title: Triptolide disrupts fatty acids and peroxisome proliferator-activated receptor (PPAR) levels in male mice testes followed by testicular injury: A GC-MS based metabolomics study.
abstract: Triptolide is the major active ingredient of Tripterygium Glycosides (TG), a traditional Chinese medicine with very potent anti-inflammatory effects and has been used in China for the treatment of rheumatoid arthritis and many other inflammatory diseases. However, clinical application of triptolide is restricted due to its multiple side effects, especially male infertility. The mechanism of triptolide on reproduction toxicity remains unclear. In the present study, a GC-MS based metabolomic approach was employed to evaluate the mechanism of triptolide-induced reproductive toxicity as well as identify potential novel biomarkers for the early detection of spermatogenesis dysfunction. In brief, male mice were divided into two groups with or without triptolide intraperitoneal injection at 60 μg/kg/day for 2 weeks and toxic effect of triptolide on testicular tissues were examined by biochemical indicator analysis, testis histopathologic analysis, and sperm quantity analysis. Metabolomics technology was then performed to evaluate systematically the endogenous metabolites profiling. Our results demonstrated that triptolide suppressed the marker-enzymes of spermatogenesis and testosterone levels, decreased sperm counts, reduced the gonad index and destroyed the microstructure of testis. Multivariate data analysis revealed that mice with triptolide induced testicular toxicity could be distinctively differentiated from normal animals and 35 and 39 small molecule metabolites were changed significantly in testis and serum, respectively (Fold-changes >1.5, P<0.05), in triptolide-treated mice. Abnormal level of fatty acids, an important energy source of sertoli cells with critical role in maintaining normal function of the testis tissue, was observed in triptolide-treated mice. Additionally, the protein expressions of PPAR, a transcription factor known to play a pivotal role in lipid and energy metabolism was significantly decreased in the testis tissue of triptolide-treated mice. In summary, our study represents the first comprehensive GC-MS based metabolomics analysis of triptolide-induced testicular toxicity. We reported for the first time that exposure to triptolide led to marked changes of a panel of endogenous metabolites in both testis and serum. The impairment of spermatogenesis may be caused by abnormal lipid and energy metabolism in testis via the down-regulation of PPARs mediated by triptolide. The presence of research suggested that PPARs and its related fatty acids metabolism may serve as potential targets for intervention or treatment of male infertility induced by triptolide.
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Ma
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Qi
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Chi
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Zhu
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China.
authors.forename: Lisha
authors.initials: L
authors.lastname: Yu
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, University of Iowa, Iowa City, IA, USA.
authors.forename: Guohua
authors.initials: G
authors.lastname: An
authors.affiliation: School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, People's Republic of China. Electronic address: nancyzhang03@hotmail.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000108:Acetylcarnitine / Q000032:analysis; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000633:toxicity*; D002331:Carnitine / Q000032:analysis; D004224:Diterpenes / Q000633:toxicity*; D004852:Epoxy Compounds / Q000633:toxicity; D005227:Fatty Acids / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry; D008297:Male; D055432:Metabolomics; D051379:Mice; D047492:Peroxisome Proliferator-Activated Receptors / Q000032:analysis*; D010616:Phenanthrenes / Q000633:toxicity*; D013091:Spermatogenesis / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects; D013737:Testis / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D013739:Testosterone / Q000097:blood
chemical_list: D000893:Anti-Inflammatory Agents; D004224:Diterpenes; D004852:Epoxy Compounds; D005227:Fatty Acids; D047492:Peroxisome Proliferator-Activated Receptors; D010616:Phenanthrenes; C001899:triptolide; D013739:Testosterone; D000108:Acetylcarnitine; D002331:Carnitine
keywords: GC–MS; Metabolomics; PPARs; Spermatozoa; Testicular toxicity; Triptolide