Fluvastatin protects against puromycin aminonucleoside-induced podocyte injury by inhibiting TRPC6

氟伐他汀通过抑制TRPC6保护嘌呤霉素氨基核苷诱导的足细胞损伤

In the present study,we aimed to investigate the effect of puromycin aminonucleoside(PAN)on the expression and distribution of transient receptor potential canonical 6(TRPC6)of murine podocytes and further study the protective mechanism of fluvastatin on podocyte injury by TRPC6 in vitro.Podocytes were treated by PAN at different doses and at different time points.The expressions of TRPC6 at mRNA and protein levels were assessed.An immunofluorescent assay was used to observe the distribution of TRPC6.Cultured podocytes were then divided into four groups.The expressions of TRPC6 at mRNA and protein levels were measured.The intracellular calcium concentration of podocytes was measured with a laser-scanning confocal microscope.The paracellular permeability to BSA was evaluated using Millicell-PCF Inserts.The expressions of TRPC6 at mRNA and protein levels were increased in a dose and time-dependent manner after exposure to PAN.Immunofluorescence showed that the expression intensity of TRPC6 was significantly increased,and the distribution of TRPC6 was changed after PAN was applied to podocytes.With fluvastatin intervention,PAN-induced up-regulation of TRPC6 was significantly reversed.The ratio of the peak value of intracellular calcium to the basic calcium value in the PAN group was significantly higher after TRPC6 was activated by OAG,while it is obviously reversed under the action of fluvastatin.The podocyte permeability was significantly increased after 48 h of PAN treatment,while the above situation was effectively improved after fluvastatin intervention.The changes of distribution and expression of TRPC6 were related to podocyte injury induced by PAN.Fluvastatin could exert its protective effects on podocytes by down-regulating TRPC6.

在本研究中,我们观察了嘌呤霉素氨基核苷(PAN)对小鼠足细胞瞬时受体电位阳离子通道蛋白6(TRPC6)表达和分布的影响,并进一步研究了氟伐他汀对TRPC6所致足细胞损伤的保护机制。PAN在不同剂量和不同时间点处理足细胞,在mRNA和蛋白质水平上评估TRPC6的表达,免疫荧光法观察TRPC6的分布。然后再将体外培养的足细胞分为4组,分别检测TRPC6在mRNA和蛋白水平表达的变化,用激光共聚焦显微镜测定足细胞内钙浓度的变化,应用Millicell PCF评估BSA的细胞通透性。暴露于PAN后,TRPC6在mRNA和蛋白质水平的表达以剂量和时间依赖性方式增加。免疫荧光显示,将PAN应用于足细胞后,TRPC6的表达强度显著增加,并且TRPC6的分布发生改变。在氟伐他汀干预下,PAN诱导的TRPC6上调得以显著逆转。OAG激活TRPC6后,PAN组细胞内钙峰值与基础钙值的比值显著升高,而在氟伐他汀作用下则明显逆转。PAN处理48 h后足细胞通透性显著增加,而氟伐他汀干预后上述情况得到有效改善。TRPC6的分布和表达变化与PAN诱导足细胞损伤有关,氟伐他汀可通过下调TRPC6对足细胞发挥保护作用。