Novel Tocotrienols Of Rice Bran Modulate Cardiovascular-Books Pdf

Novel tocotrienols of rice bran modulate cardiovascular
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Novel tocotrienols reduce cardiovascular risk Qureshi et al. Various isomeric forms of tocotrienols are distinguished by terol and LDL cholesterol levels by 16 to 20 in respond. the number and location of methyl groups on the chroman ing individuals 35 out of 47 hypercholesterolemic. rings The Y B y and tocotrienols are Y 5 7 subjects All the subjects in this trial were allowed to. X trimethyl B 5 8dimethyl y 7 dimethyl and 6 consume their normal diets free living Because of the. 8 monomethyl respectively depending on the number large number of nonrespondants these effects were not. and position of the methyl substituents on the benzene ring statistically significant for the entire group Therefore the. of the chroman moiety l4 The two novel tocotrienols iso present double blind study was initiated to measure the. lated from specially processed rice bran eluting at 21 min effects of TRF on hypercholesterolemic subjects restricted. peak 21 P and 25 min peak 25 P on high perfor to a controlled diet NCEP Step l diet. mance chromatography HPLC system have been charac, terized as desmethyl and didesmethyl no methyl groups on. the benzene ring of the chroman moiety tocotrienols 5 The Methods and materials. unsaturated tocotrienols unlike saturated tocopherols ex. hibit varying degrees of cholesterol lowering activity de TRF25. pending on the isomer i e B OL y 6 y Oryzanol free TRF was prepared by molecular distillation Dr. desmethyl didesmethyl The tocotrienols inhibit Laxman Singh Vitamins Inc Chicago IL USA of the oil. cholesterol synthesis by suppressing B hydroxy B meth extracted from rice bran which was processed and supplied by. Bionutrics Inc Phoenix AZ USA The TRF2 consisted of 6. ylglutaryl coenzyme A HMG CoA reductase activity cw tocopherol 12 5 cu tocotrienol 21 y tocotrienol 10 to. through two post transcriptional actions increasing the cotrienol 4 5 d tocotrienol 17 P ocotrienol d di desmethyl. controlled degradation of reductase protein and decreas tocotrienol 3 4 dihydro 2 4 8 12 tnmethyltrideca 3 E 7 E. ing the efficiency of the translation of HMG CoA reduc I 1 trienyl 2 H I benzopyran 6 o 18 unidentified tocotrien. tase messenger RNA mRNA ols and 10 sterols and triglycerides. The commercially stabilized rice bran and rice bran oil. contains 125 to 830 pg g tocols Rice bran constitutes about Study population. 10 of rough rice grain and contains 18 to 22 oil Milling. Adults enrolling in this project were recruited from a hypercho. activates an endogenous lipase activity resulting in the rapid. lesterolemic population screened at the University of Illinois. deterioration of the oil rendering it unsuitable for human. Chicago Prospective participants were grouped according to. consumption The temperature sensitive lipase is inacti cholesterol level median and subgrouped by sex The members. vated by a two step process a 3 second exposure to 135 C of each subgroupwere randomizedinto two groups using a. followed by 3 min exposure to 100 C This processing randomnumbertable There were 21 participantsin the placebo. stabilizes the rice bran for use in high fiber cereal prod group 9 males and 12 females 42 8 t 14 0 years of age 27 3 5. ucts Stabilizing the rice bran at high temperature 5 0 body mass index and 6 75 1 12 mmol cholesterol L There. 180 C under vacuum 2 psi for 60 min before extraction were 20 participants in TRF group 10 males and 10 females. increases by 2 to 3 fold the quantities of the known 41 5 15 5 years of age 25 5 body mass index and 6 57 2 0 77. tocotrienols and also produces six novel analogs of toco mmol cholesterol L There were 16 female participants over 50. trienols 20 One of these novel analogs is didesmethyl years old eight in Placebo and eight in TRF groups and the. remaining six were 33 to 37 years old four in placebo and two in. tocotrienol P no methyl groups on the chroman rings in TRFzs groups Exclusion criteria included weight 125 of. the oil the proportion of which has been shown to increase Metropolitan Life relative weights use of cholesterol altering. with such processing to 20 to 30 of the total tocols medication an elevated serum glutamate pyruvate or glutamate. Molecular distillation of the oil yields four fractions the oxaloacetate transaminase activity an elevated blood urea nitrogen. first consisting of free fatty acids and the y oryzanols The or glucose value diabetes or a liver renal or hypertensive. second the novel tocotrienol enriched fraction TRF disease All participants signed an informed consent statement. consists of the Pzs tocol complex 90 with small quanti which was approved by Institutional Review Board of the Medical. ties of sterols and triglycerides Animal trials show P2s Center of University of Illinois Chicago. tocotrienol to be the most efficacious mevalonate suppres. sive cholesterol lowering tocotrienol isomer 15 Experimental design. As suggested in the foregoing discussion the hypocho The experiment consisted of three phases the first an alcohol free. lesterolemic action of tocotrienols affects endogenously free choice diet phase baseline for a period of 4 weeks followed. synthesized cholesterol TRF also provides antithrombotic by a second phase 4 weeks during which all participants were. and antioxidant activities is Estimates of cardiovascular counseled to follow the National Cholesterol Education Program. disease risk are nominally assessed by measures of serum NCEP Step l diet The participants were continued on the NCEP. total cholesterol low density lipoprotein LDL cholesterol Step l diet during the third 4 week phase treatment During the. high density lipoprotein HDL cholesterol as well as treatment phase participants assigned to the placebo group re. apolipoproteins A I apo A I apo B Lipoprotein a ceived four capsules each containing 300 mg corn oil tocopherol. Lp a thromboxane B2 TxB2 and platelet factor 4 stripped corn oil was used to prepare these capsules Tekled Test. Diets Madison WI USA Participants assigned to the TRF. PF4 These considerations led us to test the effect of group received four capsules each containing 50 mg TRF and. molecularly distilled y oryzanol free TRF on the cardio 250 mg corn oil In total the TRF capsules supplied daily 25. vascular risk profile of hypercholesterolemic human sub mg a tocotrienol 42 mg y tocotrienol 20 mg S tocotrienol 34 mg. jects P tocotrienol 9 mg d tocotrienol 22 12 mg o tocopherol and. In an earlier experiment supplementation with tocotrien 36 mg unidentified tocols The participants recorded their daily. 01s Palmvitee from palm oil lowered serum total choles food intakes during the final 3 days of each phase Participants. J Nutr Biochem 1997 vol 8 May 291, Research Communications. were contacted by telephone during each phase for an unantici determined by using a Shimadzu Model C R3A integrator Shi. pated 24 hr recall of food intake Diet records and 24 hr recalls madzu Wood Dale IL USA A 10 micron 30 cm X 4 0 mm i d. were analyzed Nutrition Coordinating Center University of normal phase silica column Waters Associates Milford MA. Minnesota Minneapolis MN USA if required a participant was USA was used to separate various tocols The eluting solvent was. individually counseled to modify food intake to meet the goals of 0 5 vol vol isopropyl alcohol in hexane 25 Retention time of the. the NCEP Step l diet or to maintain weight individual peaks of the unknown tocols were compared against the. Initial measures included the participants height weight blood retention time of the pure standard tocols The tocols were eluted. pressure history of significant diseases medications and alcohol under these conditions in the sequence a tocopherol 4 6 min. use Weights were recorded weekly Venous blood samples 12 hr cw tocotrienol 5 3 min B tocopherol 6 4 min y tocopherol 7 4. fast 7 00 9 00 am were drawn at screening The participants min B tocotrienol 8 6 min y tocotrienol 9 4 min S tocoph. were screened 1 to 3 weeks before obtaining the baseline values erol 14 6 min S tocotrienol 16 2 min desmethyl tocotrienol. At screening the participants were counseled to follow their 21 2 min P2 and didesmethyl tocotrienol 25 3 min P J as. normal dietary patterns exclusive of alcohol consumption Venous reported earlier from cereals and serum samples 14 15 21 25 1i5 The. blood samples were drawn at the termination of the baseline phase pure tocols tocopherols and tocotrienols were provided by Dr B. at week 4 followed by NCEP Step l diet weeks 7 and 8 and Pearce of Bristol Myers Squibb Pharmaceutical Research Institute. treatment weeks 11 and 12 phases Processed samples were Wallingford CT USA. coded and held at 72 C until analysis following the Treatment. Statistical analyses, Analyses The data were analyzed by using the GLM procedure of SAS. Statistical Analysis System for personal computers to test the. Plasma and serum were frozen and shipped on dry ice to Advanced study hypothesis Duncans multiple range test was used to test. Medical Research Madison WI USA The analyses of the coded whether the treated groups differed from the placebo for serum. samples were performed at Advanced Medical Research Serum lipid parameters Repeated measures two way ANOVA was used. triglycerides glucose and cholesterol were estimated manually to test whether changes in serum lipid parameters occur in the. incubation time was 10 min for each estimation of lipid param course of supplementation and whether there were between and. eters with reagent kits from Sigma Chemical Co St Louis MO within subjects differences because all observations were re. USA LDL cholesterol were precipitated from 200 pL of serum quired available degrees of freedom were reduced by this statis. with 25 PL of a mixture of 9 7 mM phosphotungstic acid and 0 4 tical approach The treatment effects on cholesterol were also. M MgCl The preparation was mixed for 10 min at room evaluated using the paired two tailed t test StatView Abacus. temperature and then centrifuged at 12 000 X g for 10 min The Concepts Berkeley CA USA Data are reported as mean t SD. supematant fraction was decanted and analyzed for HDL choles in the text The statistical significance level was set at 5. terol The precipitate was dissolved in 200 PL of 0 1 M sodium. citrate and the concentration of LDL cholesterol could be deter. mined The LDL cholesterol was estimated by Friedewald s for. mula by subtracting the total cholesterol from HDL choles Results. terol triglycerides 5 Serum apo A I apo B Sigma Chemical The intervention for all subjectsduring the last two phases. Co St Louis MO USA platelet factor 4 PF4 Abbott Labora weeks 5 to 12 was the NCEP Step l diet The analysisof. tories Chicago IL USA and thromboxane B TxB Chemicon. International El Segundo CA USA concentrations were deter all the subjects during this phase reveals that the subjects. mined by radio immunoassay kits Lp a was assayed with an significantly decreased their reported intakes of energy. immunoassay kit from Terumo Medical Corporation Diagnostic 23 P O Ol carbohydrate 19 P 0 05 fat. Division Elkton MD USA 40 P 0 03 saturatedfatty acids 38 P O Ol. Sera were processed for the determination of tocols Serum 1 O and cholesterol 33 P 0 04 as shown in Table 1 The. ml was transferred to a 20 mL culture tube 8 mL hexane was detailed dietary intake of protein fat carbohydrate choles. added mixed for 20 min centrifuged for 10 min at 850 X g and terol fiber and alcohol at various phaseshas been outlined. the hexane layer was transferred to a 15mL conical tube The in Table 1 Moreover the body weight in Placebo and. hexane extraction was repeated the hexane extracts containing the Treatment groups showed no significant difference during. tocols were combined and evaporated under vacuum at 40 C The NCEP Step l diet and NCEP Step l diet TRF as. aqueous layer was taken to dryness in a vacuum oven 2 psi and compared to Baseline Table I The sole restriction during. held at 180 C for I hr The serum residue was then extracted with. 8 mL methanol The methanol was removed under vacuum to the acclimation phase that of alcohol abstention had little. measure the tocols bound to the protein Tocols extracted by each effect as shown by week 4 Baseline values for all. procedure were dissolved in 200 p L hexane transferred to parametersfrom Screening values Tables 2 4 The dietary. injecting vials the vials were capped and centrifuged for 4 min restriction testsat weeks 7 and 8 NCEP Step l diet alone. The hexane and methanol soluble tocols present in the HDL and lowered serum total cholesterol and LDL cholesterol and. LDL separated by phosphotungstic acid and MgCl fractions apo B 5 8 and 8 NS respectively from Baseline. were similarly extracted The clear solutions were analyzed by slight levels Table 2 TRF supplement tests at weeks 11 and. modification of the well established HPLC method of Hakkarainen et 12 decreasedtotal cholesterol LDL cholesterol and apo B. 16 23 and 15 P 0 05 respectively from, The HPLC system consisted of a continuous flow 307 pump Baseline values There were further decreasesin the Placebo. Gilson Madison WI USA flow rate 1 3 mUmin pressure 0 4. psi and a Gilson s Model 231 401 auto sampler loop size 20 group of 2 3 and 3 P O OS respectively in. pL a Shimadzu Model RF 535 fluorescence monitor set at an these parameters caused by the NCEP Step l diet at. excitation wavelength of 295 nm and an emission wavelength of 12 week period Table 2. 330 nm and for UV monitoring Shimadzu UV VIS SPD IO AV The dietary modification of these subjects did not pro. This research was supported by a grant from Bionutrics Inc Phoenix Arizona The didesmethyl tocotrienol P amp is a late eluting component present in the tocotrienol rich fraction TRF of oil from processed rice bran the 25 min HPLC fraction The designation TRF is used to distinguish this

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