Raphael L, Fish J C
Transplantation. 1987 May;43(5):703-8. doi: 10.1097/00007890-198705000-00020.
Renal damage caused by cyclosporine (CsA) has been documented. Clinical experiences have shown preservation injury further potentiates CsA nephrotoxicity. This study examined the mechanism of nephrotoxicity defined by changes in protein synthesis, DNA synthesis, and ornithine decarboxylase activity in an in vitro model. Initial results showed that CsA inhibited dog kidney epithelium cell (MDCK) replication at a dose of 200 ng after 24 hr (P less than .01) and 100 ng after 48 hr (P less than .01). Protein synthesis was inhibited with 100 ng after 24 and 48 hr (P less than .01). There was a reduction in ODC activity with 200 ng CsA (P less than .05). Methods for simulating transplant-related injuries were then developed. Under ischemic conditions, 18 hr were required before a synergistic effect with CsA produced a reduction in replication (P less than .05). Incubation of MDCK cells in preservative solution at 4 degrees C under hypoxic conditions resulted in a time-dependent reduction in synthetic and replicative capacity that plateaued at 24 hr (P less than .01). The next step was to simulate the clinical situation by combining treatments. MDCK cells were incubated for 24 hr in preservative solution under hypoxic conditions at 4 degrees C, and then CsA was added at defined intervals. The addition of CsA before 24 hr resulted in a significant decrease in cell replication (P less than .05) compared with CsA addition after 48 hr. Similar results were obtained with cells incubated for 48 hr in preservative solution with hypoxia. These data suggest that renal injury from ischemia and cold storage requires a period of cellular repair and replication. Administration of CsA before this period results in further renal injury. Our analysis offers an explanation of CsA nephrotoxicity seen in the human situation and, therefore, may provide a model for studying human nephrotoxicity.
环孢素（CsA）引起的肾损伤已有文献记载。临床经验表明，保存损伤会进一步增强CsA的肾毒性。本研究在体外模型中，通过蛋白质合成、DNA合成和鸟氨酸脱羧酶活性的变化来研究肾毒性的机制。初步结果显示，24小时后，200 ng剂量的CsA抑制犬肾上皮细胞（MDCK）复制（P<0.01），48小时后，100 ng剂量的CsA抑制其复制（P<0.01）。24小时和48小时后，100 ng剂量的CsA抑制蛋白质合成（P<0.01）。200 ng CsA使ODC活性降低（P<0.05）。随后开发了模拟移植相关损伤的方法。在缺血条件下，与CsA产生协同作用导致复制减少前需要18小时（P<0.05）。在缺氧条件下，将MDCK细胞在4℃的保存液中孵育，导致合成和复制能力随时间下降，在24小时达到平台期（P<0.01）。下一步是通过联合治疗模拟临床情况。将MDCK细胞在4℃缺氧条件下的保存液中孵育24小时，然后在规定时间间隔添加CsA。与48小时后添加CsA相比，24小时前添加CsA导致细胞复制显著减少（P<0.05）。在缺氧条件下于保存液中孵育48小时的细胞也得到了类似结果。这些数据表明，缺血和冷藏引起的肾损伤需要一段时间的细胞修复和复制。在此期间之前给予CsA会导致进一步的肾损伤。我们的分析解释了在人类情况下所见的CsA肾毒性，因此可能为研究人类肾毒性提供一个模型。
