目的观察小檗碱对高糖培养足细胞黏附功能的影响,并探讨其改善高糖所致足细胞损伤的作用途径和分子机制。方法采用温度选择法体外进行永生化小鼠足细胞株的分化培养、诱导成熟,分为正常糖对照组(5 mmol/L葡萄糖)、高糖模型组(30 mmol/L葡萄糖)、甘露醇高渗对照组(30 mmol/L甘露醇)、高糖+小檗碱治疗组(30 mmol/L葡萄糖+1.25μmol/L小檗碱)。干预24 h、48 h、72 h后收集细胞,采用MTT法检测足细胞活力,离心法检测细胞黏附力,并采用Q-PCR法检测黏附分子α3、β1的mRNA表达水平。结果与正常糖对照组和甘露醇高渗对照组比较,高糖模型组在3个时间点的足细胞活力、黏附力和α3、β1的mRNA表达水平都明显下降(P<0.01),且随着作用时间的延长足细胞活力和黏附力降低越明显,黏附分子α3、β1的mRNA表达水平越低下(P<0.05);与高糖模型组比较,高糖+小檗碱治疗组的足细胞活力和黏附力明显增强(P<0.01),黏附分子α3、β1的mRNA表达水平明显升高,差异有统计学意义(P<0.01)。结论高浓度葡萄糖环境可降低足细胞活力和黏附力,改变足细胞黏附分子的mRNA表达水平;小檗碱可抑制和减轻高浓度葡萄糖对足细胞的损伤作用,可提升高浓度葡萄糖培养的足细胞的细胞活力和黏附力,对高浓度葡萄糖培养的足细胞具有明显的保护作用。
Objective To observe the effect of berberine on adhesion of podocytes incubated with high glucose and to explore the action and molecular mechanism of the compound in improving podocyte injury induced by high glucose. Methods Differentiation culture and induced maturation of immortalized mouse podocyte cell line was performed in vitro by temperature-selective method. The cells were divided into normal control group(5 mmol/L glucose), model group(30 mmol/L glucose), hypertonic control group(30 mmol/L mannitol) and treatment group(30 mmol/L glucose + 1.25 μmol/L berberine). Cells were collected after treatment for 24 h, 48 h and 72 h. Cell viability was detected by MTT assay, cell adhesion was determined by centrifugation and the mRNA expression of adhesion molecules α3 and β1 was detected by Q-PCR. Results Compared with the normal control group and the hypertonic control group, viability and adhesion of podocytes and mRNA expression of α3 and β1 were significantly reduced in a time-dependent manner at 3 time points(P<0.01), and were reduced further over time(P<0.05) in the model group. Compared with the model group, viability and adhesion of podocytes and mRNA expression of α3 and β1 were significantly increased in the treatment group(P<0.01). Conclusion High glucose environment reduces viability and adhesion of podocytes, and changes the mRNA expression of adhesion molecules in these cells. Berberine inhibits and reduces the damage of podocytes induced by glucose at high concentration, enhances viability and adhesion of podocytes incubated with high glucose, and has remarkable protective effect on these podocytes.
Shanghai Journal of Traditional Chinese Medicine