陰道鞭毛蟲 (Trichomonas vaginalis, 30236 (JH31A, USA))主要寄生於人類的泌尿生殖道，是一種厭氧性的單細胞原生寄生蟲，是目前最常見的非病毒性的性接觸傳染疾病，受到感染的女性會引起陰道鞭毛蟲感染症 (trichomoniasis)。子宮頸癌是常見的婦女癌症之一，有研究調查指出陰道鞭毛蟲感染後會增加女性子宮頸癌的發生率，故推測女性子宮頸癌的生成與陰道鞭毛蟲感染症的細胞反覆發炎有關。研究也有指出感染陰道鞭毛蟲後的病患受到人類後天免疫缺乏病毒(HIV-1)感染的機會亦提高1.5倍。先前已有部分研究將陰道鞭毛蟲與不同的宿主細胞(fibroblast、MDCK…等)進行共同培養並且觀察，顯示陰道鞭毛蟲會黏附在不同的細胞表面，並以接觸性毒殺作用導致細胞脫落、受傷或死亡，但是對於研究陰道鞭毛蟲與子宮頸癌的發生並不是一個很好的模式。本實驗主要是想透過共同培養的模式觀察陰道鞭毛蟲是否會對人類子宮頸上皮細胞造成病變。先將陰道鞭毛蟲與子宮頸上皮細胞(Z172)馴化至相同環境培養液後進行共同培養；並以曠時攝影的方式觀察到隨著時間的增加，Z172細胞被陰道鞭毛蟲攻擊後開始內縮、細胞變圓，最後整片脫落等現象。進一步將影像細部分析統計，發現受到攻擊的細胞發生細胞脫落佔70％、類細胞凋亡約佔8％、類細胞壞死佔18％，以及RT-PCR的結果顯示，隨著共同培養的時間增加，陰道鞭毛蟲的黏附因子相關基因AP65、AP65-1與紅素氧還蛋白(rubrerythrin)都有表現增加的情形。而在人類子宮頸上皮細胞（Z172）的變化則是利用Oligo GEArray® Human Common Cytokines Superoarray 進行分析，發現隨著共同培養的時間增加，細胞中與細胞激素相關的基因如：BMP2、GDF 15、IL-8、IL-1α、IL-1β、ATP6AP1、VEGFB有增加的趨勢，從Cytokines Superoarray的量化結果中可得知BMP2上升1.75倍、GDF 15上升2.65倍、IL-8上升1.53倍、IL-1α上升3.25倍、IL-1β上升1.40倍、ATP6AP1上升1. 50倍、VEGFB上升1.63倍，而IL-8、IL-1α、IL-1β這些基因在炎症與免疫反應中均扮演著重要的角色，對於寄生蟲與宿主間的關係有很大的影響，而這些基因的上升在其他寄生蟲中也有相似的表現。由實驗結果可知，共同培養的時間增加，細胞型態的改變是透過陰道鞭毛蟲的黏附也就是接觸性毒殺作用所造成細胞病變，同時也可能以透過細胞激素的表現改變進而使得細胞受到傷害，但是對於陰道鞭毛蟲是如何導致子宮頸癌的發生，詳細的機制未來還需要做更進一步的研究與探討。

Trichomonas vaginalis (ATCC 30236 (JH31A)), a protozoan parasite of the urogenital-vaginal tract, is the causative agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in human. Although asymptomatic infection by T. vaginalis is common, multiple symptoms and pathologies can arise in women, including vaginitis, urethritis, prostatitis, low-birth weight infants and preterm delivery that cause of infertility. T. vaginalis infection was found to contribute the risk of cervical cancer, as determined by crude estimates and after adjustment for potential confounding effects. The attendant clump of this protozoan will destruct the epithelial cell and induce pathogenesis by contact-dependent cytotoxicty. In this study, T. vaginalis and human cervical epithelium cell (Z172) were raised under the same environment and with same culture medium. During T. vaginalis attached Z172, cell became round shape then shrinkage, part of cells dead, and disruption after co-cultured 10 hours respectively. Time-lapse recording and flow cytometry were used for the studies of the host and parasite interaction. Z172 interaction with T. vaginalis, there are 70% of cell with disruption, 8% with apoptosis-like and 18% with necrosis-like morphology after 10 hours. RT-PCR results showed that the expression of adhesion protein 65 of protozoa was increased after the interaction of host and parasite. Cell cytotoxicity was decreased after 6 hours interaction and increased 10 hours, and the expression of rubrerythrin in T. vaginalis was the same pattern as the host cell. In human genes by using Oligo GEArray® Human Common Cytokines SuperArray, we found that seven of inflammatory genes were unregulated in contrast with control such as BMP2 and GDF15 and IL-8 and IL-1α and IL-1β and ATP6AP1 and VEGFB；we found the genes were up 1.75 times of BMP2, 2.65 times of GDF 15, 1.53 times of IL-8, 3.25 times of IL-1α, 1.40 times of IL-1β, 1.50 times of ATP6AP1 and 1.63 times of VEGFB. Finally, in this study the results of experiments showed that contact-dependent cytotoxicity of Z172 by T. vaginalis. The cytopathogenic effect was increases along with the time during Z172 attacked by T. vaginalis and morphological mainly is disruption. The contact-dependent cytotoxicty change of the human cervical cell derived by the mechanical or chemical effect by the protozoan will be studied in the future.

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