Part I. Nephroprotective role of resveratrol and ursolic acid in aristolochic acid-treated zebrafishPart II. Effects of Topiramate on maternal oogenesis and offspring cartilage dysplasia

• Main Authors: Yu-Ju Ding, 丁玉如
• Other Authors: Yau-Hung Chen
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/22512323280759072272

博士 === 淡江大學 === 化學學系博士班 === 103 === Part I. Nephroprotective role of resveratrol and ursolic acid in aristolochic acid-treated zebrafish Aim of this study was to study the nephroprotective effects of resveratrol (Resv) and ursolic acid (UA) in a zebrafish model. Using two transgenic lines, Tg(wt1b:GFP) and Tg(gata1:dsRed) the subtle changes in the kidney and the red blood cells circulation can be easily recorded. Results showed that both Resv and UA treatment can attenuate AA-induced kidney injury and improve the blood circulation. We used glomerular filtration rate assays to evaluate zebrafish''s renal function and found that both Resv and UA treatment can restore renal function (100% for Mock; 56.1±17.3% for AA-treated; 80.2± 11.3% for Resv+AA; and 83.1± 8.1% for UA+AA, n=15). Furthermore, real time RT-PCR experiments show that pre-treatment with either Resv or UA suppresses pro-inflammatory gene expressions. NMR results revealed that tubular amino acid reabsorption can be improved by Resv or UA treated. In conclusion, our findings reveal that AA-induced nephrotoxicities can be attenuated by pre-treatment with either Resv or UA. Part II. Effects of Topiramate on maternal oogenesis and offspring cartilage dysplasia Topiramate is used to treat epilepsy in children and adults. In children, it is indicated for the treatment of Lennox-Gastaut syndrome, a disorder that causes seizures and developmental retardation. Recent clinical data showed that administration of Topiramate in pregnant women caused an increased risk of oral clefts on their new born babies. To further understand the adverse effects of Topiramate, we dosed adult female zebrafish with 0.5 mg/g/day for 7 days, and recorded the phenotypic changes of adult females as well as their offspring. In adults, histopathological examination showed that Topiramate treatment reduced oocyte maturation. Meanwhile, around 23.0% (n=14) of offspring derived from Topiramate-treated adults displayed cartilage deformity, such as ceratobranchial missing, ceratohyal, Meckel’s cartilage and ethmoid plate deformation. Alizarin red staining revealed that the mineralization of ceratohyal, Meckel’s cartilage, and vertebrae were downregulated. From the molecular points of view, real-time PCR demonstrated that the expression levels of smad 1/4/5 and runx2b were upregulated in the Topiramate-treated groups at different sites (ovaries, eggs and 1-cell-stage embryos) in comparison with those of mock-treated controls. Finally, whole-mount in situ hybridization on 8 hpf embryos indicated that endogenous runx2b levels were upregulated. Taken together, we concluded that Topiramate might induce maternal smad1/4/5 and runx2b genes overexpression, and then caused offspring skeletal dysplasia.