2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38624Pesticide exposure is associated with various neoplastic and degenerative diseases. Oxidative damage to mitochondrial DNA (mtDNA) has been also proposed to be an important mediator in apoptosis and carcinogenesis. Pesticides may be metabolized by hepatic cytochrome P450 3A5 enzymes (CYP3A5), or glutathione S-transferases. The human multidrug resistance 1 (MDR1) gene encodes a membrane transport protein P-glycoprotein, which plays an important role in protecting cells against pesticide toxicity by acting as an efflux pump to minimize intracellular exposure to pesticide. A structural variation in a nuclear-encoded mitochondrial complex I subunit gene NDUFV2, has been also observed to reduce the activity of respiratory complex I, and promoted the generation of reactive oxygen species (ROS) and free radicals that may cause the structural defects in the mitochondria. The manganese superoxide dismutase (MnSOD) is a defender against superoxide radicals; the 8-oxoguanine DNA glycosylase (OGG1) is also the major DNA glycosylase for the repair of 8-oxoG lesions in the mitochondrial DNA. However, the mitochondrial genotoxicity of pesticides in people with various genetic variations of human MnSOD and OGG1 have not been investigated. 8-hydroxy-2’-deoxyguanosine (8-OHdG) is one of the most abundant oxidative DNA damage, which has been used to determine the level of oxidative damage in individuals, and further to explore the relation with the mitochondrial DNA damage. As circulating soluble Fas (sFas) and soluble Fas ligand (sFasL) have been implicated in protection from Fas-mediated apoptosis, and may contribute the development of related diseases. Thus, this study will be designed to investigate whether various metabolic and transport protein, mitochondrial structure, antioxidative and DNA repair genotypes are more susceptible to mitochondrial DNA damage in pesticide-exposed fruit growers. We will also evaluate the relationships of cellular oxidative stress, mitochondrial DNA damage, and apoptosis elicited by pesticide exposure. The mitochondrial DNA damage in the peripheral blood of 450 fruit growers who experienced pesticide exposure and 180 unexposed controls will be quantified by real-time quantitative polymerase chain reaction (real time qPCR). Questionnaires will be also administered to obtain demographic data, cigarette smoking habits, medical, and occupational histories. Genotypes will be identified by polymerase chain reaction (PCR); urinary 8-OHdG level will be analyzed using liquid chromatography with tandem mass spectrometry (LC/MS/MS). Serum soluble Fas and FasL levels will be also detected with enzyme linked immunosorbent assay (ELISA) kits. Our anticipated result is that pesticide exposed individuals with susceptible metabolic CYP3A5 and GSTP1 genotypes, transport protein MDR1 genotype, mitochondrial structure NDUFV2 genotype, antioxidative MnSOD and DNA repair OGG1 genotypes are more likely to have a higher level of cellular oxidative stress, mitochondrial DNA damage, and antiapoptosis. Furthermore, the relationships of cellular oxidative stress, mitochondrial DNA damage, and apoptosis elicited by pesticide exposure will be observed. In summary, susceptible genes may modulate the mitochondrial DNA damage, cellular DNA oxidative damage, and apoptosis in pesticide-exposed fruit growers. Our study may shed more light on the development of disease caused by pesticide.農藥(pesticide) 暴露已經被觀察到是相關於若干的惡性腫瘤及退化性疾病；粒線體 DNA (mitochondrial DNA [mtDNA]) 的氧化傷害也已經被推測在細胞凋零(apoptosis) 與致癌機制上扮演重要的調節者。農藥可被肝臟細胞色素P450 3A5 (CYP3A5) 酵素或 是麩胺基硫轉移酶(glutathione S-transferases) 所代謝；細胞膜運送蛋白P-醣蛋白 (P-glycoprotein)，是人類多重抗藥性1 (multidrug resistance 1 [MDR1]) 基因的產物，可 藉由擔任輸出幫浦(efflux pump) 來減少細胞內之農藥暴露，因此扮演對抗農藥毒性以 保護細胞的重要角色。於細胞核轉碼粒線體複合體I (complex I) 次單元之基因NDUFV2 的結構變異，亦已經被觀察減低呼吸複合體I 之活性，促使反應性氧化物種(reactive oxygen species [ROS]) 與自由基(free radicals) 產生，即可能導致粒線體結構缺陷。含 錳超氧化物歧解酶(manganese superoxide dismutase [MnSOD]) 擔任對抗超氧自由基攻 擊的防禦者，8-oxoguanine DNA glycosylase (OGG1) 則也是主要的醣蛋白來修補粒線體 DNA 8-oxoG 受損部位；然而，具有MnSOD 和OGG1 基因變異者其在農藥所導致的粒 線體基因毒性並未被探討。8-hydroxy-2’-deoxyguanosine (8-OHdG) 是最為豐富的氧化性 DNA 傷害(oxidative DNA damage) 之一，已經被用以決定個體的氧化傷害程度，並加 以探討與粒線體DNA 損傷間的相關。而循環之可溶性Fas (sFas) 與可溶性Fas 配位基 (sFasL) 已經被認為可影響Fas 所調控的細胞凋零之保護作用，並且可能貢獻於相關疾 病之發展上。因此，本研究將被設計來探討在暴露於農藥之果農中，是否代謝與運送蛋 白基因、粒線體結構基因、抗氧化基因與DNA 修補基因對於其粒線體DNA 傷害較具 易感受性；我們也將瞭解農藥所引起的細胞氧化壓力、粒線體DNA 傷害、以及細胞凋 零之間的關係。我們將以即時定量聚合酶鏈鎖反應(real-time quantitative polymerase chain reaction [real time qPCR]) 來評估450 名經歷農藥暴露的果農與180 名未暴露的對 照其個人周邊血液的粒線體DNA 傷害程度；問卷也將被執行以獲取人口學資料、抽菸 習慣、疾病史以及職業史。基因型將是以聚合酶鏈鎖反應(polymerase chain reaction [PCR]) 加以判定， 尿液8-OHdG 濃度是利用液相層析儀串連質譜儀(liquid chromatography with tandem mass spectrometry [LC/MS/MS]) 所分析，血清可溶性Fas 及 FasL 濃度也將是使用酵素連結免疫吸附法(enzyme-linked immunosorbent assay [ELISA]) 來偵測。我們預期的結果是具有易感受性代謝CYP3A5 和GSTP1 基因型、運送蛋白 MDR1 基因型、粒線體結構NDUFV2 基因型、抗氧化MnSOD 基因型與DNA 修補OGG1 基因型之農藥暴露者是較可能具有細胞氧化壓力、粒線體DNA 傷害與抗細胞凋零的較 高表現，並且農藥所導致之細胞氧化壓力、粒線體DNA 傷害與抗細胞凋零表現之間的 相關性也將被觀察到。總體而言，易感受性基因可能調節農藥暴露之果農的粒線體DNA 傷害、細胞DNA 氧化傷害及細胞凋零效應；我們的研究可對於農藥的致病機制提供更 進一步的瞭解。農藥粒線體DNA傷害氧化性DNA傷害細胞凋零基因易感受性pesticidemitochondrial DNA damageoxidative DNA damageapoptosisgenetic susceptibilityProject