#METABOLOMICS WORKBENCH DongHo_20160115_9800591_mwtab.txt DATATRACK_ID:485 DongHo_20160115_025706 STUDY_ID:ST000316 ANALYSIS_ID:AN000502 PROJECT_ID:PR000255 VERSION 1 CREATED_ON January 15, 2016, 2:57 am #PROJECT PR:PROJECT_TITLE Comparison of Metabolites Variation and Antiobesity Effects of a Mixture of PR:PROJECT_TITLE Cudrania tricuspidata, Lonicera caerulea, and the Soybean According to PR:PROJECT_TITLE Fermentation in vitro and in vivo PR:PROJECT_SUMMARY We used ultra-performance-liquid-chromatography with quadrupole-time-of-flight PR:PROJECT_SUMMARY mass spectrometry to study the changes in metabolites in the mixture of Cudrania PR:PROJECT_SUMMARY tricuspidata, Lonicera caerulea, and soybean (CLM) during fermentation. PR:PROJECT_SUMMARY Additionally, the antiobesity effects of CLM and fermented-CLM (FCLM) were PR:PROJECT_SUMMARY studied based on the analysis of plasma from high-fat diet (HFD)-fed mice. The PR:PROJECT_SUMMARY levels of cyanidin and the glycosides of luteolin, quercetin, and cyanidin PR:PROJECT_SUMMARY derived from L. caerulea were decreased, whereas the levels of luteolin and PR:PROJECT_SUMMARY quercetin were increased during fermentation. Isoflavone glycosides and PR:PROJECT_SUMMARY soyasaponins originating from the soybean were decreased, whereas their PR:PROJECT_SUMMARY aglycones such as daidzein, glycitein, and genistein were increased. As for PR:PROJECT_SUMMARY prenylated flavonoids from C. tricuspidata, these metabolites were decreased at PR:PROJECT_SUMMARY the early stage of fermentation, and were increased at end of the fermentation. PR:PROJECT_SUMMARY In terms of the functional food product, various metabolites derived from PR:PROJECT_SUMMARY diverse natural products in CLM had complementary effects and demonstrated PR:PROJECT_SUMMARY higher antioxidant and pancreatic lipase inhibition activities by fermentation; PR:PROJECT_SUMMARY these activities were closely related to flavonoid aglycones including PR:PROJECT_SUMMARY genistein, daidzein, glycitein, luteolin, and quercetin. In vivo experiment, PR:PROJECT_SUMMARY several clinical parameters affected by HFD were remarkably improved by the PR:PROJECT_SUMMARY administration of either CLM or FCLM, but there was a difference in the PR:PROJECT_SUMMARY antiobesity effects. The levels of lysoPCs with C20:4, C16:0, and C22:6 were PR:PROJECT_SUMMARY significantly attenuated by CLM administration, while the attenuated levels of PR:PROJECT_SUMMARY lysoPCs with C20:4 and C18:2 were significantly restored by FCLM administration. PR:PROJECT_SUMMARY These metabolites may explain the above-mentioned differences in antiobesity PR:PROJECT_SUMMARY effects. Although only the changes in plasma lysophospholipids could not fully PR:PROJECT_SUMMARY explain antiobesity effects between non-fermented and fermented plant mixtures PR:PROJECT_SUMMARY from our results, we suggest that metabolomics approach could provide a way to PR:PROJECT_SUMMARY reveal the metabolite alterations in the complex fermentation process and PR:PROJECT_SUMMARY understand the differences or changes in bioactivity according to fermentation. PR:INSTITUTE Konkuk university PR:LAST_NAME Suh PR:FIRST_NAME Dong Ho PR:ADDRESS Neong-Dong-ro 120, Seoul, Kwang-Gin-gu, 05029, Korea, South PR:EMAIL sdh14031988@naver.com PR:PHONE 82-02-444-4290 #STUDY ST:STUDY_TITLE Comparison of Metabolites Variation and Antiobesity Effects of a Mixture of ST:STUDY_TITLE Cudrania tricuspidata, Lonicera caerulea, and the Soybean According to ST:STUDY_TITLE Fermentation in vitro and in vivo ST:STUDY_SUMMARY We used ultra-performance-liquid-chromatography with quadrupole-time-of-flight ST:STUDY_SUMMARY mass spectrometry to study the changes in metabolites in the mixture of Cudrania ST:STUDY_SUMMARY tricuspidata, Lonicera caerulea, and soybean (CLM) during fermentation. ST:STUDY_SUMMARY Additionally, the antiobesity effects of CLM and fermented-CLM (FCLM) were ST:STUDY_SUMMARY studied based on the analysis of plasma from high-fat diet (HFD)-fed mice. The ST:STUDY_SUMMARY levels of cyanidin and the glycosides of luteolin, quercetin, and cyanidin ST:STUDY_SUMMARY derived from L. caerulea were decreased, whereas the levels of luteolin and ST:STUDY_SUMMARY quercetin were increased during fermentation. Isoflavone glycosides and ST:STUDY_SUMMARY soyasaponins originating from the soybean were decreased, whereas their ST:STUDY_SUMMARY aglycones such as daidzein, glycitein, and genistein were increased. As for ST:STUDY_SUMMARY prenylated flavonoids from C. tricuspidata, these metabolites were decreased at ST:STUDY_SUMMARY the early stage of fermentation, and were increased at end of the fermentation. ST:STUDY_SUMMARY In terms of the functional food product, various metabolites derived from ST:STUDY_SUMMARY diverse natural products in CLM had complementary effects and demonstrated ST:STUDY_SUMMARY higher antioxidant and pancreatic lipase inhibition activities by fermentation; ST:STUDY_SUMMARY these activities were closely related to flavonoid aglycones including ST:STUDY_SUMMARY genistein, daidzein, glycitein, luteolin, and quercetin. In vivo experiment, ST:STUDY_SUMMARY several clinical parameters affected by HFD were remarkably improved by the ST:STUDY_SUMMARY administration of either CLM or FCLM, but there was a difference in the ST:STUDY_SUMMARY antiobesity effects. The levels of lysoPCs with C20:4, C16:0, and C22:6 were ST:STUDY_SUMMARY significantly attenuated by CLM administration, while the attenuated levels of ST:STUDY_SUMMARY lysoPCs with C20:4 and C18:2 were significantly restored by FCLM administration. ST:STUDY_SUMMARY These metabolites may explain the above-mentioned differences in antiobesity ST:STUDY_SUMMARY effects. Although only the changes in plasma lysophospholipids could not fully ST:STUDY_SUMMARY explain antiobesity effects between non-fermented and fermented plant mixtures ST:STUDY_SUMMARY from our results, we suggest that metabolomics approach could provide a way to ST:STUDY_SUMMARY reveal the metabolite alterations in the complex fermentation process and ST:STUDY_SUMMARY understand the differences or changes in bioactivity according to fermentation. ST:INSTITUTE Konkuk university ST:LAST_NAME Suh ST:FIRST_NAME Dong Ho ST:ADDRESS Neong-Dong-ro 120, Seoul, Kwang-Gin-gu, 05029, Korea, South ST:EMAIL sdh14031988@naver.com ST:PHONE 82-02-444-4290 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - ND1 Diet:ND SUBJECT_SAMPLE_FACTORS - ND2 Diet:ND SUBJECT_SAMPLE_FACTORS - ND3 Diet:ND SUBJECT_SAMPLE_FACTORS - ND4 Diet:ND SUBJECT_SAMPLE_FACTORS - ND5 Diet:ND SUBJECT_SAMPLE_FACTORS - ND6 Diet:ND SUBJECT_SAMPLE_FACTORS - ND7 Diet:ND SUBJECT_SAMPLE_FACTORS - ND8 Diet:ND SUBJECT_SAMPLE_FACTORS - ND9 Diet:ND SUBJECT_SAMPLE_FACTORS - ND10 Diet:ND SUBJECT_SAMPLE_FACTORS - HD1 Diet:HD SUBJECT_SAMPLE_FACTORS - HD2 Diet:HD SUBJECT_SAMPLE_FACTORS - HD3 Diet:HD SUBJECT_SAMPLE_FACTORS - HD4 Diet:HD SUBJECT_SAMPLE_FACTORS - HD5 Diet:HD SUBJECT_SAMPLE_FACTORS - HD6 Diet:HD SUBJECT_SAMPLE_FACTORS - HD7 Diet:HD SUBJECT_SAMPLE_FACTORS - HD8 Diet:HD SUBJECT_SAMPLE_FACTORS - HD9 Diet:HD SUBJECT_SAMPLE_FACTORS - HD10 Diet:HD SUBJECT_SAMPLE_FACTORS - HX1 Diet:HX SUBJECT_SAMPLE_FACTORS - HX2 Diet:HX SUBJECT_SAMPLE_FACTORS - HX3 Diet:HX SUBJECT_SAMPLE_FACTORS - HX4 Diet:HX SUBJECT_SAMPLE_FACTORS - HX5 Diet:HX SUBJECT_SAMPLE_FACTORS - HX6 Diet:HX SUBJECT_SAMPLE_FACTORS - HX7 Diet:HX SUBJECT_SAMPLE_FACTORS - HX8 Diet:HX SUBJECT_SAMPLE_FACTORS - HX9 Diet:HX SUBJECT_SAMPLE_FACTORS - HX10 Diet:HX SUBJECT_SAMPLE_FACTORS - HCLM1 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM2 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM3 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM4 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM5 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM6 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM7 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM8 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM9 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HCLM10 Diet:HCLM SUBJECT_SAMPLE_FACTORS - HFCLM1 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM2 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM3 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM4 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM5 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM6 Diet:HFCLM SUBJECT_SAMPLE_FACTORS - HFCLM7 Diet:HFCLM #COLLECTION CO:COLLECTION_SUMMARY Plasma 200uL were extracted with 1mL of 80% MeOH #TREATMENT TR:TREATMENT_SUMMARY Seven-week-old C57BL6J male mice were purchased from Daehan bio-link (Chungbuk, TR:TREATMENT_SUMMARY Republic of Korea). All mice were acclimated for 1 week under controlled TR:TREATMENT_SUMMARY conditions (temperature: 25 ± 2°C, relative humidity: 50 ± 5%, and 12 h TR:TREATMENT_SUMMARY light/dark cycle). The animals had free access to a normal diet (AIN-76A, TR:TREATMENT_SUMMARY Research Diets, Inc., NJ, USA) and water. After 1 week, the mice were randomly TR:TREATMENT_SUMMARY distributed into 5 groups: (1) group normal-diet (ND) (n = 11) was fed a normal TR:TREATMENT_SUMMARY diet for 6 weeks, (2) group HFD (named .D n = 10) was fed 60 kcal% fat (D1242, TR:TREATMENT_SUMMARY Research Diets, Inc., NJ, USA) for 6 weeks, (3) group HFD with xenical TR:TREATMENT_SUMMARY administration (HX, 50 mg?kg?1?day?1) n = 10, 6 weeks, (4) group HFD TR:TREATMENT_SUMMARY with CLM administration (HCLM, 2 g?kg?1?day?1) n = 10, 6 weeks; and (5) TR:TREATMENT_SUMMARY group HFD with FCLM (CLM fermented for 60 h) administration (HFCLM, 2 TR:TREATMENT_SUMMARY g?kg?1?day?1) n = 8, 6 weeks. In this study, we used xenical as a TR:TREATMENT_SUMMARY positive control for antiobesity effects. Xenical, CLM, and FCLM dissolved with TR:TREATMENT_SUMMARY saline were orally administered everyday into the stomach with an oral zonde TR:TREATMENT_SUMMARY needle. And, the equal volume of saline were applied for ND and HD groups. TR:TREATMENT_SUMMARY During the experimental periods, the feed intake and the body weight of the mice TR:TREATMENT_SUMMARY were measured daily. TR:TREATMENT_PROTOCOL_FILENAME DongHo_20160115_025706_PR_CO_The_animal_experiment.docx #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For blood collection, the mice were sacrificed by cardiac puncture. Plasma (100 SP:SAMPLEPREP_SUMMARY µL) was subjected to extraction with cold methanol (500 µL) on a MM400 mixer SP:SAMPLEPREP_SUMMARY mill (Retsch®, Haan, Germany) with the frequency 30 s?1 for 5 min. The SP:SAMPLEPREP_SUMMARY suspension was centrifuged at 12,578 g for 10 min at 4°C. The supernatant was SP:SAMPLEPREP_SUMMARY filtered through a 0.2-µm PTEE filter and evaporated in a speed vacuum SP:SAMPLEPREP_SUMMARY concentrator. The final concentration of the plasma was 5 mg/mL in methanol for SP:SAMPLEPREP_SUMMARY the UPLC-Q-TOF-MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (100 x 2mm, 1.7um) CH:COLUMN_NAME Corp.) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Waters Micromass QTOF Premier MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples ND1 ND2 ND3 ND4 ND5 ND6 ND7 ND8 ND9 ND10 HD1 HD2 HD3 HD4 HD5 HD6 HD7 HD8 HD9 HD10 HX1 HX2 HX3 HX4 HX5 HX6 HX7 HX8 HX9 HX10 HCLM1 HCLM2 HCLM3 HCLM4 HCLM5 HCLM6 HCLM7 HCLM8 HCLM9 HCLM10 HFCLM1 HFCLM2 HFCLM3 HFCLM4 HFCLM5 HFCLM6 HFCLM7 Factors Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:ND Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HD Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HX Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HCLM Diet:HFCLM Diet:HFCLM Diet:HFCLM Diet:HFCLM Diet:HFCLM Diet:HFCLM Diet:HFCLM LysoPC(14:0) 174 185 166 163 164 213 158 171 212 176 184 160 164 146 181 154 173 163 176 174 174 167 255 167 217 180 287 207 180 217 151 132 120 122 135 135 128 150 137 134 128 162 135 128 151 172 142 LysoPC(18:0)_1 3053 2525 2973 2878 3274 2888 3340 3499 3431 3975 4168 4296 4696 4192 4074 3604 4493 3959 4261 4670 4800 3719 3967 2896 3651 3231 3294 4239 4314 4067 3983 3507 4195 4154 4270 4075 4656 4598 4822 4470 5344 4331 5818 5487 4464 4279 4217 LysoPC(18:0)_2 10662 10382 11113 10201 10723 9872 10212 10714 10177 10151 10756 12092 12028 10548 11025 12864 14181 11739 12786 13829 11968 11272 12949 8809 11231 9732 11497 12514 11445 11495 11062 11360 12074 14271 12011 11950 13420 12809 14886 12042 14244 13147 14910 13446 12963 13137 12048 LysoPC(18:1) 9296 9825 10043 9494 9706 10290 10589 9397 11179 9774 10310 9372 10298 8520 9167 9158 9301 8905 9394 9598 10064 9291 9971 9613 10493 9524 10677 10251 9482 10099 8310 9842 8687 9275 9279 7216 8512 8495 8047 8519 8830 9465 8417 8644 8675 9112 9103 LysoPC(18:2)_1 3974 3352 4067 4034 3709 4427 4866 4472 4178 4553 3724 3185 3828 3560 4088 3017 2952 3405 3447 3248 4799 3850 3395 4603 4662 3737 3773 4224 3691 4462 3540 3769 3296 3330 3858 3281 3799 3748 3039 3751 3412 3694 3854 4269 3710 4128 4538 LysoPC(18:2)_2 18182 16858 17453 18542 17199 16563 15964 16165 16672 17154 12808 13314 12060 12363 13154 13501 12663 13300 13858 13014 15396 15970 14857 17995 17532 16324 14947 17155 13657 15791 12395 13153 13037 11858 15272 11718 14912 13697 12147 13193 12921 12209 13088 15033 11650 15825 17514 LysoPC(18:3)_1 223 226 164 194 171 228 167 176 239 223 103 124 111 107 105 113 78 100 111 99 135 100 135 196 170 135 149 154 103 165 104 97 80 77 87 93 91 109 77 80 85 95 87 102 89 113 111 LysoPC(18:3)_2 381 355 295 404 278 386 332 276 296 381 241 254 226 208 289 193 162 200 209 233 283 275 309 376 278 290 296 208 211 425 228 276 189 163 197 191 228 236 228 263 146 226 177 279 193 379 314 LysoPC(20:2) 456 503 594 424 495 610 711 596 835 586 540 532 601 495 460 397 432 353 402 371 732 695 567 778 746 782 789 751 688 739 308 493 311 322 377 283 313 309 266 276 343 350 334 297 393 417 454 LysoPC(20:3)_1 228 235 397 207 297 320 683 410 437 330 439 354 497 442 420 210 313 172 262 232 396 181 231 255 231 193 210 284 254 293 188 315 203 259 243 129 138 152 156 164 210 243 167 207 301 141 158 LysoPC(20:3)_2 1548 1874 2393 1535 1859 1832 2843 2399 2473 1822 2304 2067 2558 2221 2081 1517 1970 956 1578 1577 2157 1238 1397 1691 1488 1379 1342 1656 1438 1527 915 1688 1135 1249 1231 682 780 684 825 763 967 1115 785 955 1367 767 782 LysoPC(20:4)_1 1574 1454 1970 1688 1914 1725 2317 2047 1736 1683 2710 2748 3090 2698 2862 2085 1955 1503 2218 2171 2750 1782 2073 2350 1982 1929 2420 2219 2210 2563 2184 3128 2004 2260 1832 1475 1805 2002 1784 2048 1930 2828 1969 2138 2820 1760 1818 LysoPC(20:4)_2 5604 6070 6198 6778 6334 5881 6685 6749 5020 4779 9228 8566 9911 8360 8380 8578 7273 5238 7323 7796 7747 6935 7549 7776 6633 6910 7696 7388 7086 8495 6373 9079 7499 8051 6065 5282 6947 6901 6677 6869 5849 8120 6566 6867 8437 6303 5622 LysoPC(22:5) 358 306 398 314 306 321 494 344 347 314 471 386 536 484 511 302 300 291 339 385 437 253 281 319 274 271 226 250 265 265 272 510 273 278 310 181 243 278 214 234 356 417 317 376 423 262 249 LysoPC(22:6)_1 1183 1116 1411 1335 1373 1231 1765 1328 1401 1271 2340 1972 2628 2216 2454 1641 1629 1453 1893 1814 2076 1171 1489 1481 1301 1204 1486 1381 1581 1613 1718 2411 1653 1673 1612 1020 1491 1785 1580 1379 1851 2476 1926 2115 2511 1453 1397 LysoPC(22:6)_2 6168 6108 6339 6446 6443 5554 6548 5795 5683 5799 9054 8123 10495 8966 8477 8240 7451 6858 7961 9600 8266 5993 6871 7154 6662 6877 6659 5737 7513 7338 7494 9500 7543 8019 7202 4897 6040 7619 6964 6539 8542 9540 7541 8451 10801 6921 5714 LysoPE(18:0) 422 378 321 369 362 384 264 319 353 363 197 193 200 169 192 204 217 213 225 217 176 257 247 209 286 252 269 245 246 278 171 172 161 169 203 189 159 207 187 188 173 187 184 164 159 255 221 LysoPE(22:6)_1 131 106 140 131 114 150 149 149 183 158 256 196 247 154 222 140 184 205 172 204 174 159 194 151 181 145 171 158 225 190 220 170 146 169 156 88 176 225 184 204 264 254 221 229 246 140 130 LysoPE(22:6)_2 1312 987 1033 1118 885 1195 987 1230 1322 1194 1762 1398 1768 1142 1410 1178 1535 1566 1328 1626 1252 1337 1552 1184 1288 1171 1264 1397 1649 1358 1459 1147 1046 1292 1190 721 1381 1774 1559 1415 1895 1759 1664 1601 1638 1073 922 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB_ID LysoPC(14:0) HMDB10379 LysoPC(18:0)_1 HMDB10384 LysoPC(18:0)_2 HMDB10384 LysoPC(18:1) HMDB02815 LysoPC(18:2)_1 HMDB10386 LysoPC(18:2)_2 HMDB10386 LysoPC(18:3)_1 HMDB10387 LysoPC(18:3)_2 HMDB10387 LysoPC(20:2) HMDB10392 LysoPC(20:3)_1 HMDB10393 LysoPC(20:3)_2 HMDB10393 LysoPC(20:4)_1 HMDB10395 LysoPC(20:4)_2 HMDB10395 LysoPC(22:5) HMDB10402 LysoPC(22:6)_1 HMDB10404 LysoPC(22:6)_2 HMDB10404 LysoPE(18:0) HMDB11129 LysoPE(22:6)_1 HMDB11526 LysoPE(22:6)_2 HMDB11526 METABOLITES_END #END