Effect of insulin-sensitizing agents in combination with ezetimibe, and valsartan in rats with non-alcoholic fatty liver disease

Table 1 Liver-related parameters, plasma biochemistry and cytokines in rats fed with methionine choline-deficient diet (MCDD) or choline-supplemented diet (Chow diet) (mean ± SD, n = 6)

Parameters	CL	FL	P values
	Chow diet	MCDD diet
Baseline rat mass (g )	227 ± 7	226 ± 6	0.5
Final rat mass (g )	406 ± 19	257 ± 12	0.03
Final liver mass (g )	10.2 ± 0.3	10.8 ± 0.8	0.4
Liver/body mass ratio	0.025 ± 0.003	0.042 ± 0.003	0.02
% change		68
Liver triglycerides (mol/g)	1.6 ± 0.3	21.8 ± 1.0	< 0.001
% change		1263
Liver cholesterol (mol/g)	0.5 ± 0.0	1.6 ± 0.2	< 0.03
% change		245
Serum HOMA-IRI	7 ± 0.3	18 ± 2.0	< 0.001
Liver tocopherol/MDA ratio	0.05	0.003	< 0.03
FL infiltration (%)	0	91 ± 5	< 0.001
ALT (nkat/L)	667 ± 83	800 ± 33	< 0.05

CL: Control liver; FL: Fatty liver; HOMA-IRI: Insulin resistance index; MDA: Malondialdehyde.

Table 2 Plasma cytokines, lipids, and biochemical parameters at the end of the study period (wk 15)

Parameters	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	P
	FL	Rosiglitazone	Metformin	M+R	Ezetimibe	Valsartan	R+M+V	R+M+V+E
% changes W/BW	+68	+12	+28	+20	+28	+16	+20	+28	< 0.001
ALT ( nkat/L)	800 ± 33	684 ± 16.5	650 ± 33	667 ± 16.5	684 ± 49.5	734 ± 49.5	684 ± 33	717 ± 33	NS
ALB (g/L)	33 ± 7	34 ± 8	33 ± 16	34 ± 15	33 ± 6	33 ± 8	38 ± 5	35 ± 2	NS
Cholesterol (mmol/L)	123 ± 13	148 ± 13	145 ± 17	106 ± 15	100 ± 7	121 ± 16	139 ± 10	97 ± 17	< 0.001
Triglyceride (mmol/L)	0.8 ± 0.1	0.33 ± 0.08	0.5 ± 0.1	0.4 ± 0.1	0.35 ± 0.07	0.4 ± 0.04	0.5 ± 0.06	0.43 ± 0.8	< 0.03
TNF-α (ng/mL)	17.8 ± 10.0	19 ± 15	10 ± 6	30 ± 26	12 ± 7	9 ± 7	10 ± 6	13 ± 12	< 0.01
HOMA-IR	18 ± 2	11 ± 1	12 ± 1	9 ± 0.5	13 ± 1	16 ± 2	9 ± 0.5	8 ± 1	< 0.001
% change	+157	-39	-33	-50	-28	-11	-50	-55

Table 3 Hepatic lipid composition, MDA, and alpha-tocopherol levels (mean ± SD, n = 6)

Parameters	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	P
	FL	Rosiglitazone	Metformin	M+R	Ezetimibe	Valsartan	R+M+V	R+M+V+E
Triglyceride (mol/g liver)	21.8 ± 1.0	9.9 ± 1.4	13.1 ± 1.4	7.9 ± 1.9	10.1 ± 1.5	9.9 ± 1.7	6.9 ± 1.2	7.9 ± 2.1	< 0.0001
% change	+1263	-54	-40	-64	-53	-54	-68	-64
Cholesterol (mol/g liver)	1.6 ± 0.2	1.6 ± 0.2	1.6 ± 0.1	1.2 ± 0.2	1.2 ± 0.4	1.9 ± 0.3	1.3 ± 0.3	1.1 ± 0.4
% change	+245	0	0	-27	-25	+19	-19	-31	< 0.003
MDA (mol/g liver)	2.01 ± 0.04	1.2 ± 0.1	0.7 ± 0.1	0.9 ± 0.01	1.1 ± 0.1	1.2 ± 0.2	0.8 ± 0.1	0.6 ± 0.1	< 0.001
% change	+235	-40	-65	-55	-45	-40	-60	-70
Tocopherol- α (mg/g liver)	0.07 ± 0.03	0.13 ± 0.04	0.2 ± 0.1	0.2 ± 0.04	0.34 ± 0.09	0.14 ± 0.02	0.19 ± 0.04	0.38 ± 0.8	< 0.001
% change	-77	+86	+186	+186	+386	+100	+171	+443
Tocopherol-α /MDA ratio	0.003	0.01	0.028	0.02	0.03	0.011	0.023	0.06	< 0.001
% change	-94	+233	+833	+566	+900	+266	+666	+1900

Table 4 Liver histological injury: Grading and staging (mean ± SD, n = 6)

Liver histology	Chow	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	MCDD	P
		Only	Rosiglitzone	Metformin	M+R	Ezetimibe	Valsartan	R+M+V	R+M+V+E
Steatosis %	0	91 ± 5	45 ± 5	74 ± 11	59 ± 11	77 ± 8	63% ± 8	51 ± 4	37 ± 15	< 0.001
Lobular inflammation	0.5 ± 0.1	0.7 ± 0.1	0.7 ± 0.1	0.6 ± 0.1	0.6 ± 0.1	0.7 ± 0.1	0.5 ± 0.1	0.5 ± 0.1	0.6	NS
Portal inflammation	None	None	None	None	None	None	None	None	None	NS
Fibrosis (> F1)	None	None	None	None	None	None	None	None	None	NS
# Rats w/MB	0	2	1	2	2	1	1	1	1	NS
Ballooning	0	2	1	1	1	0	1	1	2	NS
Pericellular fibrosis	0	1	0	1	2	1	0	2	1	NS

MB: Mallory body; NS: Non-significant.