Obesity has become an important global public health problem.Natural products have always been an important direction of obesity prevention research.Pine pollen is a kind of traditional Chinese medicinal and food material, which is rich in phenolics and have activities for lowering lipids and improving lipid metabolism in vivo.However, previous studies have found that most phenolic ingredients in pine pollen exist in the form of binding state, which makes it unable to fully exert its activity of lowering lipids.In order to increase the lipid-reducing effect of pine pollen, fermentation by edible fungus Rhizopus oryzae was employed.Clean and dry pine pollen was used as the sole source of nutrients (substrate) for liquid medium (solid-liquid-ratio 1∶20) and was fermented in flask with 10% (mL∶g) inoculum (containing 106 spores/mL, 30 ℃, 180 r/min for 4 days.The effects of unfermented and fermented pine pollen (NFPP, FPP) on 3T3-L1 differentiation and lipid metabolism, including differentiation factors CCAAT enhancer-binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), liposynthetic factors FAS, lipolysis factors hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL) and fatty acid oxidation factor peroxisome proliferator-activated receptor alpha (PPARα), further the related mechanism was investigated.The results of HPLC showed that compared with NFPP, the contents of protocatechuic acid, p-courmaric acid, taxifolin and naringenin were increased by 176.30%, 10.42%, 88.43%, and 326.75%, respectively, while catechin was decreased by 32.51%.The total levels of five phenolic ingredients in pine pollen increased significantly, from 2 091.63 μg/g to 3 686.62 μg/g, and naringenin (NAR) increased from 475.06 μg/g to 2 027.30 μg/g.Oil red O staining and quantitative results showed that FPP could significantly reduce the lipid accumulation in 3T3-L1 cells, which was better than NFPP under the same concentration and at 400 μg/mL, the lipid accumulation of FPP was only 57.59%.In addition, the results of q-PCR and Western blot showed that FPP was able to significantly inhibit the adipocyte differentiation factor C/EBPα and PPARγ, up-regulate lipolysis-related factors HSL, ATGL and promote the expression of fatty acid oxidation factor PPARα in a dose-dependent manner (P<0.05).At 400 μg/mL, the mRNA expression of C/EBPα, PPARγ, HSL, ATGL, and PPARα in FPP were 0.25, 0.38, 6.37, 7.91 and 10.89 times of the blank control group, respectively, compared with NFPP.The results of western blot tests were almost consistent with q-PCR and except for the low concentration group of FPP (200 μg/mL), the other sample groups had almost no impact on FAS.It may due to adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) activation (upregulated ratio of p-AMPKα /AMPKα, p-AMPKβ1/ AMPKβ1 and p-ACC/ACC).The significant inhibited expression of PPARγ and C/EBPα, increased expression of PPARα and ATGL induced by naringenin may be mainly responsible for the lipid-reducing effects of FPP.In conclusion, FPP has a better effect on reducing lipid accumulation in 3T3-L1 cells, which may be mainly attributed to the increase of phenolic ingredients, especially naringenin.This research may provide theoretical guidance for the development of new pine pollen product with higher lipid-reducing activity.
WANG Xuan
,
LIU Qisong
,
GAO Fukai
,
HE Zhiyong
,
QIN Fang
,
WANG Zhaojun
,
CHEN Qiuming
,
CHEN Jie
,
ZENG Maomao
. Fermentation of pine pollen by Rhizopus oryzae and its effect on adipocyte differentiation and lipid metabolism of 3T3-L1 cells[J]. Food and Fermentation Industries, 2023
, 49(12)
: 1
-9
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DOI: 10.13995/j.cnki.11-1802/ts.033779
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