Nitrosamine exposure exacerbates high fat diet-mediated type 2 diabetes mellitus, non-alcoholic steatohepatitis, and neurodegeneration with cognitive impairment

Table 2 Assays of Liver Injury or Hepatic Steatosis

Assay^§	LFD+VEH	LFD+NDEA	HFD+VEH	HFD+NDEA	F-Ratio	P-Value
Serum ALT (U/L)	17.3 ± 0.9	23.8 ± 3.9	30.6 ± 1.8**	33.5 ± 2.1**	10.08	< 0.0001
Nile Red (RFU/μg)	111.0 ± 2.6	128.3 ± 15.0	122.7 ± 6.1	147.9 ± 6.1*	2.77	0.06
Cholesterol (ng/mg)	360.5 ± 20.2	340.1 ± 33.9	435.4 ± 21.9	458.4 ± 19.7*	5.02	0.011
Triglycerides (μg/mg)	11.2 ± 1.5	10.8 ± 2.0	17.1 ± 0.8*	21.0 ± 1.4**	10.3	0.0003

§ALT = alanine transaminase, Nile Red, cholesterol, and tryglyceride levels were measured in liver tissue. RFU/μg = relative fluorescence units; Nile red, cholesterol and triglyceride measurements are relative to liver. Inter-group comparisons were made using ANOVA tests. For all measurements, df = 3/36. Significant differences relative to the LFD control group were demonstrated using the Dunnett's post-hoc multiple comparison test (*P < 0.05; **P < 0.01; ***P < 0.001).

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