Seed Kernel Oil PUFA Lipid-lowering Activity And Oxidative Damage On Research Pepper Based 3T3-L1 Cells

The prickly ash seeds were often burnt or send to landfill as wastes in prickly ashprocessing production. Whereas the prickly ash seeds the theoretical yield of theprickly ash seeds was up to600000tons, which was20%higher than that of peel field,then that caused great environmental pollution. Studies had shown that the prickly ashseeds, which contain abundant polyunsaturated fatty acids and proteins, be regardedas waste had caused a big waste of useful resources. How to explore and use theseprickly ash seeds more efficiently was of common concern by many fields experts.Our laboratory extracted pure natural and high nutrition edible oil, the pricklyash seed kernel oil through the spiral squeezing mechanism in the prickly ash seeddepth development. The study also found that the prickly ash seed kernel oil containrich polyunsaturated fatty acids(PUFA), especially alpha linolenic of n-3series andlinoleic of n–6series were34.0%and29.9%, and the proportion was appropriateabout1:1. Our laboratory extracted the prickly ash seed kernel oil polyunsaturatedfatty acids ethyl ester that the proportion of alpha linolenic acids ethyl ester of n-3series and linoleic acids ethyl ester of n–6series was about5:1by silver nitratecomplexiation and the sodium ethylate as catalyst.On this basis, this paper studied thelipid-lowing active of the prickly ash seed kernel oil PUFA and prickly ash seedkernel oil PUFAEE. Firstly, the high purity of alpha linolenic acid and linoleic acidwere studied on3T3-L1cell proliferation and differentiation. Then the prickly ashkernel oil PUFA and prickly ash seed kernel oil PUFAEE were studied on theinfluence of3T3-L1cell proliferation and differentiation, and tried to study themechanism on the PUFA influencing the differentiation of3T3-L1cell through theaspect of the lipid peroxide oxidation damage.(1) The study about n-3/n-6PUFA influenced the proliferation anddifferentiation of3T3-L1cell3T3-L1preadipocytes were used as target cells to explore the effect of differentconcentration ALA and LA on their proliferation measured by MTS assay and cellmorphology analysis.3T3-L1cells under the induction of dexamethasone, insulin and xanthine may specificity become into mature fat cells, which could be dye by theoil red O. The quantity of oil red O could indirectly response the influence of differentconcentration ALA, LA,1:1ALA/LA on the3T3-L1cell differentiation. MTS resultsshowed that ALA and LA inhibited significantly proliferation of3T3-L1preadipocytes in dose-dependent and time-dependent manner at a certainconcentration range, and ALA inhibited proliferation more efficient than LA; Cellmorphology analysis results showed that ALA and LA were cytotoxic to3T3-L1preadipocytes at a high concentration. The inverted electron microscope observingand oil red staining and detection results showed that ALA, LA and1:1ALA/LAmixed fatty acid inhibited significantly differentiation of3T3-L1preadipocytes indose-dependent manner at range of12.5~200μM, and ALA inhibited differentiationmore efficient than LA, and the1:1ALA/LA mixed fatty acid inhibited differentiationof3T3-L1preadipocytes more efficient at the same concentration above100μM thanALA.(2) The study about the effects of the prickly ash seed kernel oil PUFA andprickly ash kernel oil PUFAEE on the3T3-L1cell proliferation and differentiationOn the basis of previous research, this research adopted100μM,200μM,400μMconcentrations of the prickly ash seed kernel oil PUFA and prickly ash seed kernel oilPUFAEE to intervene3T3-L1cell proliferation process, and tested the relativenumber of living cells by MTS to indirectly response the inhibition effect toproliferation of3T3-L1cell, and detect the cytotoxic effect in the high concentrationby combining the cell morphological analysis.3T3-L1cells were inducted throughtraditional cocktail and were dyed by oil red O, then detected the effect of the pricklyash seed kernel oil PUFA and prickly ash seed kernel oil PUFAEE of200μM onthe3T3-L1differentiation by colorimetric analysis method. MTS results showed thatthe prickly ash seed kernel oil PUFA and prickly ash seed kernel oil PUFAEEinhibited significantly proliferation of3T3-L1preadipocytes in dose-dependent andtime-dependent manner at a certain concentration range; Cell morphology analysis results showed that the prickly ash seed kernel oil PUFA was cytotoxic to3T3-L1preadipocytes at concentration of400μM, however the prickly ash seed kernel oilPUFAEE was not. The results of differentiation experiments showed that the pricklyash seed kernel oil PUFA and prickly ash seed kernel oil PUFAEE inhibitedsignificantly differentiation of3T3-L1preadipocytes, and the prickly ash seed kerneloil PUFAEE inhibited differentiation more efficient than the prickly ash seed kerneloil PUFA.(3) The study about the effects of oxidative damage of the prickly ash seedkernel oil PUFA and prickly ash seed kernel oil PUFAEE to3T3-L1celldifferentiationThe effects of oxidative damage of the prickly ash seed kernel oil PUFA andprickly ash seed kernel oil PUFAEE to3T3-L1cell differentiation were studiedthrough detect the level of MDA, GSH and SOD in the cells. The results showed thatthe levels of MDA under the intervention of the prickly ash seed kernel oil PUFA andprickly ash seed kernel oil PUFAEE were more significantly higher than thecorresponding control group, however the levels of SOD and GSH were significantlylowered. The prickly ash seed kernel oil PUFAEE has lower levels of MDA andhigher levels of SOD and GSH than the group of the prickly ash seed kernel oil PUFAbecause of better oxidation stability. The results point out that the more efficientinhibitory of differentiation of PUFAEE on the same of concentration may be relatedto its better oxidation stability.