0 10 Inhibitory inhibitory JJ 11 17 effect effect NN 18 20 of of IN 21 26 E3330 E3330 NNP 26 27 , , , 28 29 a a DT 30 35 novel novel JJ 36 43 quinone quinone NN 44 54 derivative derivative NN 55 59 able able JJ 60 62 to to TO 63 71 suppress suppress VB 72 77 tumor tumor NN 78 86 necrosis necrosis NN 87 99 factor-alpha factor-alpha NN 100 110 generation generation NN 110 111 , , , 112 114 on on IN 115 125 activation activation NN 126 128 of of IN 129 136 nuclear nuclear JJ 137 149 factor-kappa factor-kappa NN 150 152 B. B. NNP 154 155 ( ( ( 155 217 2E)-3-[5-(2,3-Dimethoxy-6-methyl-1,4-benzoquinoyl)]-2-nonyl-2- 2e)-3-[5-(2,3-dimethoxy-6-methyl-1,4-benzoquinoyl)]-2-nonyl-2- JJ 218 227 propenoic propenoic JJ 228 232 acid acid NN 233 234 ( ( ( 234 239 E3330 E3330 NNP 239 240 ) ) ) 240 241 , , , 242 244 is be VBZ 245 246 a a DT 247 252 novel novel JJ 253 258 agent agent NN 259 263 with with IN 264 280 hepatoprotective hepatoprotective JJ 281 289 activity activity NN 289 290 . . . 291 293 We we PRP 294 300 report report VBP 301 304 the the DT 305 311 effect effect NN 312 314 of of IN 315 320 E3330 e3330 NN 321 323 on on IN 324 339 transcriptional transcriptional JJ 340 350 activation activation NN 351 353 of of IN 354 359 tumor tumor NN 360 368 necrosis necrosis NN 369 375 factor factor NN 376 377 ( ( ( 377 387 TNF)-alpha TNF)-alpha NNP 388 392 gene gene NN 393 396 and and CC 397 399 on on IN 400 407 nuclear nuclear JJ 408 414 factor factor NN 415 416 ( ( ( 416 425 NF)-kappa NF)-kappa NNP 426 427 B B NNP 428 438 activation activation NN 438 439 . . . 440 447 Nuclear nuclear JJ 448 454 run-on run-on JJ 455 466 experiments experiment NNS 467 473 showed show VBD 474 478 that that IN 479 484 E3330 e3330 NN 485 494 decreases decrease VBZ 495 510 transcriptional transcriptional JJ 511 521 activation activation NN 522 524 of of IN 525 534 TNF-alpha TNF-alpha NNP 535 539 gene gene NN 540 547 induced induce VBN 548 550 by by IN 551 569 lipopolysaccharide lipopolysaccharide NN 570 571 ( ( ( 571 574 LPS LPS NNP 574 575 ) ) ) 576 587 stimulation stimulation NN 588 590 in in IN 591 596 human human JJ 597 607 peripheral peripheral JJ 608 617 monocytes monocyte NNS 617 618 . . . 619 621 To to TO 622 633 investigate investigate VB 634 637 the the DT 638 648 inhibitory inhibitory JJ 649 659 mechanisms mechanism NNS 659 660 , , , 661 663 we we PRP 664 675 constructed construct VBD 676 677 a a DT 678 691 secreted-type secreted-type JJ 692 701 placental placental JJ 702 710 alkaline alkaline NN 711 722 phosphatase phosphatase NN 723 724 ( ( ( 724 728 PLAP PLAP NNP 728 729 ) ) ) 730 738 reporter reporter NN 739 743 gene gene NN 744 749 whose whose WP$ 750 763 transcription transcription NN 764 766 is be VBZ 767 777 controlled control VBN 778 780 by by IN 781 782 a a DT 783 789 1.4-kb 1.4-kb JJ 790 795 human human JJ 796 805 TNF-alpha TNF-alpha NNP 806 814 promoter promoter NN 814 815 . . . 816 817 A a DT 818 824 stable stable JJ 825 837 transformant transformant NN 838 840 of of IN 841 844 the the DT 845 849 PLAP PLAP NNP 850 858 reporter reporter NN 859 863 gene gene NN 864 871 derived derive VBN 872 876 from from IN 877 882 human human JJ 883 892 monocytic monocytic JJ 893 897 cell cell NN 898 902 line line NN 903 909 showed show VBD 910 914 very very RB 915 921 little little JJ 922 930 activity activity NN 931 933 on on IN 934 937 the the DT 938 946 promoter promoter NN 947 953 before before IN 954 965 stimulation stimulation NN 965 966 , , , 967 974 whereas whereas IN 975 978 LPS LPS NNP 979 990 stimulation stimulation NN 991 994 led lead VBD 995 997 to to TO 998 999 a a DT 1000 1008 dramatic dramatic JJ 1009 1017 increase increase NN 1018 1020 in in IN 1021 1025 PLAP PLAP NNP 1026 1034 activity activity NN 1034 1035 . . . 1036 1041 E3330 E3330 NNP 1042 1051 inhibited inhibit VBD 1052 1056 this this DT 1057 1064 induced induce VBN 1065 1073 promoter promoter NN 1074 1082 activity activity NN 1083 1085 in in IN 1086 1087 a a DT 1088 1102 dose-dependent dose-dependent JJ 1103 1109 manner manner NN 1109 1110 . . . 1111 1116 There there EX 1117 1120 are be VBP 1121 1125 four four CD 1126 1134 putative putative JJ 1135 1143 NF-kappa NF-kappa NNP 1144 1145 B B NNP 1146 1153 binding binding NN 1154 1159 sites site NNS 1160 1161 ( ( ( 1161 1166 kappa kappa NN 1167 1170 B-1 b-1 NN 1170 1171 , , , 1172 1177 kappa kappa NN 1178 1181 B-2 b-2 NN 1181 1182 , , , 1183 1188 kappa kappa NN 1189 1192 B-3 B-3 NNP 1192 1193 , , , 1194 1199 kappa kappa NN 1200 1203 B-4 b-4 NN 1203 1204 ) ) ) 1205 1207 in in IN 1208 1213 human human JJ 1214 1223 TNF-alpha TNF-alpha NNP 1224 1232 promoter promoter NN 1232 1233 . . . 1234 1236 By by IN 1237 1242 using use VBG 1243 1250 mutated mutate VBN 1251 1264 promoter-PLAP promoter-plap NN 1265 1273 plasmids plasmid NNS 1273 1274 , , , 1275 1277 we we PRP 1278 1289 established establish VBD 1290 1294 that that IN 1295 1300 these these DT 1301 1309 NF-kappa NF-kappa NNP 1310 1311 B B NNP 1312 1317 sites site NNS 1318 1322 were be VBD 1323 1332 necessary necessary JJ 1333 1336 for for IN 1337 1346 induction induction NN 1347 1349 of of IN 1350 1359 TNF-alpha TNF-alpha NNP 1360 1373 transcription transcription NN 1374 1376 on on IN 1377 1388 stimulation stimulation NN 1389 1393 with with IN 1394 1397 LPS LPS NNP 1397 1398 . . . 1399 1400 A a DT 1401 1404 gel gel NN 1405 1416 retardation retardation NN 1417 1427 experiment experiment NN 1428 1432 with with IN 1433 1442 synthetic synthetic JJ 1443 1458 double-stranded double-stranded JJ 1459 1475 oligonucleotides oligonucleotide NNS 1476 1482 showed show VBD 1483 1487 that that IN 1488 1497 activated activate VBN 1498 1506 NF-kappa NF-kappa NNP 1507 1508 B B NNP 1509 1519 consisting consist VBG 1520 1522 of of IN 1523 1530 p50/p65 p50/p65 NN 1531 1542 heterodimer heterodimer NN 1543 1548 bound bind VBD 1549 1551 to to TO 1552 1555 all all DT 1556 1560 four four CD 1561 1569 putative putative JJ 1570 1578 NF-kappa NF-kappa NNP 1579 1580 B B NNP 1581 1584 DNA DNA NNP 1585 1591 probes probe NNS 1591 1592 , , , 1593 1603 suggesting suggest VBG 1604 1608 that that IN 1609 1612 all all DT 1613 1617 four four CD 1618 1626 putative putative JJ 1627 1635 NF-kappa NF-kappa NNP 1636 1637 B B NNP 1638 1649 recognition recognition NN 1650 1655 sites site NNS 1656 1660 play play VBP 1661 1663 an an DT 1664 1673 important important JJ 1674 1678 role role NN 1679 1681 in in IN 1682 1691 inducible inducible JJ 1692 1701 TNF-alpha TNF-alpha NNP 1702 1712 expression expression NN 1712 1713 . . . 1714 1719 E3330 E3330 NNP 1720 1729 decreased decrease VBD 1730 1739 activated activate VBN 1740 1748 NF-kappa NF-kappa NNP 1749 1750 B B NNP 1751 1753 in in IN 1754 1760 nuclei nucleus NNS 1760 1761 , , , 1762 1772 suggesting suggest VBG 1773 1777 that that IN 1778 1783 E3330 e3330 NN 1784 1792 inhibits inhibit VBZ 1793 1801 NF-kappa NF-kappa NNP 1802 1803 B B NNP 1804 1814 activation activation NN 1815 1821 and/or and/or CC 1822 1835 translocation translocation NN 1836 1838 of of IN 1839 1842 the the DT 1843 1849 nuclei nucleus NNS 1849 1850 . . . 1851 1858 Western western NN 1859 1867 blotting blotting NN 1868 1876 analysis analysis NN 1877 1881 with with IN 1882 1888 anti-I anti-i JJ 1889 1894 kappa kappa NN 1895 1902 B-alpha B-alpha NNP 1903 1911 antibody antibody NN 1912 1921 indicated indicate VBD 1922 1926 that that IN 1927 1932 E3330 E3330 NNP 1933 1942 inhibited inhibit VBD 1943 1954 degradation degradation NN 1955 1957 of of IN 1958 1959 I i NN 1960 1965 kappa kappa NN 1966 1973 B-alpha B-alpha NNP 1973 1974 , , , 1975 1980 which which WDT 1981 1983 is be VBZ 1984 1986 an an DT 1987 1997 inhibitory inhibitory JJ 1998 2005 protein protein NN 2006 2008 of of IN 2009 2017 NF-kappa NF-kappa NNP 2018 2019 B B NNP 2019 2020 , , , 2021 2023 in in IN 2024 2038 LPS-stimulated lps-stimulated JJ 2039 2048 monocytes monocyte NNS 2048 2049 . . . 2050 2055 E3330 E3330 NNP 2056 2059 may may MD 2060 2068 suppress suppress VB 2069 2072 the the DT 2073 2083 production production NN 2084 2086 of of IN 2087 2093 active active JJ 2094 2100 oxygen oxygen NN 2101 2108 species specie NNS 2109 2116 serving serve VBG 2117 2119 as as IN 2120 2126 common common JJ 2127 2137 messengers messenger NNS 2138 2140 to to TO 2141 2149 activate activate VB 2150 2158 NF-kappa NF-kappa NNP 2159 2160 B B NNP 2160 2161 . . .