Dietary coconut water vinegar for improvement of obesity-associated inflammation in high-fat-diet-treated mice

Figures & data

Table 1. Grouping of the high-fat-diet (HFD) induced obese mice according to treatment.

Group	Treatment
UT (untreated obese control)	Induced obese mice given HFD + distilled water (control)
CL (low-dose coconut water vinegar)	Induced obese mice given HFD + 0.08 ml/kg body weight coconut water vinegar
CH (high-dose coconut water vinegar)	Induced obese mice given HFD + 2 ml/kg body weight coconut water vinegar

Table 2. Primer sequences used in the quantitative real-time polymerase chain reaction assay.

Target genes
AD	Forward: 5ʹ-TCAGGAAGAGGAGGAGGA-3ʹ
	Reverse: 5ʹ-TCAGGAAGCACATCATACG-3ʹ
SREBP1c	Forward: 5ʹ-TCATCAACAACCAAGACAGT-3ʹ
	Reverse: 5ʹ-CCAGAGAAGCAGAAGAGAAG-3ʹ
GLUT4	Forward: 5ʹ-CTGCTTCTGGCTCTCACA-3ʹ
	Reverse: 5ʹ-AGGACATTGGACGCTCTC-3ʹ
iNOS	Forward: 5ʹ-GCACCGAGATTGGAGTTC-3ʹ
	Reverse: 5ʹ-GAGCACAGCCACATTGAT-3ʹ
NF-κB	Forward: 5ʹ-CATTCTGACCTTGCCTATCT-3ʹ
	Reverse: 5ʹ-CTGCTGTTCTGTCCATTCT-3ʹ
Reference genes
GAPDH	Forward: 5ʹ-TTCCAGCCTTCCTTCTTG-3ʹ
	Reverse: 5ʹ-GGAGCCAGAGCAGTAATC-3ʹ
ACTB	Forward: 5ʹ-GAAGGTGGTGAAGCAGGCATC-3ʹ
	Reverse: 5ʹ-GAAGGTGGAAGAGTGGGAGTT-3ʹ
HPRT	Forward: 5ʹ-CGTGATTAGCGATGATGAAC-3ʹ
	Reverse: 5ʹ-AATGTAATCCAGCAGGTCAG-3ʹ

AD, adiponectin; SREBP1c, sterol regulatory element-binding protein-1c; GLUT4, glucose transporter type 4; iNOS, inducible nitric oxide synthase; NF-κB, nuclear factor-κB; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ACTB, beta-actin; HPRT, hypoxanthine phosphoribosyltransferase.

Table 3. Body weight, fat-pad weight, and fat-pad/body weight ratio of untreated (UT), 0.08 ml/kg body weight (BW) coconut water vinegar (CL) and 2 ml/kg BW coconut water vinegar (CH)-treated high-fat-diet (HFD)-induced obese mice at the end of week 33.

Group	Final BW (g/mouse)	Fat-pad weight	Fat-pad/BW ratio (%)
UT	53.30 ± 0.50^a	2.43 ± 0.14^a	4.56 ± 0.07^a
CL	48.64 ± 1.74^a	2.11 ± 0.04^a	4.82 ± 0.05^a
CH	43.76 ± 1.46 ^b	1.31 ± 0.25^b	2.54 ± 0.49^b

Data are shown as mean ± SD of biologically replicated mice from the same treatment group.

Different superscript letters indicate significant differences among the groups (p < 0.05).

Figure 1. Body weight measurement (week 0–33) of untreated mice and mice fed 0.08 ml/kg body weight (BW) coconut water vinegar (CL) or 2 ml/kg BW coconut water vinegar (CH). Data are shown as the average of biological replicates of mice from the same treatment group. Different letters indicate significant differences among groups (p < 0.05).

Table 4. Serum biochemical profile of untreated (UT), 0.08 ml/kg body weight (BW) coconut water vinegar (CL) and 2 ml/kg BW coconut water vinegar (CH)-treated high-fat-diet (HFD)-induced obese mice.

Group	Cholesterol (mmol/l)	Triglyceride (mmol/l)	LDL (mmol/l)	HDL (mmol/l)	HDL/LDL
UT	5.21 ± 0.15^a	6.27 ± 0.71^a	1.26 ± 0.12^a	1.10 ± 0.15^a	0.87 ± 0.13^a
CL	3.71 ± 0.46^b	2.95 ± 0.55^b	0.95 ± 0.11^a	1.72 ± 0.12^a	1.81 ± 0.09^a
CH	3.67 ± 0.21^b	2.39 ± 0.22^c	0.75 ± 0.02^b	3.03 ± 0.11^b	3.20 ± 0.15^b

Data are shown as mean ± SD of biologically replicated mice from the same treatment group.

LDL, low-density lipoprotein; HDL, high-density lipoprotein.

Different superscript letters indicate significant differences among the groups (p < 0.05).

Figure 2. (a) Quantitative polymerase chain reaction analysis of obesity-related genes [glucose transporter type 4 (Glut4), adiponectin, and sterol regulatory element-binding protein-1 (SREBP1)] of untreated (UT) mice and mice fed 0.08 ml/kg body weight (BW) coconut water vinegar (CL) or 2 ml/kg BW coconut water vinegar (CH). (b) Significant adipokines down-regulated by coconut water vinegar treatment in high-fat-diet obese mice tested using adipokine proteome profiler analysis. RBP4, retinol-binding protein-4. Data are presented as the mean ± SD of biological replicates of mice from the same treatment group. Different letters indicate significant differences among groups (p < 0.05).

Figure 3. (a) Nitric oxide (NO) content and (b) quantitative polymerase chain reaction analyses of inflammation related genes [nuclear factor-κB (NFκB) and inducible nitric oxide synthase (iNOS)] of untreated (UT) mice and mice fed 0.08 ml/kg body weight (BW) coconut water vinegar (CL) or 2 ml/kg BW coconut water vinegar (CH). Data are presented as the mean ± SD of biologically replicated mice from the same treatment group. Different letters indicate significant differences among groups (p < 0.05).

Figure 4. Comparison of the relative abundance of gut microbiota at genus level of untreated (UT) mice and mice fed 2 ml/kg body weight coconut water vinegar (CH) at the end of week 33 post-experiment using Illumina 16S rRNA metagenomic sequencing. Data are presented as the mean ± SD of biologically replicated mice (n = 6) from the same treatment group. Different letters indicate significant differences between groups (p < 0.05).