真武汤调节鞘氨醇激酶1-磷酸鞘氨醇信号通路抗阿霉素肾病大鼠肾损伤的作用研究

Efficacy of the Zhenwu decoction on regulating SphK1-S1P signaling pathway against renal injury in rats with adriamycin nephropathy

目的:研究真武汤对阿霉素肾病(Adriamycin Nephropathy,AN)模型大鼠鞘氨醇激酶1(Sphingosine Kinase 1,SphK1)-磷酸鞘氨醇(Sphingosine-1-phosphate,S1P)信号分子的影响,探讨真武汤治疗阿霉素肾病大鼠蛋白尿的机制。方法:常规检测各试验组动物血脂、血浆白蛋白、24小时尿蛋白含量;实时聚合酶链反应(Real-time Polymerase Chain Reaction,RTPCR)检测肾组织中SphK1基因表达;免疫印迹法(Western Blot)检测肾组织中SphK1蛋白表达;液相色谱-串联质谱法(Liquid Chromatography-tandem Mass Spectrometry,LC-MS/MS)检测SphK1、S1P活性。结果:模型组大鼠血脂水平、24小时尿蛋白含量显著升高,血浆白蛋白水平显著降低(P<0.05),肾脏SphK1酶活性、蛋白表达及S1P含量显著升高(P<0.05);与模型组相比,各给药组血脂水平、24小时尿蛋白含量明显降低,血浆白蛋白水平明显升高(P<0.05),肾脏SphK1酶活性、基因和蛋白表达及S1P含量明显下降(P<0.05),各治疗组之间差异无统计学意义。结论:真武汤可减少AN模型大鼠肾组织的SphK1酶活性、蛋白表达及S1P含量,改善其蛋白尿、血脂及血浆白蛋白水平。其作用可能与抑制SphK1、S1P信号分子的表达有关。

Objective:To study the effect of the Zhenwu decoction(真武汤)on the SphK1-S1P pathway and on the treatment of proteinuria in adriamycin nephropathy(AN)rats.Methods:Serum lipids,albumin and 24-hour urine protein were detected by routine methods.RT-PCR and Western blot were used to observe the expression of SphK1.LC-MC/MC was used to detect the activity of SphK1and S1P.Results:The levels of serum lipid,24-hour urine protein,the renal SphK1 enzyme activity,gene and protein expression and S1P in model group were significantly increased(P<0.05),whereas the level of albumin was obviously decreased(P<0.05).Interestingly,the increased level of serum lipid,24-hour urine protein,the renal SphK1 enzyme activity,gene and protein expression and S1P in all treatment groups were significantly decreased,whereas the level of albumin was dramatically increased in all medicine groups(P<0.05).Conclusion:The Zhenwu decoction can decrease the levels of proteinuria,SphK1 enzyme activity,gene and protein express in kidney tissue of AN rats,and improve the levels of blood lipid and albumin in AN rats.The effect of the Zhenwu decoction on treating albuminuria in AN rats may be related to the expression of SphK1 and S1P.