| Summary: | 碩士 === 國防醫學院 === 生物及解剖學研究所 === 89 === In recent years, ingestion of mildewed sugarcane caused Huntington's disease (HD) like syndrome in Southern China. It prompted further investigation and the corresponding results showed that was due to the toxicity of 3-nitroprpionic acid (3-NP) produced by Arthrinum in mildewed sugarcane. Metabolic compromise with systemic 3-NP resulted in the degeneration of striatal cell, mimicking the pathology of HD. In vivo study had shown that estradiol （E2） was able to attenuate HD like syndrome, which induced by 3-NP. However, there was no direct evidence to indicate that E2 could prevent 3-NP-induced toxicity on the striatal cells. In this study aimed to test the protecting and/or therapeutic effect of E2 to 3-NP-induced toxicity on primary striatal cell culture. The primary striatal cell cultures were prepared from E14-17 embryos. The WST-1 test was examined for the viability of normal, E2 and 3-NP treated on the striatal cell culture. DAPI and GAD immunocyto-chemistry double stains were applied to reveal 3-NP toxicity. The data showed that cell viability of striatal cells were obviously increased when singly treated with 10-5M E2. It could be also discovered that according to the corresponding result of DAPI stain, apoptosis rate were depressed for cells treated with E2. Besides, the results of WST-1 assays and DAPI stain showed that if 3-NP was solely treated with dose of 3mM, toxicity response was detected after 6 hours and extended to the max degree after 48 hours. Furthermore, when pretreated with 10-5M E2 and then treated with 3mM 3-NP after 48 hours, cell viability was increased almost to the degree of control level and apoptosis rate was also apparently decreased. However, it could also be found in our experiment that if E2 was added after pretreated with 3mM 3-NP for 6 hours, the corresponding data of WST-1 assays would showed that the protection from E2 was still valid, while observable toxicity response was also detected form the DAPI stain of GABA neurons. These phenomena described above could be concluded that E2 will not protect neurons when 3-NP induced injury was activated on neurons. On the other hand, after concurrently treated with E2 and 3-NP for 6 hours, we could observe that 3-NP induced toxicity would be inhibited due to the protection of E2. Hence, E2 could protect striatal cells against 3-NP toxicity via various modulating mechanism.
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