Herbal adaptogens combined with protein fractions from bovine colostrum and hen egg yolk reduce liver TNF-α expression and protein carbonylation in Western diet feeding in rats

Abstract

BackgroundWe examined if a purported anti-inflammatory supplement (AF) abrogated Western-diet (WD)-induced liver pathology in rats. AF contained: 1) protein concentrates from bovine colostrum and avian egg yolk; 2) herbal adaptogens and antioxidants; and 3) acetyl-L-carnitine.MethodsNine month-old male Brown Norway rats were allowed ad libitum access to WD for 41–43 days and randomly assigned to WD + AF feeding twice daily for the last 31–33 days (n = 8), or WD and water-placebo feeding twice daily for the last 31–33 days (n = 8). Rats fed a low-fat/low-sucrose diet (CTL, n = 6) for 41–43 days and administered a water-placebo twice daily for the last 31–33 days were also studied. Twenty-four hours following the last gavage-feed, liver samples were analyzed for: a) select mRNAs (via RT-PCR) as well as genome-wide mRNA expression patterns (via RNA-seq); b) lipid deposition; and, c) protein carbonyl and total antioxidant capacity (TAC). Serum was also examined for TAC, 8-isoprostane and clinical chemistry markers.ResultsWD + AF rats experienced a reduction in liver Tnf-α mRNA (-2.8-fold, p < 0.01). Serum and liver TAC was lower in WD + AF versus WD and CTL rats (p < 0.05), likely due to exogenous antioxidant ingredients provided through AF as evidenced by a tendency for mitochondrial SOD2 mRNA to increase in WD + AF versus CTL rats (p = 0.07). Liver fat deposition nor liver protein carbonyl content differed between WD + AF versus WD rats, although liver protein carbonyls tended to be lower in WD + AF versus CTL rats (p = 0.08). RNA-seq revealed that 19 liver mRNAs differed between WD + AF versus WD when both groups were compared with CTL rats (+/- 1.5-fold, p < 0.01). Bioinformatics suggest that AF prevented WD-induced alterations in select genes related to the transport and metabolism of carbohydrates in favor of select genes related to lipid transport and metabolism. Finally, serum clinical safety markers and liver pathology (via lesion counting) suggests that chronic consumption of AF was well tolerated.ConclusionsAF supplementation elicits select metabolic, anti-inflammatory, and anti-oxidant properties which was in spite of WD feeding and persisted up to 24 hours after receiving a final dose.

Western diet-induced hepatic steatosis and alterations in the liver transcriptome in adult Brown-Norway rats

“…Phenotypic changes in WD versus CTL rats are presented in Table 1 below. As reported previously [ 10 ], WD feeding significantly increased body mass at sacrifice by 6 % ( p = 0.0019), omental fat pad mass by 52 % ( p < 0.001), perirenal fat pad mass by 74 % ( p < 0.001), and liver fat deposition by 4.2-fold ( p = 0.004). We also previously reported that WD rats consumed 15 % more calories compared to CTL rats ( p < 0.001), with a diet higher in total fat, total sucrose, and total cholesterol.…”

“… Values are means ± SE. Body mass, total kcal consumed, and Oil-Red O data previously reported [ 10 ]; symbols: g, grams; AUs, arbitrary units …”

“…Food intakes were carefully measured weekly (i.e., monitoring cage bottoms for morsels, etc.) and total caloric intakes for the duration of the study have been reported elsewhere [ 10 ].…”

“…Euthanasia procedures have been described in detail elsewhere [ 10 ]. Briefly, on the day of euthanasia, animal cages were removed from the animal quarters between 0800–0900 and food was removed from each cage.…”

“…We recently performed a sub-chronic (6-week) feeding study examining the effects of an antioxidant supplement on markers of liver health during concomitant WD feeding in 9-month old adult Brown-Norway rats [ 10 ]. When comparing WD ad libitum -fed rats versus rats provided control diet (CTL; low fat, low sugar) groups, we found pronounced hepatic lipid accumulation, as determined by Oil Red O and H&E staining, suggestive of a NAFLD phenotype.…”