【摘要】 目的 探讨1,25（OH）₂VD₃对血管紧张素Ⅱ（Ang II）诱导的足细胞内质网应激的影响，为临床上应用骨化三醇治疗足细胞损伤产生的蛋白尿提供理论依据。方法 体外培养小鼠足细胞系（MPC5）,随机分为对照组、Ang II组(10^-6mol/L的Ang II)、1,25（OH）₂VD₃组[10^-6mmol/L的Ang II+10^-8mol/L的1,25（OH）₂VD₃]。各组均在给药处理6、12、18、24 h后收集细胞，分别应用实时定量PCR、免疫荧光技术检测不同时间点各组样本中葡萄糖调节蛋白（GRP78）及蛋白激酶样内质网激酶（PERK）的mRNA表达量和蛋白表达情况。结果 与对照组比较，Ang II组GRP78、PERK mRNA表达量在各个时间点均显著增加（P<0.01）。与Ang II组比较，1,25（OH）₂VD₃组GRP78、PERK mRNA表达量在各个时间点均显著降低（P<0.01或<0.05）,相应的GRP78、PERK蛋白表达均较低。Ang II组给药处理12 h后GRP78、PERK mRNA表达量较6 h mRNA表达量均显著增加（P<0.01）;Ang II组给药处理18 h后GRP78、PERK mRNA表达量较12 h mRNA表达量均显著增加（P<0.01,P<0.05）。结论 AngⅡ可以诱导足细胞发生内质网应激，其发生强度与药物作用时间有关，而1,25（OH）₂VD₃能通过抑制AngⅡ诱导的足细胞内质网应激，起到保护足细胞的作用。更多还原

【Abstract】 Objective To investigate the mechanism of calcitriol on proteinuria induced by podocyte destruction by observing the correlation between the 1,25（OH）₂VD₃ and Ang II-induced endoplasmic reticulum stress in podocytes.Methods The mus musculus Podocyte cell line（MPC5） was cultured in vitro and randomly divided into three groups, namely, the blank control group（A）, the Ang II group(B, treated with 10^-6 mol/L angiotensin II), and the 1,25（OH）₂VD₃ combination group(C, treated with 10^-6 mol/L angiotensin II and 10^-8 mol/L 1,25（OH）₂VD₃). The cells of each group were collected after 6 h, 12 h, 18 h and 24 h of the treatment. The mRNA expression and protein expression of glucose regulated protein（GRP78） and protein kinase like endoplasmic reticulum kinase（PERK） in samples of each group at different time points were detected by real-time quantitative PCR and immunofluorescence techniques.Results Compared with those in group A, the expression levels of GRP78 and PERK mRNA in group B increased significantly at all time points（P<0.01）. Compared with those in group B, the expression levels of GRP78 and PERK mRNA in group C decreased significantly at all time points（P<0.01 or<0.05）, and the corresponding GRP78 and PERK protein expression levels were lower. The expression levels of GRP78 and PERK mRNA in the Ang II group were significantly increased at 12 h compared with those at 6 h（P<0.01）. The expression levels of GRP78 PERK mRNA in the Ang II group increased significantly at 18 h compared with those at 12 h（P<0.01 or<0.05）, and the expression levels of GRP78 and PERK mRNA increased significantly at 18 h compared with 12 h（P<0.01 or<0.05）.Conclusion Ang II can induce endoplasmic reticulum stress in podocytes and the strength of ERs is related to the administration time. However, 1,25（OH）₂VD₃ can protect podocytes by inhibiting Ang II-induced ERs in podocytes.更多还原