5. 2
1. (DEP) 1) DEP 2) 3) 4) DEP 0 600 3000 15000 ppm(0 43 210 1083 mg/kg/day 0 54 261 1336 mg/kg/day ) 24 Crj:CD(SD) IGS 2 DEP 15000 ppm F0 0-21 F1 F0 F1 DEP F1 F1 15000 ppm 15000 ppm F0 F1 F0 F0 P-450 (CYP1A1/2 CYP2B1 CYP3A2 CYP4A1)15000 ppm CYP4A1 CYP3A2 CYP3A 6 5) F0 3000 15000 ppm DEP - 2 -
F1 F2 DEP 15000 ppm F1 4 21 F1 F2 21 F1 F2 DEP 15000 ppm F1 F2 4 F1 F2 DEP 3000 ppm F1 F2 15000 ppm F1 F2 F1 F2 F2 F1 F2 F1 F1 F2 F0 F1 15000 ppm 15000 ppm F0 P-450 CYP3A2 3000 15000 ppm F0 F0 F1 15000 ppm F1 F2 3000 ppm 15000 ppm 2 (NOEL) 3000 ppm(210 mg/kg/day 261 mg/kg/day )(NOAEL) 15000 ppm(1083mg/kg/day 1336 mg/kg/day ) 600 ppm(43 mg/kg/day 54 mg/kg/day ) 3000 ppm(210 mg/kg/day 261 mg/kg/day ) 3000 ppm - 3 -
600 ppm(43 mg/kg/day 54 mg/kg/day ) 15000 ppm(1083mg/kg/day 1336 mg/kg/day ) - 4 -
2 (DEP) 99.8 ( Lot No. SEM5441) 3 4 (NEDO) 2 5 Crj:CD(SD) IGS () 1 24 F0 5 6 F0 F1 15 17 (2000 10 26 2001 7 10 ) 7 0, 600, 3000 15000 ppm (NIH-07M) 8 2 ( ) 0 () 5000 10000 20000 40000 ppm 1 8 Crj:CD(SD)IGS 4 F0 20000 ppm 2 40000 ppm 1 40000 ppm 10000 ppm 20000 ppm 40000 ppm F1 20000 40000 ppm 40000 ppm 4 21 2 15000 ppm 5 3000 600 ppm 3 4-5 -
9 9.1 NIH-07M(H7010 H7020 9.2 9.3 23±3 50±20 10 15 12 8:00 20:00 9.4 300W 410D 200H mm, 1 1 17 21-6 -
10. 1 1 10.1 1 2 10.2 0 7 14 20 0 7 14 21 10.3 10 F0 15 F1 13 1 1 0 3 10.4 F1 F1 24 F1 5 21 25 1 F1 F1 24 2 F1 10.5 35 27 () ( ) () ( ) () 100-7 -
10.6 ( ) ( ) (HTM-IVOS Hamilton Thorne research ) ( 200 ) 10.7 P-450 6 F0 P-450 (CYP1A1/2 CYP2B1 CYP3A2 CYP4A1) 10.8 6 P-450 F0 DPC DPC RIA 10.9 0 4 7 14 21 0 4 0 ( ) ( 0 ) 100 4 ( ) ( 4 0 ) 100 21 ( ) ( 21 4 ) 100 10.10 1 4 8 12-8 -
10.11 1 4 7 13 10.12 F0 F1 ( ) 1 F1 F2 ( ) 10.13 4 F1 F1 F2 26 10.14 F0 F1 F1 6 F1 F2 6 F2 10.15 Bartlett - 9 -
Dunnett Kruskal-Wallis Mann-Whitney U 1 1 Wilcoxon 2 2 10 Fisher - 10 -
. F0 10 1 1 3 1:1 0 2 3 ( 0 7 14 20 ) ( 0-7 7-14 14-20 ) 0 3 4 8 ( 4 4 ) ( 0 7 14 21 ) ( 0-7 7-14 14-21 ) 0 4 7 14 21 0 4 ( 1 4 8 12 ) ( 4 7 13 ) - 11 -
F0/F1 F1 24 21 25 F1 ( 1 2 F1 F1 26 ) F0 6 P-450 1 ( ) 6 F1 10 1 35 27 1 3 1:1 0 F1 2 3 ( 0 7 14 20 ) ( 0-7 7-14 14-20 ) 0-12 -
F1 3 4 8 ( 4 4 ) ( 0 7 14 21 ) ( 0-7 7-14 14-21 ) 0 4 7 14 21 0 4 ( 1 4 8 12 ) ( 4 7 13 ) F1/F2 F2 26 ( ) 1 ( ) 6 6-13 -
- 14 -
11. 2. 11.1 11.1.1 2. Tables 1 and 2 F0F1 DEP DEP( )3000 ppmf0 1 ( ) F0 2 (1 1 ) DEP 15000 ppmf0f1 11.1.2 2. Tables 3 to 6 F0F1 15000 ppm3000ppm F0 600 ppm 15000 ppmf0 F10-1 F00-21 F121 F1 DEP 15000 ppmf0 F1 11.1.3 2. Tables 7 and 8 F0F13000 ppm F0 3000 ppm9100-7 7-14 15000 ppmf0f1 1 DEP DEP 15000 ppmf0f1-15 -
11.1.4 2. Tables 13, 14, and 16 F0F1 DEP F1 1)2. Table 13 F1 15000 ppm (22 23 ) 2) 2. Table 15 F0 3000 ppm F1 300015000 ppm Tailless DEP 3)2. Table 16 F1 15000 ppm 15000 ppmf1 DEP 15000 ppm F0F1 11.1.5 2. Tables 21 and 22 F0F115000 ppm3000 ppm 15000 ppmf0f1 F0 DEP ()F1 F1 DEP - 16 -
DEP 15000 ppm F0F1 F0 DEP 11.1.6 P-4502. Table 17 F0 P-450 (CYP1A1/2 CYP2B1 CYP3A2 CYP4A1) 3000 ppm15000 ppm CYP4A1CYP3A2 DEP P-450 CYP4A1P-450 6) 7) DEP CYP3A6 5) F0 DEP 15000 ppm P-450(CYP4A1 CYP3A2) P-450 11.1.7 2. Table 18 F0RIA 300015000 ppmcyp3a2 3000 ppm 15000 ppm 8) DEP 3000 ppm CYP3A2 11.1.8-17 -
2. Tables 19, 20, 23, and 24 F0F1 DEP15000 ppm DEP 15000 ppmf0 F1 11.2 11.2.1 2. Tables 25 and 26 F1F2DEP DEP 15000 ppmf1f2 11.2.2 2. Table 27 15000 ppmf1f2 F1 4 21 F1 F221 DEP 15000 ppmf1f2 11.2.3 2. Tables 13, 28, and 32 to 34 F1F204 21 DEP DEP 15000 ppmf1f2 11.2.4 2. Tables 29 to 31 F1F2 15000 ppm - 18 -
DEP 15000 ppm 11.2.5 2. Tables 35 and 36 F1 F24F1 F2 F1F1 F2 DEP DEP 15000 ppm F1F2 11.2.6 2. Tables 37 38 and 39 F1F2600 ppm F1 3000 ppmf1f2 15000 ppm 15000 ppmf1f2 F1F2 F1 F2 F1F2 F2 F1F2 F1 F1F2 DEP in vitro 2) in vivo 3) F1F1 F2 F1 F1 F2 DEP 3000 ppmf1f2 15000 ppm F1F2 F1F2 F2 F1F2 F1 F1F2-19 -
11.3 NOEL F0 F1 15000 ppm 15000 ppm F0 P-450 CYP3A2 3000 15000 ppm F0 F0 F1 15000 ppm F1 F2 3000 ppm 15000 ppm 2 (NOEL) 3000 ppm(210 mg/kg/day 261 mg/kg/day )(NOAEL) 15000 ppm(1083mg/kg/day 1336 mg/kg/day ) 600 ppm(43 mg/kg/day 54 mg/kg/day ) 3000 ppm(210 mg/kg/day 261 mg/kg/day ) 3000 ppm 600 ppm(43 mg/kg/day 54 mg/kg/day ) 15000 ppm(1083mg/kg/day 1336 mg/kg/day ) - 20 -
12. 1) 13398,,, 1998, pp 1017-1021. 2) Blair, R.M., Hong, F., William, S.B., Bruce, S.H., Stacey, L.D., Carrie, L.M., Weida, T., Leming, S., Roger, P., and Daniel, M.S. The estrogen receptor relative binding affinities of 188 natural and xenochemicals: structural diversity of ligands. Toxicol. Sci., 2000; 54: 138-153. 3) 2001 4) Harris, C.A., Henttu, P., Parker, M.G., Sumpter, J.P. The estrogenic activity of phthalate esters in vitro. Environ. Health Perspect., 1997; 105: 802-811. 5) Niwa, T., Yabusaki, Y., Honma, K., Matsuo, N., Tatsuta, K., Ishibashi, F. and Katagiri, M.: Contribution of human hepatic cytochrome P450 isoforms to regioselective hydroxylation of steroid hormones. Xenobiotica, 28, 539-547, 1998. 6) Bell, D.R. and Elcombe, C.R. Induction of acyl-coa oxidase and cytochrome P450 IV A1 RNA in rat primary hepatocyte culture by peroxisome proliferators. Biochemical Journal, 280, 249-253, 1991. 7) Okita, R.T. and Okita, J.R. Effects of diethyl phthalate and other plasticizers on laurate hydroxylation in rat liver microsomes. Pharmaceutical Research 9, 1648-1653, 1992. 8). Markert, C.L., 1979:87-95 - 21 -
2. F0() F1 F1 F2 ppm 0 600 3000 15000 0 600 3000 15000 24 24 24 24 24 24 24 24 24 24 24 24 24 24 24 24 mg/kg/day 40 197 1016 46 222 1150 51 255 1297 56 267 1375 0/24 0/24 1/24 0/24 0/24 0/24 0/24 0/24 2/24 0/24 0/24 0/24 0/24 0/24 0/24 0/24 (g) 427.862.5 416.855.9 428.332.3 403.951.8 536.248.4 535.559.4 548.874.7 552.565.5 f) [MeanS.D.] 150.722.6 161.122.1 a)b) 145.018.6 147.216.9 243.427.4 250.328.0 248.834.2 249.323.8 132.315.8 132.817.3 126.421.1 131.113.6 137.016.8 131.521.4 137.118.7 136.721.8 3.516.3-0.512.0 7.315.0 20.411.9 c) -19.719.9-17.313.3-19.425.1-12.222.4 (g) 23.55-29.42 24.01-30.01 23.67-29.15 23.38-29.26 13.62-31.17 13.74-31.56 14.57-30.99 14.88-32.50 g) 18.56-20.94 18.65-21.30 17.43-19.64 d) 18.09-20.27 13.25-22.46 12.96-23.39 13.47-22.17 13.98-22.23 g) 22.40-24.36 22.38-23.98 21.33-22.45 e) 22.02-23.26 23.93-25.62 24.28-25.81 23.83-24.65 23.76-25.36 38.30-73.16 35.85-74.12 36.29-66.49 38.46-66.56 35.76-65.77 36.19-69.33 34.58-65.66 36.14-64.16 42.51.8 43.02.7 42.41.8 42.61.7 [MeanS.D.] (g) 232.319.5 226.714.6 232.123.4 230.618.7 23/24 23/24 24/24 24/24 24/24 24/24 21/24 23/24 22/23 22/23 24/24 23/24 23/24 23/24 20/21 22/23 (x10 6 ) 230.6550.83 212.2670.20 215.8327.15 215.8533.50 226.8546.90 223.3541.87 225.2331.00 217.0036.11 (x10 6 ) 250.4050.17 230.9664.80 253.7840.52 242.0247.12 260.4041.83 246.2937.34 246.5432.36 254.9440.51 (%) 84.588.26 81.0921.02 85.1310.50 85.127.92 92.695.33 90.599.97 90.218.16 91.786.54 (%) 0.730.61 5.3519.69 1.260.96 0.690.76 0.600.53 0.830.72 1.310.84 1.521.18 30.01.5 31.12.2 30.72.0 31.72.1 (g) 105.013.3 108.411.5 108.39.1 110.312.2 23/24 23/24 22/24 24/24 23/24 24/24 20/24 21/24 ( )() 4.030.09 4.070.23 4.050.21 4.100.41 4.110.23 4.220.37 4.100.19 4.120.27 24/24 23/24 24/24 24/24 24/24 24/24 23/24 24/24 23/24 22/23 24/24 23/24 23/24 23/24 22/23 23/24 22/23 22/22 24/24 22/23 23/23 23/23 22/22 23/23 () 22.30.5 22.30.5 22.30.5 22.10.3 22.40.5 22.30.5 22.30.6 22.10.3 14.71.8 15.01.7 14.92.0 14.82.1 14.32.5 14.33.2 14.92.7 14.72.5 (%) 94.336.67 91.278.82 93.784.97 94.907.23 91.3311.07 93.688.42 91.8213.25 89.8017.89 13.92.0 13.72.0 14.02.1 14.02.2 13.02.8 13.53.5 13.93.1 13.33.5 a) 0-40-5 (p0.01)b) 0-80-9 (p0.05)c) 0-21 (p0.01)d) 910 (p0.05) e) 0-70-14 (p0.05)f) 0-1 (p0.01)g) 1 (p0.05) p0.05 p0.01
2. 1 F0() F1 F1 F2 ppm 0 600 3000 15000 0 600 3000 15000 24 24 24 24 24 24 24 24 24 24 24 24 24 24 24 24 a) (g) 18.9713.049 18.0362.434 18.8211.956 19.3682.512 19.2792.411 19.6032.880 20.1453.312 21.9364.053 [MeanS.D.] (mg) 27.043.09 26.544.86 28.235.74 26.606.46 25.355.15 26.965.01 29.835.40 26.855.12 (mg) 56.69.1 54.57.5 52.27.7 49.97.6 60.56.2 60.19.1 56.78.3 56.75.9 (g) 1.3020.130 1.2300.176 1.2920.077 1.2350.105 1.2760.149 1.2770.132 1.3190.105 1.2850.094 (%) 3.230.24 3.130.29 3.210.28 3.440.23 3.190.32 3.240.28 3.260.29 3.530.35 (x10-3 ) 4.640.56 4.620.73 4.800.90 4.721.03 4.190.79 4.490.92 4.880.98 4.370.89 (x10-3 ) 9.691.41 9.531.45 8.891.24 8.891.25 10.030.95 9.971.31 9.261.42 9.210.94 0.2250.027 0.2180.044 0.2210.017 0.2220.026 0.2120.026 0.2130.031 0.2160.028 0.2100.021 (g) 12.9562.086 12.9501.488 12.9951.442 14.4381.756 13.4281.332 14.1392.208 14.0331.745 14.8891.617 (g) 2.2310.269 2.2630.179 2.2330.189 2.3250.204 2.2620.218 2.2790.194 2.3240.241 2.4660.225 (%) 4.140.57 4.110.54 4.230.44 4.570.52 4.120.27 4.280.54 4.210.39 4.520.39 0.7130.068 0.7150.053 0.7280.054 0.7350.047 0.6960.056 0.6910.059 0.6980.060 0.7500.063 b) P-450 CYP3A2 55.620.5 62.218.5 33.213.1 92.015.4 (p mol/mg protein) CYP4A1 13.73.1 15.06.9 11.52.6 62.819.2 b) 2.5251.225 1.8100.460 0.5030.258 1.2620.656 (ng/ml) 2.6622.514 3.6052.496 5.9802.996 3.0820.941 p0.05 p0.01
2. 2 F0() F1 F1 F2 ppm 0 600 3000 15000 0 600 3000 15000 22 22 24 22 23 23 22 23 21 22 23 21 23 23 21 22 0.544 0.465 0.507 0.490 0.510 0.510 0.523 0.482 (%) 0 97.57 98.82 97.78 97.20 96.24 98.60 97.92 98.57 4 97.77 96.24 97.24 97.71 96.34 97.80 99.37 98.09 21 97.73 95.45 90.63 88.07 94.02 96.74 98.86 99.46 0-21 0-21 (g) 0 6.850.74 6.970.64 6.990.75 6.700.66 7.060.52 6.990.72 6.850.69 6.760.56 [MeanS.D.] 4 10.621.39 10.101.40 10.501.86 9.661.36 11.391.83 10.601.51 10.681.22 10.801.57 7 17.362.37 15.632.61 16.003.77 15.303.24 18.263.01 17.472.66 18.071.53 17.832.21 14 35.563.75 33.623.31 33.376.00 32.174.76 36.476.39 36.133.86 36.823.06 36.393.48 21 59.037.70 56.816.59 55.139.53 48.104.81 60.869.50 61.406.87 60.564.53 53.754.85 0 6.550.69 6.540.68 6.540.72 6.260.59 6.630.64 6.530.68 6.440.47 6.290.40 4 10.211.38 9.551.51 9.931.62 9.101.13 10.762.07 9.941.39 9.990.94 9.981.09 7 16.672.40 15.402.76 15.433.18 14.392.74 17.053.69 16.802.37 17.141.70 16.741.59 14 34.333.77 33.274.07 32.784.50 30.006.06 35.106.42 35.073.92 35.712.92 34.802.72 21 56.276.58 55.387.05 53.777.21 45.854.46 58.179.41 58.726.23 58.164.23 51.514.16 0 3.6700.306 3.7220.293 3.8340.351 3.5640.280 3.6340.292 3.5950.168 3.6400.201 3.6500.237 (mm) 4 5.6580.496 5.4570.442 5.5440.690 5.5440.554 5.7820.420 5.6570.385 5.7290.356 5.7730.491 [MeanS.D.] 0 1.7580.207 1.6670.218 1.6720.234 1.7740.216 1.7630.111 1.7310.089 1.7650.091 1.7360.089 4 2.7690.337 2.7030.250 2.7210.344 2.7380.327 2.9270.232 2.8940.218 2.9020.215 2.8810.202 2 /3 40.5/98.1 45.1/92.2 47.7/92.9 14.9/82.4 53.3/88.3 47.4/85.9 33.6/95.5 23.1/94.2 (, %) 10 /11 4.9/37.2 13.6/45.1 17.5/49.2 23.5/65.5 18.6/50.1 25.3/60.7 11.9/35.0 15.6/44.6 12 /13 70.3/93.6 72.0/90.9 79.4/96.5 85.2/96.6 92.2/98.9 75.5/91.9 77.2/92.0 79.0/96.4 13 /14 8.2/70.3 9.1/62.8 4.2/56.5 11.5/47.6 18.5/63.6 24.0/68.0 15.9/62.4 10.9/65.2 2 /3 39.1/97.3 46.3/86.2 44.4/93.3 17.8/82.8 55.0/86.2 42.7/91.6 35.3/95.9 20.5/93.6 10 /11 12.9/43.0 23.9/62.8 29.2/61.0 30.7/65.2 25.0/58.1 23.9/51.8 14.7/47.4 16.8/52.5 12 /13 77.2/100 78.7/95.9 90.6/96.9 80.3/93.2 92.4/100 72.5/90.2 84.4/96.7 82.0/96.6 13 /14 16.8/76.9 22.0/75.0 14.2/70.1 13.6/48.9 28.8/66.3 21.7/76.1 15.5/68.3 9.1/80.0 15 100 95.5 99.0 88.6 91.7 97.8 98.1 98.0 4.00.0 4.00.0 4.00.0 4.00.0 4.00.0 4.00.0 4.00.0 4.00.0 ( ) 7.70.9 7.30.6 7.40.8 7.51.0 7.20.4 7.50.9 7.40.5 7.50.7 [MeanS.D.] 16.21.3 16.51.3 16.21.7 16.21.8 16.31.2 16.11.4 16.11.4 15.61.4 4.00.2 4.10.3 4.10.3 4.20.4 4.00.2 4.00.0 4.00.0 4.00.0 7.30.6 7.61.1 7.30.6 7.50.9 7.40.6 7.10.3 7.50.7 7.10.4 16.21.5 16.21.3 16.32.0 16.32.2 16.40.9 16.51.0 16.01.4 16.21.6 p0.05 p0.01
2. 3 F0() F1 F1 F2 ppm 0 600 3000 15000 0 600 3000 15000 22 22 24 22 23 23 22 23 21 22 23 21 23 23 21 22 a) (g) 3.9050.473 3.7050.422 3.8290.683 3.7530.513 3.8900.692 3.8940.538 4.0050.439 4.2210.527 [MeanS.D.] (g) 0.9940.101 1.0160.121 0.9890.155 0.8940.084 0.9770.136 0.9990.124 0.9940.090 0.9180.101 (g) 0.3580.053 0.3420.053 0.3570.073 0.3180.037 0.3490.061 0.3250.060 0.3530.063 0.2980.058 (g) 1.6210.048 1.6350.067 1.6310.092 1.5940.082 1.6550.100 1.6630.062 1.6670.050 1.6410.063 (mg) 3.390.44 3.400.48 3.480.59 3.300.58 3.410.74 3.501.00 3.530.61 3.270.60 (mg) 352.247.6 340.371.5 354.587.8 270.452.4 367.164.0 375.260.9 357.654.1 292.053.8 (mg) 25.63.1 26.33.0 24.03.7 22.54.3 25.35.2 24.62.8 24.63.1 22.03.8 (mg) 40.010.2 40.09.3 39.99.0 32.39.2 42.912.5 43.510.0 42.810.5 37.88.8 (mg) 19.63.0 19.73.5 20.24.5 19.73.6 20.04.9 18.73.9 19.13.5 18.03.1 4.630.24 4.400.25 4.570.22 5.150.21 4.420.27 4.410.24 4.560.32 5.140.29 1.180.07 1.210.11 1.190.06 1.230.09 1.120.07 1.130.06 1.130.07 1.120.06 0.4250.046 0.4080.050 0.4270.056 0.4390.041 0.4050.070 0.3670.047 0.4010.053 0.3620.046 1.940.17 1.960.15 1.990.26 2.210.22 1.940.36 1.910.18 1.910.15 2.020.20 (x10-3 ) 4.050.59 4.050.44 4.190.49 4.560.73 3.940.82 3.960.93 4.030.66 4.010.76 (x10-3 ) 42053 40578 42262 37258 42043 42652 40645 35654 (x10-3 ) 30.53.7 31.42.9 29.13.6 31.04.7 28.83.5 28.13.4 28.13.5 26.94.3 (x10-3 ) 47.410.4 47.59.4 47.98.6 44.210.6 49.011.8 49.511.0 48.811.5 45.99.1 (x10-3 ) 23.43.8 23.74.8 24.45.3 27.35.2 23.04.6 21.34.3 21.94.3 22.13.8 (g) 3.4100.539 3.1480.427 3.1980.629 3.0040.388 3.4100.490 3.4600.429 3.5250.426 3.6750.273 (g) 0.9450.097 0.9240.111 0.9300.139 0.7870.089 0.9150.101 0.9240.100 0.9080.071 0.8420.059 (g) 1.5980.071 1.5890.084 1.5970.085 1.5200.105 1.6100.079 1.6130.064 1.6050.067 1.5970.064 (mg) 362.450.0 353.453.6 361.759.1 254.644.9 370.163.1 372.058.9 347.861.6 299.548.7 (mg) 7.751.49 7.431.12 7.401.01 6.671.03 7.891.53 8.091.75 8.161.66 7.321.23 (mg) 25.84.6 25.84.2 22.83.4 20.63.5 23.64.3 23.03.3 23.33.4 20.33.3 (mg) 58.811.5 60.915.4 63.116.1 45.810.9 63.322.0 55.714.4 52.49.9 46.310.8 (%) 4.220.26 4.030.34 4.090.37 4.600.23 4.150.29 4.170.32 4.230.34 4.810.26 1.180.05 1.180.09 1.200.08 1.210.09 1.120.07 1.120.08 1.090.05 1.100.06 2.000.17 2.050.18 2.080.24 2.350.22 1.990.26 1.960.18 1.940.14 2.100.12 (x10-3 ) 45148 45570 46761 38947 45147 44850 41861 39154 (x10-3 ) 9.722.07 9.591.74 9.631.54 10.311.76 9.701.89 9.802.08 9.842.02 9.601.57 (x10-3 ) 32.04.8 33.14.9 29.64.3 31.53.7 28.84.3 27.93.4 28.13.6 26.74.3 (x10-3 ) 74.218.0 78.318.8 81.017.8 69.913.8 76.322.5 66.713.2 63.010.1 60.613.8 p0.05 p0.01