陳肅霖;林瑞生;晉淑意營養科學研究所2026-03-249999-12-311999https://handle.ncl.edu.tw/11296/5rkfa7https://scholars.csmu.edu.tw/handle/123456789/6604本研究利用微小核法(MN)來探討檳榔塊成分(arecoline、catechin、glycyrrhizin和Cu2+)在中國倉鼠卵(CHO)細胞之細胞遺傳毒性並探討CuCl2分別與arecoline、catechin及glycyrrhizin間的交互遺傳毒性。並以MTT法探討此四種檳榔塊成分對CHO細胞存活率之影響。另外，以核分裂指數(NDI)和細胞型態觀察來探討此四種成分對CHO細胞之細胞毒性，亦以流體細胞測定法(flow cytometry)探討此四種成分影響CHO細胞之細胞凋亡及細胞週期中各期之分佈情形。 MTT之研究成果發現arecoline、catechin、glycyrrhizin和CuCl2均能抑制CHO細胞之存活率，而CuCl2和arecoline、catechin或glycyrrhizin共存時，CuCl2可分別有統計意義的降低此三種成分之細胞毒性。在細胞型態之觀察結果發現只有catechin會改變CHO細胞之細胞型態，但目前對於catechin會改變細胞型態的機制並不十分清楚，有待進一步探討。此外，此四種成分對CHO細胞之NDI的影響較複雜，arecoline和glycyrrhizin在低濃度加強NDI，高濃度則抑制；catechin和CuCl2則增加NDI。 遺傳毒性的研究結果顯示arecoline、catechin、glycyrrhizin及CuCl2均有統計意義的增加CHO細胞之微小核數，其中glycyrrhizin在0.1-3M之濃度範圍可劑量相關性地增加CHO卵巢細胞之微小核數。交互遺傳毒性之探討發現CuCl2可加強arecoline之遺傳毒性，對catechin則無此作用，但對glycyrrhizin則抑制其之遺傳毒性。這些結果顯示檳榔塊成分間之交互遺傳毒性相當複雜。 細胞凋亡之研究結果顯示，此四種檳榔塊成分均不造成細胞凋亡，亦不影響其細胞體積和細胞之顆粒度，但arecoline、catechin和glycyrrhizin似有抑制G0/G1 phase之作用。 結論是arecoline，catechin，glycyrrhizin和CuCl2單獨存在時對CHO細胞均有細胞毒性和遺傳毒性，但其共存時會呈現複雜的交互作用。Betel chewing is a widespread habit in Taiwan, Southeast Asia and India. According to the statistical data of 1994, we know that the amount of betel quid chewier is about 10-15% of total population on earth, and the consumers of betel quid are still growing. Unfortunately, many investigators have shown that the habit of chewing betel quid is correlated to the high incidence of oral cancer and esophageal carcinoma (Muir and Kirk,1960; Hirayama, 1986; IARC,1985). However, the chemical ingredients of betel quid are a complex mixture including arecoline, Cu2+, catechin and glycyrrhizin. Since betel quid chewing exposes the consumer to a mixture of these compounds, a study of cytogenotoxicity of these compounds are important to understand their beneficial and harmful effects. Therefore, in this study, micronucleus (MN) test was employed to measure the genotoxic potential of arecoline, catechin, glycyrrhizin and Cu2+ in Chinese hamster ovary (CHO) Cells. Our results showed that all of the four ingredients of betel quid, arecoline, catechin, glycyrrhizin and Cu2+, significantly elevated the number of micronucleated cells, and glycyrrhizin caused genotoxicity in a concentration-dependent manner. In addition, the genotoxic interactions of Cu2+ and arecoline, catechin or glycyrrhizin were complex. Cu2+ attenuated the genotoxicity of glycyrrhizin, however, potentiated the genotoxicity of arecoline, but did not affect the genotoxicity of catechin. In this study, MTT assay, nuclear division index (NDI) and cell morphological change were conducted to study the cytotoxicity of the four ingredients of betel quid in CHO cells. The results of MTT assay showed that all of the four compounds at high concentrations significantly decreased the viability of CHO cells, but Cu2+ attenuated the cytotoxicity of arecoline, catechin or glycyrrhizin, respectively. In observation of cell morphology found that only catechin could cause the morphological change in CHO cells. On the other hand, the results of NDI were complex, both arecoline and glycyrrhizin increased cell proliferation at lower concentrations, but depressed cell proliferation at higher concentrations. In addition, both catechin and CuCl2 increased cell proliferation. Finally, the results of flow cytometry showed that any of the four ingredients didn’t cause hypoploidy cells, change cellular size, and affect cellular glandularity. Obviously, the four ingredients didn’t cause apoptosis in CHO cells. According to these findings, it is concluded that all the four ingredients of betel quid cause both genotoxicity and cytotoxicity in CHO cells. However, the cytogenotoxic interactions among these ingredients of betel quid are complex.微小核法MTT法流式細胞儀分析thesis