血栓素A2受体拮抗剂雷马曲班对足细胞凋亡和nephrin表达的影响

Effect of thromboxane A2 receptor antagonist ramatroban on podocyte apoptosis and nephrin protein expression

目的观察血栓素A2(TXA2)受体拮抗剂雷马曲班对嘌呤霉素氨基核苷(PAN)诱导的足细胞凋亡、nephrin mRNA和蛋白表达的影响。方法以条件永生性小鼠足细胞株为研究对象,随机分为4组:对照组、PAN组、雷马曲班组、PAN+雷马曲班组。PAN组加入100μg/mL PAN、雷马曲班组加入30nmol/L雷马曲班、PAN+雷马曲班组加入100μg/mL PAN和30nmol/L雷马曲班,3组均刺激48h。采用流式细胞仪检测足细胞凋亡水平,实时荧光定量RT-PCR检测nephrin mRNA的相对表达量,Western印迹法检测nephrin蛋白的表达状况。结果 PAN组的足细胞凋亡率显著高于对照组、雷马曲班组和PAN+雷马曲班组(P值均<0.05),nephrin mRNA和蛋白表达量显著低于对照组、雷马曲班组和PAN+雷马曲班组(P值均<0.05)。结论 TXA2受体拮抗剂雷马曲班可以抑制PAN诱导下的足细胞凋亡,使nephrin表达量上调。

Objective To investigate the effect of ramatroban （a thromboxane A2 receptor antagonist） on podocyte apoptosis induced by puromycin aminonucleoside （PAN） and expression of nephrin. Methods Conditionally immortalized murine podocytes were randomly divided into 4 groups; control group, PAN group, ramatroban group and PAN ＋ ramatroban group. Podocytes were exposed to 100 μg/mL PAN, 30 nmol/L ramatroban, and 100 μg/mL PAN combined with 30 nmol/L ramatroban for 48 h in the latter three groups, respectively. Then the apoptosis of podocytes was measured by flow cytometry. The expression of nephrin mRNA was detected by real-time quantitative reverse transcriptase polymerase chain reaction （RT-PCR）. The protein level of nephrin was detected by Western blotting. Results Compared with normal control group, ramatroban group and PAN＋ ramatroban group, the apoptosis of podocytes was significantly increased in PAN group （all P〈 0.05）, while mRNA and protein levels of nephrin were significantly decreased （all P〈0. 05）. Conclusion Ramatroban can inhibit the apoptosis of podocytes induced by PAN and up-regulate the protein expression of nephrin.