Dietary herbaceous mixture supplementation reduced hepatic lipid deposition and improved hepatic health status in post-peak laying hens

Abstract

Fatty liver hemorrhagic syndrome is characterized by hepatic damage and hemorrhage impairing animal welfare in birds, which was well-known to be moderately relieved through dietary choline chloride supplementation in laying hens. Chinese herb has been proven to exert a positive role on hepatic health in human and rodents. Here, we investigated the effect of herbaceous mixture (HM), which consists of Andrographis paniculate, Silybum marianum, Azadirachta Indica, and Ocimum basilicum (2:3.5:1:2), on the hepatic lipid metabolism and health status in laying hens. A total of 240 Hy-line Brown hens (389-day-old) were randomly fed the basal diet with 0 mg/kg choline chloride (negative control, NC), 1,000 mg/kg choline chloride (control, Ctrl), or 300 mg/kg HM for 28 d. Birds fed HM diet exhibited lower serum triglyceride (TG) and low-density lipoprotein cholesterol concentration, and higher high-density lipoprotein cholesterol level than those received NC and Ctrl diets (P < 0.05). When compared to control and NC group, the diets with HM decreased the contents of total cholesterol and TG in liver, as well as upregulated the mRNA abundance of hepatic hormone-sensitive lipase and lipoprotein lipase. Meanwhile, the hepatic area and diameter of steatosis vacuoles were also decreased by dietary HM administration (P < 0.05), which accompanied by decreased serum alanine aminotransferase activity (P < 0.05). Birds fed HM diets enhanced the hepatic antioxidative capacity than those received NC and Ctrl diet. Dietary HM depressed the mRNA level of inflammatory cytokine as compared to NC but not Ctrl group. Collectively, the diet with 300 mg/kg HM has a favorable effect in decreasing the lipid deposition and protecting liver injury by alleviating hepatic oxidant stress and inflammation in post-peak laying hens.

Keywords: herbaceous mixture; lipid deposition; liver injury; post-peak laying hen.

Figure 1

The effect of dietary herbaceous mixture (HM) supplementation on the production performance of laying hens including (A) egg production, (B) egg weight, (C) average daily feed intake, and feed conversion ration expressed as the ratio of grams of total egg weight to grams of total feed intake. Values are presented as mean ± standard error (n = 6). Different letters in the same column represent the significant difference (P < 0.05). Abbreviations: Ctrl, choline chloride control group; NC, negative control.

Figure 2

Herbaceous mixture (HM) improved serum lipid profile in post-peak laying hens. The concentration of (A) triglyceride, (B) total cholesterol, (C) high density lipoprotein cholesterol (HDL-c), and (D) low density lipoprotein cholesterol (LDL-c) in serum. Values are presented as mean ± standard error (n = 6). Different letters in the same column represent the significant difference (P < 0.05). Abbreviations: Ctrl, choline chloride control group; NC, negative control.

Figure 3

Hepatic lipid metabolism response to dietary herbaceous mixture (HM) treatment in post-peak laying hens. (A) Liver oil red O staining (scale: 50 μm), the concentration of (B) triglyceride, (C) total cholesterol, and (D) free fatty acid (FFA) of liver (n = 8). mRNA abundance of (E) lipogenesis including fatty acid synthase (FAS), acetyl CoA carboxylase 1 (ACC-1), sterol regulatory element binding transcription factor 1 (SREBP-1), and stearoyl-CoA desaturase (SCD), (F) lipid transportation, i.e., microsomal triglyceride transfer protein (MTTP), low density lipoprotein receptor (LDLR), and very low-density lipoprotein receptor (VLDLR), and (G) lipolysis including hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) (n = 6). Values are presented as mean ± standard error. Different letters in the same column represent the significant difference (P < 0.05). Abbreviations: Ctrl, choline chloride control group; NC, negative control.

Figure 4

Herbaceous mixture (HM) attenuated liver steatosis and hepatocyte apoptosis in post-peak laying hens. (A) Liver hematoxylin and eosin (H&E) staining (scale: 50 μm) was performed, and (B) frequency distribution of vacuole, (C) the diameter and (D) area were quantified based on the H&E section (n = 8). The mRNA expression of genes involved in apoptosis including (E, F) Beclin-1, B-cell leukemia/ lymphoma 2 (Bcl-2), Caspase 3, and Caspase 8 were measured by RT-PCR (n = 6). (G) Serum aspartate transaminase (AST) was determined (n = 7). Values are presented as mean ± standard error. Different letters in the figure represent the significant difference (P < 0.05). Abbreviations: Ctrl, choline chloride control group; NC, negative control.

Figure 5

Effects of herbaceous mixture (HM) on hepatic oxidative stress and inflammation in post-peak laying hens. The antioxidative stress was evaluated through (A) malondialdehyde (MDA), (B) total antioxidative capacity (T-AOC), (C) superoxide dismutase (SOD), (D) glutathione peroxidase (GSH-Px), and (E) catalase (CAT); RT-PCR analysis of proinflammatory cytokines including (F) tumor necrosis factor-α (TNF-α) and (G) interleukin-6. Values are presented as mean ± standard error (n = 6). Different letters in the same column represent the significant difference (P < 0.05). Abbreviations: Ctrl, choline chloride control group; NC, negative control.