少,差异均有统计学意义(P<0.05,P<0.01)。结论降糖三黄片通过抑制TLR4/NF-κB通路,下调MCP^(-1)的表达,起到减少尿蛋白排出,延缓DN进展的作用。

Objective To observe the effect of Jiangtang Sanhuang tablets（JTSH） on Toll-like receptor 4（TLR4）/nuclear factor-κB（NF-κB）pathway and the expression of monocyte chemoattractant protein^-1（MCP^-1）of diabetic nephropathy（DN） rats,and to investigate the protective effect and mechanism of JTSH on DN.Methods Eighty SD rats were randomly divided into normal group（n=10）and model group（n=70）.The DN rat model was induced by intraperitoneal injection of streptozocin（STZ）,and then the 70 model rats were randomly divided into DN group,Irbesartan group（13.5 mg·kg^-1·d^-1）,and low-,middle-and high-dose JTSH groups（675,1350,2700mg·kg^-1·d^-1respectively）.The normal group and model group were administered with distilled water.After medication for 8 weeks,the blood was sampled from abdominal aorta and the 24-hour urine was collected.Then we detected fasting blood glucose and 24-hour urine protein.The expression of MCP^-1 in renal tissues was detected by immunohistochemistry,and the expression of TLR4 and NF-κB（P-p65） protein in renal tissues was detected by Western blotting.Results The fasting blood glucose,24-hour urine protein,and renal expression levels of TLR4,NF-κB（P-p65） and MCP^-1 of DN group was significantly higher than those of normal group（P〈0.01）.Compared with the DN group,the fasting blood glucose of the three JTSH groups was significantly lowered（P〈0.05）,and the expression of renal TLR4,NF-κB（P-p65） and MCP^-1 as well as 24-hour urine protein of the three JTSH groups and Irbesartan group were decreased（P〈0.05）.Conclusion JTSH can inhibit TLR4/NF-κB pathway and reduce the expression of MCP^-1 and the excretion of 24-hour urine protein,thus to delay the progression of DN.