Kędzierska Karolina, Sporniak-Tutak Katarzyna, Sindrewicz Krzysztof, Bober Joanna, Domański Leszek, Parafiniuk Mirosław, Urasińska Elżbieta, Ciechanowicz Andrzej, Domański Maciej, Smektała Tomasz, Masiuk Marek, Skrzypczak Wiesław, Ożgo Małgorzata, Kabat-Koperska Joanna, Ciechanowski Kazimierz
Department of Nephrology, Transplantology, and Internal Medicine, Pomeranian Medical University, Szczecin, Poland.
Department of Dental Surgery, Pomeranian Medical University, Szczecin, Poland.
Drug Des Devel Ther. 2014 Sep 30;8:1695-708. doi: 10.2147/DDDT.S64814. eCollection 2014.
The structural proteins of renal tubular epithelial cells may become a target for the toxic metabolites of immunosuppressants. These metabolites can modify the properties of the proteins, thereby affecting cell function, which is a possible explanation for the mechanism of immunosuppressive agents' toxicity. In our study, we evaluated the effect of two immunosuppressive strategies on protein expression in the kidneys of Wistar rats. Fragments of the rat kidneys were homogenized after cooling in liquid nitrogen and then dissolved in lysis buffer. The protein concentration in the samples was determined using a protein assay kit, and the proteins were separated by two-dimensional electrophoresis. The obtained gels were then stained with Coomassie Brilliant Blue, and their images were analyzed to evaluate differences in protein expression. Identification of selected proteins was then performed using mass spectrometry. We found that the immunosuppressive drugs used in popular regimens induce a series of changes in protein expression in target organs. The expression of proteins involved in drug, glucose, amino acid, and lipid metabolism was pronounced. However, to a lesser extent, we also observed changes in nuclear, structural, and transport proteins' synthesis. Very slight differences were observed between the group receiving cyclosporine, mycophenolate mofetil, and glucocorticoids (CMG) and the control group. In contrast, compared to the control group, animals receiving tacrolimus, mycophenolate mofetil, and glucocorticoids (TMG) exhibited higher expression of proteins responsible for renal drug metabolism and lower expression levels of cytoplasmic actin and the major urinary protein. In the TMG group, we observed higher expression of proteins responsible for drug metabolism and a decrease in the expression of respiratory chain enzymes (thioredoxin-2) and markers of distal renal tubular damage (heart fatty acid-binding protein) compared to expression in the CMG group. The consequences of the reported changes in protein expression require further study.
肾小管上皮细胞的结构蛋白可能成为免疫抑制剂毒性代谢产物的作用靶点。这些代谢产物可改变蛋白质的特性，从而影响细胞功能，这可能是免疫抑制剂毒性机制的一种解释。在我们的研究中，我们评估了两种免疫抑制策略对Wistar大鼠肾脏蛋白质表达的影响。大鼠肾脏组织在液氮中冷却后匀浆，然后溶解于裂解缓冲液中。使用蛋白质测定试剂盒测定样品中的蛋白质浓度，并通过二维电泳分离蛋白质。然后用考马斯亮蓝对所得凝胶进行染色，并分析其图像以评估蛋白质表达的差异。随后使用质谱法对选定的蛋白质进行鉴定。我们发现，常用方案中使用的免疫抑制药物会在靶器官中诱导一系列蛋白质表达变化。参与药物、葡萄糖、氨基酸和脂质代谢的蛋白质表达明显。然而，在较小程度上，我们也观察到核蛋白、结构蛋白和转运蛋白合成的变化。接受环孢素、霉酚酸酯和糖皮质激素（CMG）的组与对照组之间观察到非常细微的差异。相比之下，与对照组相比，接受他克莫司、霉酚酸酯和糖皮质激素（TMG）的动物表现出负责肾脏药物代谢的蛋白质表达较高，而细胞质肌动蛋白和主要尿蛋白的表达水平较低。在TMG组中，与CMG组相比，我们观察到负责药物代谢的蛋白质表达较高，呼吸链酶（硫氧还蛋白-2）和远端肾小管损伤标志物（心脏脂肪酸结合蛋白）的表达降低。所报道的蛋白质表达变化的后果需要进一步研究。
