CAJ | 학술논문

目的观察胚胎植入期二硫化碳(CS2)暴露对孕鼠血清雌二醇(E2)含量和子宫组织雌激素受体α(ER-α)表达水平的影响,探讨CS2致胚胎毒性的作用机制.方法健康雌性小鼠分别在受孕后第3天(GD3)、第4天(GD4)、第5天(GD5)、第6天(GD6)暴露于单一剂量(631.4 mg/kg)的CS2或橄榄油(溶剂对照),单次暴露之后在不同的观察终点结束实验.采用酶联免疫吸附试验(ELISA)检测各观察终点孕鼠血清E2含量,采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS)和蛋白免疫印迹法(Western blotting)检测各观察终点子宫组织ER-α表达水平.结果 GD3、GD4、GD5和GD6各暴露组在孕后第7天观察终点血清E2水平有不同程度的下降,差异均有统计学意义(P,＜0.05)；GD3和GD5暴露组孕鼠在孕后第6天观察终点其血清E2水平也有所下降,差异有统计学意义(P＜0.05).Western blotting结果显示,GD4和GD5暴露组孕鼠子宫组织中ER-α均在暴露后第一个观察终点(即孕后第5天和孕后第6天)低表达,表达水平分别较对照组降低了23.6％和72.9％,差异均具有统计学意义(P＜0.05)；GD3和GD6暴露组孕鼠在各观察终点ER-α表达水平均较对照组有所降低,但差异无统计学意义(P＞0.05).结论胚胎植入期CS2暴露可降低子宫组织ER-α表达水平并减少孕鼠血清E2含量,这可能是CS2致胚胎毒性的分子机制之一.
목적 관찰배태식입기이류화탄(CS2)폭로대잉서혈청자이순(E2)함량화자궁조직자격소수체α(ER-α)표체수평적영향,탐토CS2치배태독성적작용궤제.방법 건강자성소서분별재수잉후제3천(GD3)、제4천(GD4)、제5천(GD5)、제6천(GD6)폭로우단일제량(631.4 mg/kg)적CS2혹감람유(용제대조),단차폭로지후재불동적관찰종점결속실험.채용매련면역흡부시험(ELISA)검측각관찰종점잉서혈청E2함량,채용십이완기광산납-취병희선알응효전영(SDS)화단백면역인적법(Western blotting)검측각관찰종점자궁조직ER-α표체수평.결과 GD3、GD4、GD5화GD6각폭로조재잉후제7천관찰종점혈청E2수평유불동정도적하강,차이균유통계학의의(P,＜0.05)；GD3화GD5폭로조잉서재잉후제6천관찰종점기혈청E2수평야유소하강,차이유통계학의의(P＜0.05).Western blotting결과현시,GD4화GD5폭로조잉서자궁조직중ER-α균재폭로후제일개관찰종점(즉잉후제5천화잉후제6천)저표체,표체수평분별교대조조강저료23.6％화72.9％,차이균구유통계학의의(P＜0.05)；GD3화GD6폭로조잉서재각관찰종점ER-α표체수평균교대조조유소강저,단차이무통계학의의(P＞0.05).결론 배태식입기CS2폭로가강저자궁조직ER-α표체수평병감소잉서혈청E2함량,저가능시CS2치배태독성적분자궤제지일.
Objective To evaluate the effects of carbon disulfide (CS2) exposure during peri-implantation on the estrogen receptor-α (ER-α) expression in the uterus and serum level of estradiol (E2) in pregnant mice,and to explore the mechanism of embryotoxicity of CS2.Methods Healthy female mice were exposed to a single dose of CS2 (631.4 mg/kg) or olive oil (solvent control) on gestational day (GD)3,GD4,GDS,or GD6.At different time points after exposure,the serum E2 levels of the pregnant mice were measured by enzymelinked immunosorbent assay,and the expression levels of ER-α in the uterus were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot.Results Compared with the control group,the GD3,GD4,GD5,and GD6 exposure groups showed significantly decreased serum E2 levels on day 7 of gestation (P＜0.05); the GD3 and GD5 exposure groups showed significantly decreased serum E2 levels on day 6 of gestation (P＜0.05).The expression level of ER-α in the GD 4 exposure group was 23.6％ lower than that in the control group on day 5 of gestation,and the expression level of ER-o in the GD 5 exposure group was 72.9％ lower than that in the control group on day 6 of gestation (P＜0.05); the GD 3 and GD 6 exposure groups showed lower expression levels of ER-α than the control group at any time point,but no significant difference was found (P＞0.05).Conclusion CS2 exposure during peri-implantation can reduce the ER-α expression in the uterus and the serum level of E2 in pregnant mice,which may be one of the mechanisms of embryotoxicity of CS2.