Journal List > J Nutr Health > v.51(4) > 1100548

J Nutr Health. 2018 Aug;51(4):275-286. Korean.
Published online August 31, 2018.  https://doi.org/10.4163/jnh.2018.51.4.275
© 2018 The Korean Nutrition Society
Metabolites profiling and hypolipidemic/hypocholesterolemic effects of persimmon (Diosyros kaki Thumb.) by different processing procedures: in vitro and in vivo studies
Soo-Yeon Park,1,** Eun-Kyung Oh,1,** Yeni Lim,1 Ji-Yoon Shin,2 Hee-Ah Jung,3 Song-Yi Park,3 Jin Hee Lee,3 Jeong-Sook Choe,4 and Oran Kwon1,2
1Department of Nutritional Science and Food Management, Ewha Womans University, Seoul 03760, Korea.
2Ewha Graduate School of Converging Clinical&Public Health, Seoul 03760, Korea.
3Department of Food Science and Biotechnology, CHA University, Pocheon, Gyeonggi 11160, Korea.
4Department of Agrofood Resources, Rural Development Administration National Institute of Agricultural Sciences, Jeonju, Jeonbuk 55365, Korea.

To whom correspondence should be addressed. tel: +82-2-3277-6860, Email: orank@ewha.ac.kr

**Equally contributed to this work as the first author

Received June 18, 2018; Revised July 06, 2018; Accepted July 09, 2018.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


Abstract

Purpose

Our previous study demonstrated that persimmon (Diospyros kaki Thumb.) at different stages of ripening provided different protective effects against high-fat/cholesterol diet (HFD)-induced dyslipidemia in rats. In this study, we compared the metabolites profile and gene expressions related to triglyceride (TG)/cholesterol metabolism in vitro and in vivo after treating with persimmon water extracts (PWE) or tannin-enriched persimmon concentrate (TEP).

Methods

Primary and secondary metabolites in test materials were determined by GC-TOF/MS, UHPLC-LTQ-ESI-IT-MS/MS, and UPLC-Q-TOF-MS. The expression of genes related to TG and cholesterol metabolism were determined by RT-PCR both in HepG2 cells stimulated by oleic acid/palmitic acid and in liver tissues obtained from Wistar rats fed with HFD and PWE at 0, 150, 300, and 600 mg/d (experiment I) or TEP at 0, 7, 14, and 28 mg/d (experiment II) by oral gavage for 9 weeks.

Results

PLS-DA analysis and heatmap analysis demonstrated significantly differential profiling of metabolites of PWE and TEP according to processing of persimmon powder. In vitro, TEP showed similar hypolipidemic effects as PWE, but significantly enhanced hypocholesterolemic effects compared to PWE in sterol regulatory element-binding protein 2 (SREBP2), HMG-CoA reductase (HMGCR), proprotein convertase subtilisin/kexin type 9 (PCSK9), cholesterol 7α-hydroxylase (CYP7A1), and low density lipoprotein receptor (LDLR) gene expression. Consistently, TEP and PWE showed similar hypolipidemic capacity in vivo, but significantly enhanced hypocholesterolemic capacity in terms of SREBP2, HMGCR, and bile salt export pump (BSEP) gene expression.

Conclusion

These results suggest that column extraction after hot water extraction may be a good strategy to enhance tannins and long-chain fatty acid amides, which might cause stimulation of hypocholesterolemic actions through downregulation of cholesterol biosynthesis gene expression and upregulation of LDL receptor gene expression.

Keywords: Diospyros kaki Thumb.; metabolites profiling; Hep G2 cells; diet-induced hyperlipidemic rats

Figures


Fig. 1
Partial lest-squares discrimination analysis (PLS-DA) score plots using the GC-TOF-MS (A) and UHPLC-LTQ-ESI-IT-MS/MS/UPLC-Q-TOF-MS (B) data sets and heat map of specific metabolites in PDP, PWE, and TEP (C). PDP, persimmon dried powder; PWE, persimmon water extract; TEP, tannin-enriched persimmon concentrate. R2X and R2Y were the cumulative modeled variation in X and Y matrix, respectively. Q2 were the cumulative predicted variation in Y matrix.
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Fig. 2
Effects of PWE and TEP on intracellular lipid accumulation (A and B, respectively) and FAS gene expression (C) in HepG2 cells stimulated by free fatty acids (0.66 mM oleic acid + 0.33 mM palmitic acid). PWE, persimmon water extract; TEP, tannin-enriched persimmon concentrate. HepG2 cells were grown in serum-free medium overnight and incubated in 1mM OA/PA-BSA complex in the absence or presence of PWE (A) or TEP (B) for an additional 24h. Values are means from three independent experiments conducted with triplicate treatments, with standard error represented by vertical bars. *Significantly different at p < 0.05 by Dunnett's test compared with FFA loading.
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Fig. 3
Effects of PWE and TEP on cholesterol synthesis (A) and cholesterol metabolism (B) in HepG2 cells stimulated by free fatty acids (0.66 mM oleic acid + 0.33 mM palmitic acid). PWE, persimmon water extract; TEP, tannin-enriched persimmon concentrate. HepG2 cells were grown in serum-free medium overnight and incubated in 1mM OA/PA-BSA complex in the absence or presence of PWE (A) or TEP (B) for an additional 24h. Values are means from three independent experiments conducted with triplicate treatments, with standard error represented by vertical bars. *Significantly different at p < 0.05 by Dunnett's test compared with FFA loading.
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Fig. 4
Effects of PWE (A) and TEP (B) on hepatic lipogenesis (SREBP1c and FAS) in rats fed a high-fat/cholesterol diet. PWE, persimmon water extract; TEP, tannin-enriched persimmon concentrate, HF, high-fat/cholesterol control; HL, HF + low-dose PWE/TEP; HM, HF + middle-dose PWE/TEP; HH, HF + high-dose PWE/TEP. Values are expressed as mean ± SE (n = 10 for each group). *Significantly different at p < 0.05 by Dunnett's test compared with HF group.
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Fig. 5
Effects of PWE (A) and TEP (B) on hepatic cholesterol metabolism (SREBP2, LDLR, HMGCR and CYP7A1, BSEP) in rats fed a high-fat/cholesterol diet. PWE, persimmon water extract; TEP, tannin-enriched persimmon concentrate, HF, high-fat/cholesterol control; HL, HF + low-dose PWE/TEP; HM, HF + middle-dose PWE/TEP; HH, HF + high-dose PWE/TEP. Values are expressed as mean ± SE (n = 10 for each group). *Significantly different at p < 0.05 by Dunnett's test compared with HF group.
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Fig. 6
Overview diagram of research scope and flow
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Tables


Table 1
Primer sequences used for real-time PCR
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Notes

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01169502)” Rural Development Administration, Republic of Korea.

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