Serkova N, Christians U, Flögel U, Pfeuffer J, Leibfritz D
Fachbereich Chemie/Biologie, Universität Bremen, Germany.
Chem Res Toxicol. 1997 Dec;10(12):1359-63. doi: 10.1021/tx970071k.
The toxic effect of the macrolide immunosuppressant sirolimus on cell metabolism of primary astrocytes was studied by multinuclear NMR spectroscopy of viable cells and perchloric acid (PCA) extracts and compared to the effects of the immunosuppressant cyclosporine. The addition of 5 mg/L sirolimus (5.5 mumol/L) induced swelling of primary astrocytes to 110% of the original volume. Alteration in astrocyte volume in the presence of sirolimus was accompanied by reduction of the following important cell osmolytes and amino acid metabolites: myo-inositol, -58 +/- 12% (mean +/- standard deviation, n = 5); taurine, -44 +/- 5%; glutamine, -13 +/- 2%; compared with control. Sirolimus altered glucose metabolism and partially inhibited the tricarboxylic acid (TCA) cycle: sigma TCA/sigma glycolyse = 1.36 +/- 0.09 (control, n = 3), 0.96 +/- 0.08 (with sirolimus). The increased concentration of phosphodiesters by sirolimus addition (glycerophosphoethanolamine, 52 +/- 18%; glycerophosphocholine, 61 +/- 14%; compared with control, n = 5) indicated disorders in phospholipid metabolism of cellular membranes. Addition of sirolimus led to a decline of the energy state in astrocytes: the concentration of phosphocreatine (PCr) decreased to 75% of control value within 60 min of perfusion with sirolimus and the nucleotide triphosphate (NTP) concentration to 85% within 90 min (n = 3). The effect of sirolimus on the cell metabolism of astrocytes equals that of the immunosuppressants cyclosporine and tacrolimus, the neurotoxicity of which is well-established in clinical studies. The results of this in vitro study indicate that sirolimus possesses neurotoxic potential as well.
通过对活细胞和高氯酸（PCA）提取物进行多核核磁共振光谱研究，探讨了大环内酯类免疫抑制剂西罗莫司对原代星形胶质细胞代谢的毒性作用，并与免疫抑制剂环孢素的作用进行了比较。添加5mg/L西罗莫司（5.5μmol/L）可使原代星形胶质细胞肿胀至原始体积的110%。西罗莫司存在时星形胶质细胞体积的改变伴随着以下重要细胞渗透溶质和氨基酸代谢产物的减少：肌醇，-58±12%（平均值±标准差，n = 5）；牛磺酸，-44±5%；谷氨酰胺，-13±2%；与对照组相比。西罗莫司改变了葡萄糖代谢并部分抑制了三羧酸（TCA）循环：σTCA/σ糖酵解 = 1.36±0.09（对照组，n = 3），0.96±0.08（添加西罗莫司）。添加西罗莫司导致磷酸二酯浓度增加（甘油磷酸乙醇胺，52±18%；甘油磷酸胆碱，61±14%；与对照组相比，n = 5），表明细胞膜磷脂代谢紊乱。添加西罗莫司导致星形胶质细胞能量状态下降：在用西罗莫司灌注60分钟内，磷酸肌酸（PCr）浓度降至对照值的75%，在90分钟内三磷酸核苷酸（NTP）浓度降至85%（n = 3）。西罗莫司对星形胶质细胞代谢的影响与免疫抑制剂环孢素和他克莫司相当，后两者的神经毒性在临床研究中已得到充分证实。这项体外研究结果表明，西罗莫司也具有神经毒性潜力。
