Effects of combined dietary supplementation with fenofibrate and Schisandrae Fructus pulp on lipid and glucose levels and liver function in normal and hypercholesterolemic mice

Figures & data

Figure 1 Experimental design of the study.

Abbreviations: SF-P, Schisandrae Fructus pulp; AqSF-P, aqueous extract of SF-P; BY, bicyclol; TC, total cholesterol; TG, triglyceride; HDL, high-density lipoprotein; LDL, low-density lipoprotein; N-HDL, non-high-density lipoprotein; ALT, alanine aminotransferase; ND, normal diet; HCBD, high cholesterol/bile salt diet.

Table 1 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on serum lipid profiles in ND- and HCBD-fed mice

Groups	Dose (%/w/w)	TC (mmol/L)	TG (mmol/L)	HDL (mmol/L)	LDL (mmol/L)	HDL/LDL	LDL/HDL	N-HDL (mmol/L)
ND-fed mice
ND	n/a	4.31±0.21	2.04±0.18	4.19±0.21	0.31±0.01	13.68±5.37	0.07±0.00	0.21±0.09
FF	0.1	3.38±0.23**	0.93±0.07**	3.27±0.19**	0.31±0.02	10.54±0.37**	0.10±0.00**	0.31±0.06
FF/BY	0.1/0.025	2.76±0.28**	0.78±0.11**	2.58±0.14**,†	0.28±0.02	9.40±0.51**	0.11±0.01**	0.41±0.27
FF/AqSF-P	0.1/0.3	3.17±0.17**	0.88±0.08**	3.06±0.16**	0.30±0.02	10.42±0.28**	0.10±0.00**	0.23±0.07
	0.1/1	3.44±0.14**	0.95±0.09**	3.05±0.14**	0.28±0.01	11.06±0.32*	0.09±0.00**	0.39±0.04**
	0.1/3	3.24±0.26**	0.81±0.08**	2.93±0.23**	0.30±0.02	9.90±0.56**	0.10±0.01**	0.32±0.07
	0.1/9	3.35±0.26*	0.73±0.09**	3.19±0.24**	0.30±0.03	10.75±0.60**	0.10±0.01**	0.60±0.25
HCBD-fed mice
ND	n/a	4.30±0.27	2.33±0.11	4.25±0.22	0.52±0.05	8.58±0.47	0.12±0.01	0.32±0.08
HCBD	n/a	6.38±0.25**	1.24±0.11**	4.37±0.14	2.55±0.09**	1.72±0.05**	0.59±0.02**	2.00±0.20**
FF	0.03	3.94±0.25##	0.98±0.08	3.43±0.23##	1.07±0.11##	3.33±0.17##	0.31±0.02##	0.51±0.11##
FF/BY	0.03/0.025	4.61±0.18#,†	0.93±0.05#	4.02±0.15†	1.13±0.09##	3.71±0.23##	0.28±0.02##	0.59±0.11##
FF/AqSF-P	0.03/3	4.28±0.20##	0.86±0.06##	3.74±0.22#	0.99±0.06##	3.83±0.24##	0.27±0.02##	0.53±0.08##
	0.03/9	3.87±0.13##	0.85±0.05##	3.45±0.13##	0.90±0.06##	3.97±0.25##	0.26±0.01##,†	0.58±0.05##
HCBD-fed mice
ND	n/a	4.29±0.12	2.18±0.10	3.80±0.10	0.46±0.04	8.71±0.53	0.12±0.01	0.55±0.04
HCBD	n/a	5.28±0.28**	1.22±0.08**	3.60±0.18	1.35±0.16**	3.51±0.96**	0.38±0.04**	1.68±0.14**
FF	0.1	3.99±0.19##	1.12±0.06	3.16±0.16	0.82±0.10#	4.25±0.48	0.27±0.04	0.99±0.12##
FF/BY	0.1/0.025	3.84±0.25##	1.04±0.08	3.04±0.21	0.63±0.08##	5.21±0.45	0.21±0.02##	0.80±0.05##
FF/AqSF-P	0.1/1	3.89±0.27##	1.08±0.08	3.07±0.24	0.57±0.07##	5.61±0.30†	0.18±0.01##,†	0.83±0.14##
	0.1/3	3.95±0.15##	1.14±0.08	2.96±0.11##	0.87±0.09#	3.71±0.39	0.30±0.03	0.99±0.10##
	0.1/9	4.18±0.14##	1.06±0.08	3.08±0.18	0.89±0.10#	3.99±0.70	0.29±0.03	1.10±0.13##

Notes: Mice were fed with ND or HCBD to establish a model of hypercholesterolemia. FF or the combination of FF and AqSF-P or BY at the indicated doses were supplemented with either ND or HCBD. The concentrations of AqFS-P were estimated on the basis of crude herbal material. Ten days after dietary supplementation, serum TC, TG, HDL, LDL, and N-HDL levels, as well as HDL/LDL and LDL/HDL ratios, were measured. Values given are the means ± SEM, with n=10.

* P<0.05

** P<0.01 vs ND

# P<0.05

## P<0.01 vs HCBD

† P<0.05 vs FF alone. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.

Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; w/w, weight/weight; TC, total cholesterol; TG, triglyceride; HDL, high-density lipoprotein; LDL, low-density lipoprotein; N-HDL, non-high-density lipoprotein; SEM, standard error of mean; vs, versus; ANOVA, analysis of variance; n/a, not applicable.

Figure 2 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on hepatic lipid and glucose levels in normal and hypercholesterolemic mice.

Notes: Mice were fed with ND or HCBD without and with supplementation with FF or the combination of FF and AqSF-P or BY at the indicated concentrations. Ten days later, hepatic TC (A), TG (B), and glucose (C) levels were measured. Values given are the means ± SEM, with n=10. *P<0.05, **P<0.01 vs ND; ^##P<0.01 vs HCBD; ^†P<0.05, ^††P<0.01 vs FF alone. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.
Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicylol; TC, total cholesterol; TG, triglyceride; SEM, standard error of mean; ANOVA, analysis of variance; w/w, weight/weight; vs, versus.

Figure 3 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on fecal cholesterol excretion in normal and hypercholesterolemic mice.

Notes: Mice were fed with ND or HCBD without and with supplementation with FF or the combination of FF and AqSF-P or BY at the indicated concentration. Ten days later, fecal TC content was measured. Values given are the means ± SEM, with n=10. *P<0.05, **P<0.01 vs ND; ^#P<0.05, ^##P<0.01 vs HCBD. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.
Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; TC, total cholesterol; TG, triglyceride; SEM, standard error of mean; vs, versus; ANOVA, analysis of variance; w/w, weight/weight.

Table 2 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on hepatic weight, hepatic index, and liver function in ND-and HCBD-fed mice

Groups	Dose (%/w/w)	Hepatic weight (g)	Hepatic index	Serum ALT activity (U/L)
ND-fed mice
ND	n/a	1.71±0.07	6.01±0.16	40.80±7.10
FF	0.1	3.00±0.11**	11.46±0.19**	121.70±14.14**
FF/BY	0.1/0.025	3.18±0.07**	11.81±0.17**	55.60±5.83††
FF/AqSF-P	0.1/0.3	3.19±0.07**	11.87±0.17**	134.60±13.96**
	0.1/1	3.07±0.06**	11.90±0.25**	149.14±23.77**
	0.1/3	3.00±0.14**	11.40±0.28**	127.30±12.15**
	0.1/9	2.97±0.08**	11.67±0.23**	88.30±5.95**
HCBD-fed mice
ND	n/a	1.74±0.13	6.09±0.40	69.40±1.28
HCBD	n/a	2.10±0.07*	7.33±0.11**	81.00±1.05**
FF	0.03	3.51±0.08##	11.61±0.21##	122.80±7.28##
FF/BY	0.03/0.025	3.49±0.12##	11.69±0.21##	71.50±4.51††
FF/AqSF-P	0.03/3	3.25±0.12##	11.09±0.20##	108.70±8.15##
	0.03/9	3.52±0.10##	11.99±0.18##	100.00±4.44##,†
HCBD-fed mice
ND	n/a	1.75±0.04	6.32±0.07	60.00±3.38
HCBD	n/a	2.07±0.03**	7.38±0.09**	86.90±7.48**
FF	0.1	3.69±0.21##	14.48±0.48##	191.13±35.94##
FF/BY	0.1/0.025	3.79±0.12##	14.44±0.24##	150.78±21.79#
FF/AqSF-P	0.1/1	3.77±0.06##	14.31±0.18##	296.71±63.52##
	0.1/3	4.05±0.12##	14.72±0.31##	288.00±38.37##
	0.1/9	3.46±0.12##	14.10±0.32##	378.56±57.42##

Notes: Mice were fed with ND or HCBD to establish a model of hypercholesterolemia. FF or the combination of FF and AqSF-P or BY at the indicated doses were supplemented with either ND or HCBD. The concentrations of AqFS-P were estimated on the basis of crude herbal material. Ten days after the dietary supplementation, hepatic weight/index and serum ALT activity were measured. Hepatic index was calculated from the following equation: hepatic weight/body weight ×100. Values given are the means ± SEM, with n=10.

* P<0.05

** P<0.01 vs ND

# P<0.05

## P<0.01 vs HCBD

† P<0.05

†† P<0.01 vs FF alone. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.

Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; ALT, alanine aminotransferase; SEM, standard error of mean; ANOVA, analysis of variance; vs, versus; w/w, weight/weight; n/a, not applicable.

Table 3 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on the organ indexes in ND- and HCBD-fed mice

Groups	Dose (%/w/w)	Organ weight/body weight ×100			Organ weight/body weight – liver weight ×100
		Fat	Kidney	Testis	Fat	Kidney	Testis
ND-fed mice
ND	n/a	1.23±0.08	1.47±0.04	0.63±0.02	1.32±0.08	1.57±0.05	0.67±0.02
FF	0.1	0.71±0.04**	1.57±0.04	0.68±0.03	0.80±0.05**	1.77±0.04**	0.76±0.04*
FF/BY	0.1/0.025	0.70±0.03**	1.48±0.04	0.65±0.02	0.79±0.03**	1.68±0.05	0.74±0.03
FF/AqSF-P	0.1/0.3	0.69±0.04**	1.59±0.04	0.66±0.03	0.78±0.05**	1.80±0.05**	0.75±0.03*
	0.1/1	0.62±0.05**	1.57±0.03	0.74±0.02**	0.70±0.06**	1.78±0.04**	0.84±0.02**
	0.1/3	0.66±0.05**	1.57±0.04	0.68±0.03	0.75±0.06**	1.77±0.05**	0.77±0.03*
	0.1/9	0.75±0.06**	1.52±0.03	0.76±0.03**	0.85±0.06**	1.72±0.04*	0.86±0.03**
HCBD-fed mice
ND	n/a	1.17±0.07	1.49±0.03	0.47±0.05	1.25±0.08	1.59±0.03	0.50±0.05
HCBD	n/a	0.87±0.05**	1.54±0.03	0.54±0.03	0.94±0.06**	1.66±0.03	0.58±0.03
FF	0.03	0.59±0.03##	1.47±0.03	0.47±0.02	0.67±0.04##	1.67±0.04	0.53±0.02
FF/BY	0.03/0.025	0.54±0.02##	1.51±0.04	0.46±0.02#	0.62±0.02##	1.71±0.04	0.52±0.02
FF/AqSF-P	0.03/3	0.61±0.02##	1.58±0.03†	0.48±0.02	0.69±0.03##	1.78±0.04#,†	0.54±0.02
	0.03/9	0.62±0.03##	1.60±0.05†	0.48±0.01	0.70±0.04##	1.82±0.06#,†	0.54±0.01
HCBD-fed mice
ND	n/a	1.03±0.07	1.63±0.04	0.61±0.02	1.10±0.07	1.75±0.05	0.65±0.02
HCBD	n/a	0.90±0.05	1.62±0.04	0.58±0.02	0.97±0.06	1.75±0.04	0.62±0.03
FF	0.1	0.50±0.04##	1.64±0.07	0.64±0.03	0.58±0.05##	1.92±0.07#	0.75±0.03##
FF/BY	0.1/0.025	0.53±0.02##	1.56±0.05	0.58±0.02	0.62±0.02##	1.82±0.06	0.68±0.03
FF/AqSF-P	0.1/1	0.57±0.03##	1.70±0.04	0.65±0.02	0.67±0.03##	1.98±0.05##	0.75±0.03##
	0.1/3	0.57±0.03##	1.67±0.04	0.58±0.02	0.67±0.04##	1.95±0.05##	0.68±0.03
	0.1/9	0.41±0.03##	1.57±0.04	0.61±0.04	0.48±0.03##	1.83±0.04	0.71±0.05

Notes: Mice were fed with ND or HCBD to establish a model of hypercholesterolemia. FF or the combination of FF and AqSF-P or BY at the indicated doses were supplemented with either ND or HCBD. The concentrations of AqFS-P were estimated on the basis of crude herbal material. Ten days after the dietary supplementation, organ indexes were estimated. Fatty index was obtained from the epididymal fat pad. Values given are the means ± SEM, with n=10.

*P<0.05

**P<0.01 vs ND

# P<0.05

## P<0.01 vs HCBD

† P<0.05 vs FF alone. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.

Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; SEM, standard error of mean; ANOVA, analysis of variance; w/w, weight/weight; vs, versus; n/a, not applicable.

Table 4 Effects of dietary supplementation with FF, FF/AqSF-P, and FF/BY on body weight and food/drug intake in ND- and HCBD-fed mice

Groups	Dose (%/w/w)	Body weight (g) at day 10	Body weight – hepatic weight (g) at day 10	Food intake (g/kg/day)	Drug intake (g/kg/day)
ND-fed mice
ND	n/a	28.45±0.47	26.72±0.41	173.32	n/a
FF	0.1	26.07±0.64**	23.08±0.53**	187.42	0.19
FF/BY	0.1/0.025	26.85±0.38*	23.72±0.31**	189.68	0.19/0.05
FF/AqSF-P	0.1/0.3	26.89±0.56*	23.71±0.51**	199.44	0.20/0.60
	0.1/1	25.87±0.61**	22.81±0.57**	201.74	0.20/2.02
	0.1/3	26.20±0.72*	23.20±0.61**	182.33	0.18/5.47
	0.1/9	25.45±0.54**	22.48±0.48**	193.01	0.19/17.37
HCBD-fed mice
ND	n/a	28.46±0.67	26.72±0.60	195.36	n/a
HCBD	n/a	28.62±0.81	26.53±0.76	184.49	n/a
FF	0.03	30.30±0.72	26.79±0.67	183.83	0.06
FF/BY	0.03/0.025	29.78±0.53	26.29±0.42	178.31	0.05/0.03
FF/AqSF-P	0.03/3	29.24±0.70	26.00±0.59	180.92	0.06/5.47
	0.03/9	29.32±0.52	25.80±0.42	193.72	0.06/17.39
HCBD-fed mice
ND	n/a	27.72±0.49	25.98±0.46	161.84	n/a
HCBD	n/a	28.02±0.24	25.95±0.23	170.91	n/a
FF	0.1	25.42±1.03#	21.72±0.85##	183.91	0.18
FF/BY	0.1/0.025	26.26±0.64#	22.47±0.54##	192.09	0.19/0.05
FF/AqSF-P	0.1/1	26.34±0.43##	22.58±0.39##	179.16	0.18/1.79
	0.1/3	27.51±0.29	23.46±0.20##	195.06	0.19/5.85
	0.1/9	24.50±0.59##	21.04±0.52##	181.28	0.18/16.32

Notes: Mice were fed with ND or HCBD to establish a model of hypercholesterolemia. FF or the combination of FF and AqSF-P or BY at the indicated doses were supplemented with either ND or HCBD. The concentrations of AqFS-P were estimated on the basis of crude herbal material. Ten days after the dietary supplementation, body weight and food/drug intake were estimated. The dosages (g/kg/day) based on crude herbal material were determined with the amount of ingested diet (g/kg/day) and drug concentration in the diet. Values given are the means ± SEM, with n=10.

* P<0.05

** P<0.01 vs ND

# P<0.05

## P<0.01 vs HCBD. Statistically significant differences were determined using a one-way ANOVA, followed by Dunnett’s multiple-comparisons test.

Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; SEM, standard error of mean; ANOVA, analysis of variance; w/w, weight/weight; vs, versus; n/a, not applicable.

Table 5 A summary of the present study

	HCBD (vs ND)	Dietary FF (vs ND or HCBD)		Dietary FF/AqSF-P (vs FF alone)	Dietary FF/BY (vs FF alone)
		0.03%	0.1%
ND-fed mice
Serum TC	n/a	n/a	↓	–	–
Serum TG	n/a	n/a	↓	–	–
Serum HDL	n/a	n/a	↓	–	↓
Serum LDL	n/a	n/a	–	–	–
Serum LDL/HDL			↑	–	–
Serum ALT	n/a	n/a	↑	–	↓
Hepatic TC	n/a	n/a	↓	↓	↓
Hepatic TG	n/a	n/a	↓	↓	–
Hepatic glucose	n/a	n/a	↓	↓	–
Hepatic index	n/a	n/a	↑	–	–
Fecal TC	n/a	n/a	–	–	–
Fat index	n/a	n/a	↓	–	–
Kidney index	n/a	n/a	↑	–	–
Testis index	n/a	n/a	↑	–	–
Body weight gain	n/a	n/a	↓	↑	↑
HCBD-fed mice
Serum TC	↑	↓	↓	–	↑
Serum TG	↓	–	–	–	–
Serum HDL	–	↓	–	–	↑
Serum LDL	↑	↓	↓	–	–
Serum LDL/HDL	↑	↓	–	↓	–
Serum ALT	↑	↑	↑	↓	↓
Hepatic TC	↑	↓	↓	↓	↓
Hepatic TG	↑	↓	↓	↓	↓
Hepatic glucose	↓	↓	↓	↓	↑
Hepatic index	↑	↑	↑	–	–
Fecal TC	↑	–	↑	–	–
Fat index	↓	↓	↓	–	–
Kidney index	–	–	↑	↑	–
Testis index	–	–	↑	–	–
Body weight gain	–	–	↓	↑	–

Notes: ↑, elevated or increased; ↓, inhibited or decreased; –, unaltered.

Abbreviations: ND, normal diet; HCBD, high cholesterol/bile salt (0.5%/0.15%, w/w) diet; FF, fenofibrate; AqSF-P, aqueous extract of Schisandrae Fructus pulp; BY, bicyclol; TC, total cholesterol; TG, triglyceride; HDL, high-density lipoprotein; LDL, low-density lipoprotein; ALT, alanine aminotransferase; vs, versus; n/a, not applicable.