CAJ | 학술논문

目的用不同体外试验评价卷烟烟气粒相物提取液(CSCs)致人外周血淋巴细胞的细胞毒性和遗传毒性.方法分别以25、50、75、100和125 ug/ml的CSCs在加或不加肝脏微粒体酶(S9)系统下作用于人外周血淋巴细胞3 h,然后用CCK-8试验检测细胞毒性,用彗星试验检测DNA损伤,用hprt和TCR基因突变试验检测体细胞突变;以75ug/ml的CSCs作用于人外周血淋巴细胞3 h,分别给予30、60、90、120和240 min的修复时间,然后用彗星试验评价淋巴细胞的DNA修复情况.结果细胞活性随剂量增加明显降低,100、125 ug/ml CSCs加S9组细胞活性明显高于不加S9组,差异有统计学意义(P<0.05,P<0.01);随CSCs暴露剂量增加,DNA损伤明显增加,并明显高于对照组,差异有统计学意义(P<0.01);各剂量加S9组DNA损伤明显低于不加S9组,差异有统计学意义(P<0.05,P<0.01).各剂量组TCR基因突变率明显高于对照组,差异有统计学意义(P<0.05,P<0.01).中高剂量组hprt基因突变率明显高于对照,差异有统计学意义(P<0.01),中、高剂量加S9与不加S9两组间TCR和hprt基因突变率均存在明显差异,差异有统计学意义(P<0.05,P<0.01).加与不加S9两组DNA损伤均可基本修复至正常水平,但两者修复速度存在差异.结论 CSCs可在体外诱发人外周血淋巴细胞的细胞和遗传损伤,但S9可降低CSCs毒性的效应,并可影响人淋巴细胞DNA损伤的修复速率.
목적 용불동체외시험평개권연연기립상물제취액(CSCs)치인외주혈림파세포적세포독성화유전독성.방법 분별이25、50、75、100화125 ug/ml적CSCs재가혹불가간장미립체매(S9)계통하작용우인외주혈림파세포3 h,연후용CCK-8시험검측세포독성,용혜성시험검측DNA손상,용hprt화TCR기인돌변시험검측체세포돌변;이75ug/ml적CSCs작용우인외주혈림파세포3 h,분별급여30、60、90、120화240 min적수복시간,연후용혜성시험평개림파세포적DNA수복정황.결과 세포활성수제량증가명현강저,100、125 ug/ml CSCs가S9조세포활성명현고우불가S9조,차이유통계학의의(P<0.05,P<0.01);수CSCs폭로제량증가,DNA손상명현증가,병명현고우대조조,차이유통계학의의(P<0.01);각제량가S9조DNA손상명현저우불가S9조,차이유통계학의의(P<0.05,P<0.01).각제량조TCR기인돌변솔명현고우대조조,차이유통계학의의(P<0.05,P<0.01).중고제량조hprt기인돌변솔명현고우대조,차이유통계학의의(P<0.01),중、고제량가S9여불가S9량조간TCR화hprt기인돌변솔균존재명현차이,차이유통계학의의(P<0.05,P<0.01).가여불가S9량조DNA손상균가기본수복지정상수평,단량자수복속도존재차이.결론 CSCs가재체외유발인외주혈림파세포적세포화유전손상,단S9가강저CSCs독성적효응,병가영향인림파세포DNA손상적수복속솔.
Objective To investigate the cyto-genotoxicity of cigarette smoke condensates (CSCs) in human peripheral blood lymphocytes with different assays in vitro. Methods Human lymphocytes were ex-posed to particle matter of cigarette smoke combined with or without S9 mixtures at doses of 25,50,75,100 and 125 ug/ml for 3 h. The cytotoxicity induced by CSCs was detected by CCK-8 assay. The DNA damage, DNA repair (repair time: 30, 60, 90, 120 and 240 min, respectively) and the somatic cell mutations induced by 75 ug/ml CSCs were measured by comet assay, hprt gene and TCR gene mutation tests, respectively. Results CCK-8 assay indicated that the cell viability decreased with CSCs doses. At the doses of 100,125 ug/ml, the cell viability of CSCs +S9 group was significantly higher than that of CSCs -S9 group(P<0.05,P< 0.01). In comet assay, DNA damage significantly increased in a dose-dependent manner, as compared with controls (P<0.01). Moreover, there was significant difference between -S9 group and +S9 group (P<0.05, P< 0.01). The Mf-TCR at each dose group was significantly higher than that of controls (P<0.05 ,P<0.01). The Mf-hprt at high-dose groups were significantly higher than that of controls (P<0.01), and significant difference of Mf-TCR and Mf-hprt at high doses of CSCs between -S9 group and +S9 group (P<0.05,P<0.01). The DNA damage induced by CSCs+S9 or CSCs-S9 could be repaired, but DNA repair speed was different between -S9 group and +S9 group (P<0.05, P<0.01). Conclusion CSCs may induce cyto-genotoxicity in human periph-eral blood lymphocytes in vitro, but S9 mix could reduce the toxicity of CSCs and impact DNA repair speed.