The Effect Of Danshen Injection On TLR4/NF-κB Signaling Pathway In Renal Tubular Epithelial Cells Induced By High Glucose

AIM To determine the effect of Danshen injection on TLR4/NF-κB signaling pathway in the renal tubular of the rat induced by the high glucose environment in vivo and in vitro,and identify the protective mechanism of Danshen injection in the diabetic nephropathy.METHODS The cell viability of NRK-52 E induced by high glucose was determined by MTT assay.and the Western blot was used to examine the expression of TLR4 in NRK-52 E induced by high glucose in the first chapter.In the second chapter,the effect of Danshen injection on TLR4/ NF-κB signaling in NRK-52 E induced by high glucose were determined by the expression and distribution of TLR4,My D88,NF-κB p65 and p-p65 with the method of Western blot and immunofluorescence.In the last chapter the SD rats were separated to four groups.The rats in the model group,the metformin group and the Danshen injection group were treated with intraperitoneal injection of streptozotocin and high glucose and fat diets after normal feed for a week,after the diabetic rats were established,then treatment with saline or metformin or Danshen injection separately.the rats in control group feed with normal diet and injected with saline of the same dose.After administration for six weeks,the rats were sacrificed and the section of rats kidneys were examined by H&E staining,immunohistochemical staining and Western blots to investigate the pathological changes as well as the expression and distribution of TLR4,My D88 and p65 in each group.RESULTS The expression of TLR4 was significantly up-regulated with the increase of glucose concentration to 50 mmol/L.Treated with the 80 mmol/L glucose for 24 h,the expression of TLR4 in NRK-52 E showed significant difference compared with the normal group..Danshen injection can alleviate the inhibitory effect of high glucose on the proliferation of NRK-52 E cells,and dose-dependently reduce the abnormal up-regulation of TLR4 and My D88,and inhibit the phosphorylation and nuclear translocation of NF-κB p65 protein.In vivo experiments showed that Danshen injection can reduce renal tubular lesions in diabetic rats.The expression of TLR4,My D88 and NF-κB p-p65 protein in the model group was significantly increased(P<0.01),while the TLR4,My D88 the eand NF-κB p-p65 expression in metformin group and Danshen injection group was significantly reduced compared with that of model group.CONCLUSION Danshen injection can significantly reduce the expression of TLR4 and My D88 protein and inhibit NF-κB p65 phosphorylation and nuclear translocation in renal tubule of diabetic rat in vivo and NRK-52 E cells induced by high glucose in vitro,and reduce the inflammation of kidney tissue in diabetic rats.The results showed that Danshen injection can effectively down-regulate the abnormal activation of TLR4/NF-κB signaling pathway in tubule of diabetic rats,which may be one of the mechanisms of Danshen injection to improve diabetic nephropathy.