Influence of high glucose and mannose binding lectin on interleukin-18 expression in cultured human renal glomerular endothelial cells

Objective To investigate the effects of high glucose and mannose binding lectin(MBL) on expression of interleukin-18 of cultured human renal glomerular endothelial cells(HRGEC).Methods Normal HRGEC cultured in vitro were divided randomly into three groups according to different intervention methods: normal glucose group(5 mmol/L D-glucose),manicol group(5mmol/L D-glucose+25mmol/L manicol)and high glucose group(30mmol/L D-glucose).Real-time PCR and Euzyme-linked Immunosorbent Assay(ELISA) were used to detect IL-18 mRNA expression of HRGEC and protein concentration in supernatant fluid after 72 hours' culture.HRGEC was then randomly divided into two groups: single high glucose group and high glucose+MBL group.After 72 hours' culture with 30mmol/L D-glucose,equivalent volume of 30% MBL deficiency human serum was added into two groups,10μg/mL MBL was only added into high glucose+MBL group,continue the culture for 4 hours.Immunofluorescence was used to detect Menbrane Attacks Complex(MAC)deposition on cell surface,which is the product of Mannose Binding Lectin Complement Pathway(LCP).Real-time PCR and ELISA were used to detect mRNA and protein expression of IL-18 in each group.Results Compared with other groups,the mRNA and protein expression of IL-18 in high glucose group was apparently increased(P0.05).MAC was confirmed obviously deposition on cell surface in high glucose+MBL group.Both mRNA and protein expression of IL-18 in high glucose + MBL group were significantly higher(P0.05) compared with single high glucose group.Conclusion High glucose can bring inflammatory factors' overexpression of HRGEC;high glucose together with MBL can bring LCP activation and promote inflammatory status of diabetic nephropathy.