四川大学学报（医学版）2011，Vol.42Issue(1)：90-94,5.
高糖和MBL途径的补体激活对人肾小球内皮细胞表达IL-6、TNF-α的影响
Influenee of High G lucnse and Mannose Binding Lectin Complement Pathwav Activation to IL-6 and TNF-α's Expression by Human Renal Glomerular Endothelial Cells
摘要
Abstract
Objective To investigate the effect of high glucose and mannose binding lectin (MBL) complement pathway activation's effect on expression of Interleukin-6 (IL-6) and Tumor necrosis factor-α (TNF-α)from human renal glomerular endothelial cells ( HRGEC), to explore unkown pathogenesy of diabetic nephropathy.Methods Normal HRGEC was divided randomLy into normal glucose group(5 mmol/L D-glucose),manicol group (5 mmol/L D-glucose+25 mmol/L manicol)and high glucose group (30 mmol/L D-glucose). Real-time PCR was used to detect IL-6 and TNF-α mRNA expression in each group, Euzymelinked Immunosorbent Assay (ELISA) was performed to examine the protein expression of IL-6 and TNF-α in supernatant after 24 hours' culture. HRGEC was then randomLy divided into two groups: single high glucose group and high glucose + MBL group. After 24 hours'culture with 30 mmol/L D-glucose, 30％ MBL deficiency human serum was added into two groups, 1μg/mL MBL was only added into high glucose + MBL group, continued the culturation for another 4 hours. Flow cytometry and Immunofluorescence technique were applied to evaluate MBL, C3 and Menbrane Attacks Complex(MAC) deposition on cell surface respectively. Real-time PCR and ELISA were performed to examine mRNA and protein expression of both IL-6 and TNF-α in each group. Results Compared with normal glucose group and manicol group, the mRNA and protein expression of IL-6 and TNF-α in high glucose group were increased (P ＜ 0. 05). Flow cytometry confirmed obvious MBL and C3 co-deposition and Immunofluorescence confirmed obvious MAC deposition on cell surface in high glucose+MBL group. Compared with single high glucose group, the mRNA and protein expression of IL-6 and TNF-a in high glucose+MBL group were significantly higher (P＜0. 05). Conclusion High glucose can bring inflammatory factors' overexpression from cultured HRGEC; high glucose together with MBL can bring MBL complement pathway activation and inflammatory factors' overexpression, this indicates that the activation of MBL complement pathway may be a potential unkown pathogenesy of diabetic nephropathy and its proinflammatory status.关键词
人肾小球内皮细胞/高糖/甘露聚糖结合凝集素/白细胞介素-6/肿瘤坏死因子-αKey words
Human renal glomerular endothelial cells/ High glucose/ Mannose binding lectin/ Interleukin-6/ Tumor Necrosis Factor-α引用本文复制引用
武晓慧,黄颂敏,樊文星,唐万欣,邱红渝..高糖和MBL途径的补体激活对人肾小球内皮细胞表达IL-6、TNF-α的影响[J].四川大学学报（医学版）,2011,42(1):90-94,5.基金项目
国家自然科学基金(批准号30771005)资助 （批准号30771005）
