CAJ | 학술논문

目的研究胚胎植入期二硫化碳(CS2)暴露对孕鼠脾脏淋巴细胞DNA损伤的影响,从免疫损伤角度探讨CS2致胚胎植入障碍的机制.方法分别建立不同暴露剂量和不同暴露时间2个动物模型:模型1为不同CS2暴露剂量,即在小鼠受孕第4天(GD4),分别给予4组小鼠一次性腹腔注射低(0.1LD50,157.8 mg/kg)、中(0.2LD50,315.7 mg/kg)、高(0.4LD50,631.4 mg/kg)剂量的CS2和橄榄油对照组；模型2为不同CS2暴露时间,即4组小鼠分别在GD3、GD4、GD5和GD6一次性腹腔注射CS2(0.4LD50,631.4 mg/kg),各组设平行对照.两模型对照组均注射等体积橄榄油.实验终点,制备胚胎植入期小鼠脾脏淋巴细胞单细胞悬液,用台盼蓝法检测细胞活性,并进行碱性单细胞凝胶电泳(SCGE)试验,检测孕鼠脾脏淋巴细胞DNA损伤状况.结果 (1)中、高剂量组的DNA损伤程度与对照组相比差异均有统计学意义(P＜0.01)；反映DNA损伤程度的各指标与暴露剂量之间均存在回归关系,且差异有统计学意义(P＜0.01).(2)GD3、GD4、GD5和GD6暴露组的DNA损伤程度与对照组比较,差异均有统计学意义(P＜0.01),且GD4暴露组DNA损伤程度最明显.结论胚胎植入期CS2暴露可导致孕鼠脾脏淋巴细胞的DNA损伤,并且呈明显的剂量反应关系；GD4可能是CS2暴露致孕鼠脾脏淋巴细胞DNA损伤的敏感时间点.
목적 연구배태식입기이류화탄(CS2)폭로대잉서비장림파세포DNA손상적영향,종면역손상각도탐토CS2치배태식입장애적궤제.방법 분별건립불동폭로제량화불동폭로시간2개동물모형:모형1위불동CS2폭로제량,즉재소서수잉제4천(GD4),분별급여4조소서일차성복강주사저(0.1LD50,157.8 mg/kg)、중(0.2LD50,315.7 mg/kg)、고(0.4LD50,631.4 mg/kg)제량적CS2화감람유대조조；모형2위불동CS2폭로시간,즉4조소서분별재GD3、GD4、GD5화GD6일차성복강주사CS2(0.4LD50,631.4 mg/kg),각조설평행대조.량모형대조조균주사등체적감람유.실험종점,제비배태식입기소서비장림파세포단세포현액,용태반람법검측세포활성,병진행감성단세포응효전영(SCGE)시험,검측잉서비장림파세포DNA손상상황.결과 (1)중、고제량조적DNA손상정도여대조조상비차이균유통계학의의(P＜0.01)；반영DNA손상정도적각지표여폭로제량지간균존재회귀관계,차차이유통계학의의(P＜0.01).(2)GD3、GD4、GD5화GD6폭로조적DNA손상정도여대조조비교,차이균유통계학의의(P＜0.01),차GD4폭로조DNA손상정도최명현.결론 배태식입기CS2폭로가도치잉서비장림파세포적DNA손상,병차정명현적제량반응관계；GD4가능시CS2폭로치잉서비장림파세포DNA손상적민감시간점.
Objective To investigate the DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide (CS2) in the implantation phase and to explore the mechanism of abnormal implantation induced by CS2 from the perspective of immune injury.Methods Mice were exposed to CS2 at different doses or at different time points in the implantation phase to establish model 1 and model 2.For model 1,mice were assigned to four groups to receive a single intraperitoneal injection of low-dose CS2 (0.1 LD50,157.8mg/kg),middle-dose CS2 (0.2 LD50,315.7 mg/kg),and high-dose CS2 (0.4 LD50,631.4 mg/kg) as well as an equal volume of olive oil (control) on gestational day (GD) 4.For model 2,mice were assigned to four groups to receive a single intraperitoneal injection of CS2 (0.4 LD50,631.4 mg/kg) or an equal volume of olive oil (control) on GD3,GD4,GD5,and GD6.At the end,single cell suspension of splenic lymphocytes was prepared.Cell viability was measured by trypan blue staining,and the DNA damage of splenic lymphocytes was evaluated by alkaline single cell gel electrophoresis assay.Results The middle-dose and high-dose exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P＜0.01); there was significant regression relationship between indicators of DNA damage and exposure doses (P＜0.01).The GD3,GD4,GDS,and GD 6 exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P＜0.01),and the GD 4 exposure group had the most DNA damage.Conclusion Exposure to CS2 in the implantation phase can induce DNA damage of splenic lymphocytes in pregnant mice,and the DNA damage was aggravated with the increase in CS2 concentration.GD4 may be the sensitive time point for DNA damage of splenic lymphocytes induced by CS2 in pregnant mice.