The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Center for Drug Safety Evaluation and Research, Innovation Research Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China.
Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, China.
Phytomedicine. 2019 May;58:152867. doi: 10.1016/j.phymed.2019.152867. Epub 2019 Feb 18.
Toosendan Fructus is traditionally used as an insecticide or digestive tract parasiticide for treating digestive parasites in China. It is recorded to have little toxicity in Chinese Pharmacopoeia and has been found to cause severe liver injury during clinical practice.
This study aims to identify candidate serum microRNAs (miRNAs) as potential toxicological biomarkers for reflecting the hepatotoxicity induced by toosendanin (TSN), which is the main toxic compound isolated from Toosendan Fructus METHODS: Alanine/aspartate aminotransferase (ALT/AST) activities detection and liver histological observation were performed to evaluate the liver injury induced by TSN or other hepatotoxicants in mice. miRNAs chip analysis and Real-time PCR assay were conducted to identify the altered miRNAs in serum from TSN-treated mice RESULTS: The results of serum ALT/AST and liver histological evaluation showed that TSN (10 mg/kg) induced hepatotoxicity in mice. The results of miRNAs chip showed that the expression of 81 serum miRNAs was obviously altered in mice treated with TSN for 12 h, and 22 of them have passed the further validation in serum from mice treated with TSN for both 6 h and 12 h. These 22 miRNAs were supposed to be the candidate toxicological biomarkers for TSN-induced hepatotoxicity with more sensitivity as compared to the alteration of AST or ALT activity. Moreover, the expression of miRNA-122-3p and mcmv-miRNA-m01-4-3p was not only increased in TSN-treated mice, but also increased in mice treated with other hepatotoxicants including acetaminophen (APAP), monocrotaline (MCT) and diosbuibin B (DB). Only the expression of serum miRNA-367-3p was increased in TSN-treated mice but not changed in the liver injury induced by APAP, MCT or DB CONCLUSION: miR-122-3p and mcmv-miRNA-m01-4-3p may be two commonly sensitive biomarkers for reflecting the hepatotoxicity induced by exogenous hepatotoxicants, and miR-367-3p may be a specific biomarker for reflecting the liver injury induced by TSN.
在中国，酸橙果实传统上被用作杀虫剂或消化道寄生虫药来治疗消化道寄生虫。它在《中国药典》中记录的毒性很小，但在临床实践中已发现它会导致严重的肝损伤。
本研究旨在鉴定候选血清 microRNAs（miRNAs）作为潜在的毒理学生物标志物，以反映从酸橙果实中分离出的主要毒性化合物川楝素（TSN）引起的肝毒性。
通过丙氨酸/天冬氨酸氨基转移酶（ALT/AST）活性检测和肝组织学观察来评估 TSN 或其他肝毒性剂在小鼠中引起的肝损伤。通过 miRNA 芯片分析和实时 PCR 检测来鉴定 TSN 处理小鼠血清中改变的 miRNAs。
血清 ALT/AST 和肝组织学评价结果表明，TSN（10mg/kg）在小鼠中诱导了肝毒性。miRNA 芯片结果显示，TSN 处理 12h 的小鼠血清中 81 种血清 miRNAs 的表达明显改变，其中 22 种 miRNA 已通过 TSN 处理 6h 和 12h 的小鼠血清进一步验证。与 AST 或 ALT 活性的改变相比，这些 22 种 miRNAs 被认为是 TSN 诱导肝毒性的候选毒理学生物标志物，具有更高的敏感性。此外，miRNA-122-3p 和 mcmv-miRNA-m01-4-3p 的表达不仅在 TSN 处理的小鼠中增加，而且在其他肝毒性剂（包括对乙酰氨基酚（APAP）、单环酸（MCT）和地布滨 B（DB））处理的小鼠中也增加。只有血清 miRNA-367-3p 的表达在 TSN 处理的小鼠中增加，而在 APAP、MCT 或 DB 诱导的肝损伤中没有改变。
miR-122-3p 和 mcmv-miRNA-m01-4-3p 可能是反映外源性肝毒性剂引起的肝毒性的两个共同敏感的生物标志物，而 miR-367-3p 可能是反映 TSN 引起的肝损伤的特异性生物标志物。
