Qu Liang, Qu Feng, Jia Zhixin, Wang Caihong, Wu Caisheng, Zhang Jinlan
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
J Ethnopharmacol. 2015 Jul 21;170:28-38. doi: 10.1016/j.jep.2015.05.010. Epub 2015 May 13.
Tripterygium wilfordii Hook F (TWHF) is a traditional herbal medicine in China. Triptolide (TP), the primary bioactive compound of TWHF, is an anti-inflammatory and immunosuppressive compound that can also injure the liver and kidney. Unfortunately, the toxicity mechanism remains unknown.
The aim of this study is to understand the regulatory role of sphingolipid (SPL) pathways in the TP-induced toxic mechanism in the liver and kidney in delayed-type hypersensitivity (DTH) Balb\c mouse.
76 core sphingolipids and 29 species of related metabolic enzymes in liver, kidney and plasma were analyzed with previous HPLC-MS/MS and real time qPCR method, respectively. Furthermore, the data generated from these two omics underwent integrated analysis to describe TP-induced abnormal sphingolipid metabolism and identify the specific biomarkers of TP toxicity using bioinformation method.
High-dose (LD50) TP could induce severe liver and kidney injuries. Moreover, TP comprehensively influenced the enzymes involved in the sphingolipids metabolism in the liver and kidney at the mRNA expression level. Furthermore, the total levels of ceramides (Cers), sphingomyelins (SMs) and sphingosine (Sph) were all elevated, while dihydroceramides (dhCers) and hexosylceramides (HexCers) were all down-regulated. Several enzymes, including kdsr, CerS2, CerS4, CerS5 and CerS6 in the liver and Cerk in the kidney were probably responsible for the TP-induced toxic effect, identifying them as possible novel therapeutic targets. Besides, fractions of long chain SPL (C16-C20) exhibited significant increase, and fractions of unsaturated dhCer and Cer were significantly changed, both of which above may be due to the change of mRNA expression level of CerSs. Moreover, several biomarkers for the diagnosis of TP poisoning were discovered.
In summary, the regulation of SPL metabolism uncovered a novel mechanism underlying TP poisoning in the liver and kidney. In addition, key biomarkers and enzymes may play an important role in reducing the clinical risk associated with the use of TP.
雷公藤（Tripterygium wilfordii Hook F，TWHF）是中国的一种传统草药。雷公藤甲素（TP）是TWHF的主要生物活性化合物，是一种具有抗炎和免疫抑制作用的化合物，但也会损伤肝脏和肾脏。不幸的是，其毒性机制尚不清楚。
本研究旨在了解鞘脂（SPL）途径在迟发型超敏反应（DTH）Balb\c小鼠肝脏和肾脏中TP诱导的毒性机制中的调节作用。
分别采用先前的高效液相色谱-串联质谱法（HPLC-MS/MS）和实时定量聚合酶链反应（qPCR）方法分析肝脏、肾脏和血浆中的76种核心鞘脂和29种相关代谢酶。此外，对这两种组学产生的数据进行综合分析，以描述TP诱导的异常鞘脂代谢，并使用生物信息学方法鉴定TP毒性的特定生物标志物。
高剂量（LD50）TP可导致严重的肝脏和肾脏损伤。此外，TP在mRNA表达水平上全面影响肝脏和肾脏中参与鞘脂代谢的酶。此外，神经酰胺（Cers）、鞘磷脂（SMs）和鞘氨醇（Sph）的总水平均升高，而二氢神经酰胺（dhCers）和己糖神经酰胺（HexCers）均下调。肝脏中的几种酶，包括kdsr、CerS2、CerS4、CerS5和CerS6以及肾脏中的Cerk可能是TP诱导毒性作用的原因，将它们确定为可能的新型治疗靶点。此外，长链SPL（C16-C20）组分显著增加，不饱和dhCer和Cer组分显著变化，上述两者可能均归因于CerSs mRNA表达水平的变化。此外，发现了几种用于诊断TP中毒的生物标志物。
总之，SPL代谢的调节揭示了TP导致肝脏和肾脏中毒的新机制。此外，关键生物标志物和酶可能在降低与使用TP相关的临床风险方面发挥重要作用。
