Department of Physiology, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, 210009, China.
School of Pharmacy, China Pharmaceutical University, Nanjing, 211198, China.
Chem Biol Interact. 2021 Nov 1;349:109662. doi: 10.1016/j.cbi.2021.109662. Epub 2021 Sep 21.
As a widely used anticancer drug in the clinic, cisplatin has obvious side effects, especially nephrotoxicity. Previous studies have suggested that the accumulation of intracellular reactive oxygen species (ROS) is a hallmark of cisplatin-induced acute kidney injury. This study aimed to investigate the relationship between ROS accumulation induced by cisplatin and 5-HT degradation. In vivo, by HE and TUNEL staining, we found that cisplatin-induced renal lesions and apoptotic regions, which were located in proximal tubular epithelial cells, were also the regions in which tryptophan hydroxylase 1 (Tph1), aromatic l-amino acid decarboxylase (AADC), 5-HT receptor (5-HTR) and monoamine oxidase A (MAO-A) were overexpressed, as determined by immunohistochemistry. Notably, the 5-HTR antagonist sarpogrelate hydrochloride (SH) and the AADC inhibitor carbidopa (CDP) significantly attenuated cisplatin-induced increases in serum creatinine and blood urea nitrogen levels, renal ROS levels, oxidative stress (SOD activity and MDA), proinflammatory cytokine levels (NF-κB, TNF-α and IL-1β), proapoptotic factor levels (Bax, Bcl-2, C-caspase 3 and C-caspase 9) and the phosphorylation of p38 and STAT3, as well as renal lesions and apoptosis. The combination of SH and CDP could almost abolish the effects of cisplatin challenge. In vitro, the effects of cisplatin challenge and the inhibitory effects of SH and CDP were also observed in HK-2 cells. Additionally, similar to the combination of SH and CDP, the MAO-A inhibitor clorgyline could also abolish the effects of cisplatin challenge. More importantly, by western blotting, we detected that the upregulation of Tph1, AADC and MAO-A expression induced by cisplatin both in vivo and in vitro could be obviously suppressed by SH to decrease 5-HT synthesis and mitochondrial 5-HT degradation. Altogether, these findings suggested that cisplatin-induced nephrotoxicity is due to the activation of the 5-HT degradation system in proximal tubular epithelial cells, including 5-HTR and 5-HT synthesis and degradation. 5-HTR plays a role by mediating the expression of MAO-A and the 5-HT synthases Tph1 and AADC.
顺铂作为临床上广泛使用的抗癌药物，具有明显的副作用，尤其是肾毒性。先前的研究表明，细胞内活性氧（ROS）的积累是顺铂诱导急性肾损伤的标志。本研究旨在探讨顺铂诱导的 ROS 积累与 5-HT 降解之间的关系。在体内，通过 HE 和 TUNEL 染色，我们发现顺铂诱导的肾损伤和凋亡区域位于近端肾小管上皮细胞中，也是色氨酸羟化酶 1（Tph1）、芳香族 l-氨基酸脱羧酶（AADC）、5-HT 受体（5-HTR）和单胺氧化酶 A（MAO-A）过度表达的区域，这通过免疫组织化学检测得到了证实。值得注意的是，5-HTR 拮抗剂盐酸沙格雷酯（SH）和 AADC 抑制剂卡比多巴（CDP）显著减轻了顺铂引起的血清肌酐和血尿素氮水平、肾 ROS 水平、氧化应激（SOD 活性和 MDA）、促炎细胞因子水平（NF-κB、TNF-α 和 IL-1β）、促凋亡因子水平（Bax、Bcl-2、C-caspase 3 和 C-caspase 9）以及 p38 和 STAT3 的磷酸化水平，以及肾损伤和凋亡。SH 和 CDP 的联合使用几乎可以消除顺铂的作用。在体外，也观察到了顺铂的作用和 SH 和 CDP 的抑制作用在 HK-2 细胞中。此外，与 SH 和 CDP 的联合使用类似，MAO-A 抑制剂氯丙嗪也可以消除顺铂的作用。更重要的是，通过 Western blot，我们检测到顺铂在体内和体外均能明显上调 Tph1、AADC 和 MAO-A 的表达，而 SH 可明显抑制 5-HT 的合成和线粒体 5-HT 的降解，从而降低 5-HT 的合成。总之，这些发现表明，顺铂诱导的肾毒性是由于近端肾小管上皮细胞中 5-HT 降解系统的激活，包括 5-HTR 和 5-HT 的合成和降解。5-HTR 通过介导 MAO-A 和 5-HT 合成酶 Tph1 和 AADC 的表达发挥作用。
