Production Of MCP-1,ICAM-1,NF-κB And FN Stimulated By High Glucose In Renal Tubular Epithelial Cells And Intervention Effect Of Shuxuetong

Objective: To observe production of monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1(ICAM-1), nuclear factor-kappaB ( NF-κB) and fibronectin(FN) stimulated by high glucose in cultured human renal tubular epithelial cells(HK-2) and to discuss the role of inflammation in diabetic nephropathy. To investigate the effect and mechanism of Shuxuetong on inflammation in diabetic nephropathy in vitro.Methods: In vitro, cultured human renal tubular epithelial cells were divided into 5 groups: (1)control group(glucose's concentration: 5.5mmol/l) (2)high glucose group (glucose's concentration: 25mmol/l) (3)high glucose + Shuxuetong group(concentration of Shuxuetong: 1μl/ml) (4)high glucose+Shuxuetong group(concentration of Shuxuetong: 5μl/ml) (5) high glucose+Shuxuetong group (concentration of Shuxuetong: 10μl/ml). At 24h, 48h and 72h, MCP-1 secretion and FN secretion were measured in culture supernatants by enzyme-linked immunosorbent assay(ELISA), ICAM-1 level was examined by immunohistochemistry, and NF-κBmRNA expression was analyzed by reverse transcription-polymerase chain reaction(RT-PCR).Results:1. Compared with those in control group, secretion of MCP-1, FN and expression of ICAM-1, NF-κB significantly increased in high glucose group at 24h, 48h and 72h (P<0.01).2. The correlation analysis showed that there was a significant positive correlation among MCP-1, ICAM-1, NF-κB and FN(P<0.01).3. Compared with those in high glucose group, MCP-1 secretion and FN secretion began to decreased in Shuxuetong(1μl/ml) group at 72h, in Shuxuetong(5μl/ml) group at 48h and in Shuxuetong(10μl/ml) group at 24h. ICAM-1 expression and NF-κB expression began to reduce in low concentration of Shuxuetong group(1μl/ml) at 48h and in middle and high concentration of Shuxuetong groups (5μl/ml and 10μl/ml) at 24h.Conclusion:1. High glucose stimulates production of MCP-1, ICAM-1, NF-κB and FN in HK-2, which may help explain the role of inflammation in diabetic nephropathy. 2. MCP-1, ICAM-1, NF-κB and FN are related with each other, suggesting inflammation contributes to the progression of tubulointerstitial fibrosis and the above factors may take part in the pathogenesis of diabetic nephropathy in cooperation.3. Shuxuetong can inhibit inflammation of renal tubular epithelial cells and slow down the fibrosis progression of diabetic nephropathy via suppressing NF-κB expression, MCP-1 and ICAM-1 production and FN deposition stimulated by high glucose.