基于JAK2-STAT3途径探讨黄连素对高糖诱导足细胞损伤的修复机制

Study on the repairment mechanism of berberine on podocyte injury induced by high glucose based on JAK2-STAT3 pathway

关键词:  糖尿病肾病  足细胞  黄连素  JAK2  STAT3

Key words:  diabetic nephropathy  podocytes  berberine  JAK2  STAT3

摘要:

[目的] 从JAK2-STAT3途径探讨黄连素对高糖诱导足细胞损伤的修复机制。[方法] 培养MPC-5条件永生化小鼠足细胞，将细胞分为正常糖浓度组（5 mmol/L葡萄糖）、高糖浓度组（30 mmol/L葡萄糖）、高糖+10 μmol/L黄连素组、高糖+25 μmol/L黄连素组和高糖+50 μmol/L黄连素组。正常糖浓度组常规培养；其余各组采用相应培养基培养48 h。噻唑蓝（MTT）法检测细胞存活率；Transwell实验检测细胞的迁移能力；白蛋白流量率检测法测定足细胞屏障功能；逆转录-聚合酶链反应（RT-PCR）和蛋白免疫印迹（Western Blot）法分别检测JAK2和STAT3 mRNA和蛋白表达。[结果] 与正常糖浓度组相比，高糖刺激后足细胞存活率明显降低，迁移能力提高，屏障功能下降，JAK2和STAT3 mRNA和蛋白表达量提高（均P<0.01）；黄连素各组能不同程度促进足细胞存活，降低迁移能力，提高其障功能，并降低JAK2和STAT3 mRNA和蛋白表达；且呈浓度依赖性（P<0.05，P<0.01）。[结论] 黄连素能保护高糖诱导的足细胞损伤，提高足细胞在高糖环境下的存活率、降低其迁移率，维持其屏障功能，其机制可能与调控JAK2和STAT3基因与蛋白表达有关。

Abstract:

[Objective] The mechanism of berberine on the repair of podocyte injury induced by high glucose was investigated from JAK2-STAT3 pathway.[Methods] MPC-5 immortalized mouse podocytes were cultured and divided into normal glucose concentration group (5 mmol/L glucose),high glucose concentration group (30 mmol/L glucose),high glucose + 10 μmol/L berberine Group,high glucose + 25 μmol/L berberine group and high sugar + 50 μmol/L berberine group. The normal sugar concentration group was routinely cultured;the other groups were cultured for 48 h in the corresponding medium. Cell viability was detected by MTT assay;cell migration ability was detected by transwell assay;podocyte barrier function was measured by albumin flow rate assay;JAK2 and STAT3 mRNA and protein expression were detected by RT-PCR and Western Blot assay,respectively.[Results] Compared with the normal glucose concentration group,the survival rate of podocytes was significantly decreased after high glucose stimulation,the migration ability was increased,the barrier function was decreased,and the mRNA and protein expression levels of JAK2 and STAT3 were increased (all P<0.01). It promoted the survival of podocytes,decreased the migration ability,increased the barrier function,and decreased the expression of JAK2 and STAT3 mRNA and protein and the concentration was dependently (P<0.05,P<0.01).[Conclusion] Berberine can protect podocyte injury induced by high glucose,improve the survival rate of podocytes in high glucose environment,reduce its migration rate and maintain its barrier function. The mechanism may be related to the regulation of JAK2 and STAT3 gene and protein expression.