农药敌匹硫磷和残杀威与内分泌干扰物双酚A联合暴露对细胞吞噬功能的影响

Influence of joint exposure to diazinon, propoxur and bisphenol A on phagocytosis of RAW264. 7 cell

目的 探讨农药敌匹硫磷和残杀威与环境内分泌干扰物双酚A对细胞吞噬功能毒效应的联合作用方式.方法 采用荧光微球流式细胞仪检测技术,以吞噬百分率（PP）和吞噬指数（PI）为检测指标,观察无细胞毒性剂量的敌匹硫磷和双酚A、残杀威和双酚A联合暴露（受试物分别按等毒性比混合）对小鼠巨噬细胞RAW264.7吞噬功能的影响.敌匹硫磷与双酚A联合暴露终浓度分别为（0.4＋0.1）、（3.6＋0.7）、（36.2＋7.2）、（43.4＋8.7）、（52.1＋10.4）、（62.5＋12.5）、（75.0＋15.0）μg/ml;残杀威与双酚A联合暴露终浓度分别为（0.2＋2.0×10-2）、（2.4＋0.2）、（23.7＋2.0）、（35.6＋3.0）、（53.3＋4.4）、（80.0＋6.7）、（120.0＋10.0）μg/ml.以剂量-效应关系为基础,选择效应差异具有统计学意义的敌匹硫磷与双酚A、残杀威与双酚A剂量分别进行2×2完全析因实验,分析敌匹硫磷与双酚A、残杀威与双酚A的联合作用方式.结果 在联合暴露条件下,与对照组[PP为（23.6±2.2）%;PI为0.36±0.03]相比,敌匹硫磷＋双酚A各剂量组[（52.1＋10.4）、（62.5＋12.5）、（75.0＋15.0）μg/ml]均能提高细胞的PP[（29.0±1.4）%,t=3.89,P〈0.05;（30.2±2.3）%,t=4.74,P〈0.05;（35.0±3.4）%,t=8.21,P〈0.05]和PI（0.43±0.03,t=3.86,P〈0.05;0.41±0.02,t=2.95,P〈0.05;0.46±0.03,t=5.34,P〈0.05）水平;而残杀威＋双酚A各剂量组[（35.6＋3.0）、（53.3＋4.4）、（80.0＋6.7）、（120.0＋10.0）μg/ml]则降低细胞的PP[（20.6±1.1）%,t=-3.00,P〈0.05;（20.2±1.0）%,t=-3.42,P〈0.05;（19.4±1.3）%,t=-4.23,P〈0.05;（18.8±2.1）%,t=-4.81,P〈0.05]和PI（0.31±0.01,t=-4.75,P〈0.05;0.31±0.01,t=-4.58,P〈0.05;0.30±0.01,t=-4.92,P〈0.05;0.27±0.02,t=-7.80,P〈0.05）水平.选择敌匹硫磷[60.0μg/ml,PP为（28.5±3.4）%,PI为0.49±0.07]与双酚A[12.0μg/ml,PP为（35.7±2.7）%,PI为0.67±0.07]、残杀威[48.0 μg/ml,PP为（28.1±2.2）%,PI为0.48±0.04]与双酚A[4.0μg/m1,PP为（34.4±2.7）%,PI为0.59±0.07],分别进行2×2析因设计实验.分析结果显.示,敌匹硫磷和双酚A对RAW264.7细胞的PP[（30.4±1.4）%]和PI（0.53±0.03）的影响存在交互作用（PP,F交互作用=6.22,P〈0.05;PI,F交互作用=7.35,P〈0.05）,残杀威和双酚A之间亦存在交互作用[PP为（27.5±4.1）%,F交互作用=4.56,P〈0.05;PI为0.46±0.08,F交互作用=11.13,P〈0.05],均表现为拮抗作用.结论 敌匹硫磷与双酚A、残杀威与双酚A对RAW264.7细胞吞噬功能毒效应的联合作用方式均为拮抗作用.

Objective To explore the toxicity of joint exposure to diazinon,propoxur and bisphenol A on phagocytosis. Methods Flow cytometer was employed to detect the influence of diazinon and bisphenol A, propoxur and bisphenol A in mixture （mixed according to ratio of IC50） on mouse macrophage RAW264. 7 cells＇ function to phagocyte fluorescent microspheres, adopting the percentage of phagocytic cells （PP） and the phagocytic index （PI） as measurement indicators. The final concentrations of mixture of diazinou and bisphenol A were （0. 4 ＋ 0. 1）, （3.6 ＋ 0. 7）, （36. 2 ＋ 7. 2）, （43.4 ＋ 8.7）, （52.1 ＋ 10.4）,（62.5 ＋ 12.5）, （75.0 ＋ 15.0） μg/ml; while those of mixture of propoxur and bisphenol A were （0.2＋2.0×10-2）,（2.4 ＋0.2）, （23.7 ＋2.0）,（35.6 ＋3.0）,（53.3 ＋4.4）,（80.0 ＋6.7）,（120. 0 ＋ 10.0） μg/ml. Then based on the dose-response relationship, a 2 × 2 factorial design was then carried out among different doses of mixture with statistical significance to statistically evaluate the interaction between diazinon and bisphenol A, propoxur and bisphenol A. Results After the joint exposure, compared to the control group （PP = （23.6 ± 2. 2） %; PI = 0. 36 ± 0. 03）, any dose of the mixture of diazinon and bisphenol A （（52. 1 ＋ 10. 4）, （62. 5 ＋ 12. 5）, （75.0 ＋ 15.0） μg/ml） could significantly increase the levels of PP （（29.0±1.4）%,t=3.89,P〈0. 05; （30.2 ±2.3）%,t =4.74,P〈0.05; （35.0±3.4）%,t=8.21,P 〈0. 05） and PI （0.43 ±0. 03,t=3.86,P 〈0. 05; 0.41 ±0. 02,t=2.95,P 〈0. 05; 0.46 ±0. 03, t = 5. 34, P 〈 0. 05）; while that of propoxur and bisphenol A （（35.6 ＋ 3.0）, （53.3 ＋ 4. 4）, （80. 0 ＋6.7） ,（120.0 ＋ 10.0） μg/ml） reduced the levels of PP （（20.6±1.1）% ,t=-3.00,P〈0.05; （20.2±1.0）%,t=-3.42,P〈0.05; （19.4±1.3）%,t=-4.23,P〈0.05; （18.8±2.1）%,t=-4.81,P〈0.05） and PI （0.31 ±0.01,t =-4.75,P〈0.05; 0.31 ±0.01,t =-4.58,P〈0.05; 0.30 ±0.01,t=-4. 92, P 〈 0. 05; 0. 27 ± 0. 02, t =-7. 80, P 〈 0. 05） on the contrary. The 2 × 2 factorial design was carried out between the mixture of diazinon （60. 0 μg/ml; PP = （28. 5 ± 3.4） %; PI = 0. 49 ± 0. 07） and bisphenol A （12.0 μg/ml; PP= （35.7 ± 2.7）%; PI= 0.67 ± 0.07）, and the mixture of propoxur （48.0 μg/ml; PP= （28. 1 ±2.2）%; PI=0.48 ±0.04） and bisphenol A （4.0 μg/ml; PP= （34.4 ±2. 7） %; PI = 0. 59 ± 0. 07）. The mixture of diazinon and bisphenol A （PP = （30. 4 ± 1.4） %, Finteraction=6. 22, P 〈 0.05; PI = 0. 53 ± 0. 03, Finteraction = 7. 35, P 〈 0. 05） and the mixture of propoxur and bisphenol A （PP= （27.5 ±4.1）%,Finteraction=4.56,P〈0.05; PI = 0. 46 ± 0. 08, Finteraction=11.13,P〈0.05） both showed a significant antagonistic interaction on phagocytosis of RAW264. 7 cell. Conclusion It is suggested that the interactions between diazinon ＆ bisphenol A and propoxur ＆ bisphenol A both played the antagonistic role on phagocytic function of macrophages in vitro.