Abstract

Cyclophosphamide (CP) is one of the most extensively used antineoplastic drug, but the nephrotoxicity caused by this drug is a major limiting factor for its use. Nerolidol (NERO) is a natural bioactive compound with diverse pharmacological actions. In Vitro and in vivo study was performed using HK-2 renal cells and Swiss Albino mice. Cell lines and animals were treated with NERO 25 and 50 µM + 30 µM CP (in vitro), 200 and 400 mg/kg, p.o. NERO from day 1 to day 15 + 200 mg/kg, i.p. CP on day 17 as single intraperitoneal injection (in vivo). The makers of oxidative stress, renal-specific injury markers, inflammation, apoptosis, fibrosis, and histopathological changes were studied. The study’s outcome showed a significant reduction in the level of malonaldehyde and interleukin-6 (p < 0.01), tumor necrosis factor-α, IL-1β (p < 0.001), and an increase in the superoxide dismutase, catalase, glutathione and interleukin-10 level (p < 0.01), in the in vivo study when treated with NERO 400 and compared with CP 200. In Vitro study showed reduced expression of nuclear factor kappa light chain enhancer of activated B cells, cleaved caspase-3, kidney injury molecule-1 and transforming growth factor-β-1 (p < 0.001), when treated with NERO 50 µM whereas NERO 25 µM only reduced the level of cleaved caspase-3 (p < 0.05) when compared with 30 µM. NERO 400 also reduced uric acid (p < 0.05), urea (p < 0.01), blood urea nitrogen, and serum creatinine levels (p < 0.001) and increased the level of blood-urea-nitrogen/creatinine ratio (p < 0.001). Additionally, the level of fibrosis-specific markers such as transforming growth factor-β1, hyaluronic acid (p < 0.01), 4-hydroxyproline, a collagen-rich area in Masson’s’ trichome stain, and Smad3 expression was also significantly reduced (p < 0.001). Furthermore, the outcome of multiple renal staining showed structural reversal aberrations, reduction of the thick basement membrane, and glycogen level toward normal when treated with NERO 400. Thus, the study showed a novel mechanistic modality of NERO against cyclophosphamide-induced renal toxicity. The outcome of this study can be considered a step closer to the development of an adjuvant to mitigate cyclophosphamide-induced renal toxicity among patients treated with cyclophosphamide.

摘要

环磷酰胺（CP）是最广泛使用的抗肿瘤药物之一，但该药物引起的肾毒性是其使用的主要限制因素。橙花叔醇 (NERO) 是一种具有多种药理作用的天然生物活性化合物。使用 HK-2 肾细胞和瑞士白化小鼠进行体外和体内研究。细胞系和动物用 NERO 25 和 50 µM + 30 µM CP（体外）、200 和 400 mg/kg，第 1 天至第 15 天口服 NERO + 200 mg/kg，第 17 天腹膜内注射 CP 进行单次腹膜内注射注射（体内）。研究了氧化应激、肾脏特异性损伤标志物、炎症、细胞凋亡、纤维化和组织病理学变化的标志物。研究结果显示，丙二醛和白细胞介素 6 (p < 0.01)、肿瘤坏死因子-α、IL-1β (p < 0.001) 水平显着降低，超氧化物歧化酶、过氧化氢酶、谷胱甘肽和白细胞介素水平增加-10 水平（p < 0.01），在体内研究中，用 NERO 400 处理并与 CP 200 相比。体外研究显示活化 B 细胞的核因子 kappa 轻链增强子、裂解的 caspase-3、肾损伤的表达降低当用 NERO 50 µM 处理时，分子-1 和转化生长因子-β-1 (p < 0.001)，而与 30 µM 相比，NERO 25 µM 仅降低了裂解 caspase-3 的水平 (p < 0.05)。NERO 400 还降低尿酸 (p < 0.05)、尿素 (p < 0.01)、血尿素氮和血清肌酐水平 (p < 0.001)，并增加血液尿素氮/肌酐比值 (p < 0.001) 。此外，纤维化特异性标志物的水平，如转化生长因子-β1、透明质酸 (p < 0.01)、4-羟脯氨酸、Masson 毛状体染色中富含胶原蛋白的区域以及 Smad3 表达也显着降低 (p < 0.01)。 0.001）。此外，用 NERO 400 治疗时，多重肾脏染色的结果显示结构逆转畸变、厚基底膜减少以及糖原水平趋向正常。因此，该研究显示了 NERO 对抗环磷酰胺诱导的肾毒性的新机制模式。这项研究的结果可以被认为是距离开发一种佐剂以减轻环磷酰胺治疗患者中环磷酰胺引起的肾毒性又近了一步。