Hepatoprotective effects of a chinese herbal formula, longyin decoction, on carbon-tetrachloride-induced liver injury in chickens

Abstract

The objective of this study is to establish poultry liver injury model induced by (CCl4) and seek effective hepatoprotective herbals for clinical application. Different doses of CCl4 dissolved in vegetable oil (1 : 1, V/V) were injected via pectoral muscle to induce acute liver injury model in chickens. An herbal formula, Longyin decoction, was prepared for hepatoprotection test on chicken acute liver injury models. The pathologic changes of the liver were observed, and the activities of ALT and AST were, respectively, detected to evaluate the hepatoprotective effects of Longyin decoction on chickens. The chicken acute liver injury model was successfully established by injecting CCl4 via pectoral muscle. The best dose of CCl4 inducing chicken liver injury was 4.0 mL/kg·BW (body weight). The results of qualitative determination by HPTLC showed that the components of Longyin decoction contained Gentian, Capillaries, Gardenia, and Bupleurum root. In the high-dose Longyin group and the middle-dose Longyin group, the pathological changes of the damaged liver were mitigated and the activities of ALT and AST in serum were reduced significantly. Longyin decoction has obvious hepatoprotective effect on acute liver injury induced by CCl4.

Figure 1

Chicken liver in group I. The liver is slightly larger and yellow in color.

Figure 2

Chicken liver in group II. The superficial necrosis is found in liver edge.

Figure 3

Chicken liver in group III. The multiple needle tip necrosis appears in liver.

Figure 4

Chicken liver in group IV. The liver is larger and harder and shows large necrotic plaque.

Figure 5

Chicken liver in group V. The liver is larger and harder and shows large necrotic plaque.

Figure 6

Chicken liver in blank control group. The liver is red, shiny, wet, flexible, and keeping normal volume.

Figure 7

Thin-layer chromatogram of Gentian in Longyin decoction. 1: negative control solution free Gnetian; 2: control solution of gentiopicroside; 3: reference drug substance of Gentian; 4–6: sample solutions 1, 2, and 3.

Figure 8

Thin-layer chromatogram of Capillaries in Longyin decoction. 1: negative control solution free Capillaries; 2: reference drug substance of Capillaries; 3–5: sample solution 1, 2, and 3.

Figure 9

Thin-layer chromatogram of Gardenia in Longyin decoction. 1: negative control solution free Gardenia; 2: Control solution of geniposide; 3: reference drug substance of Gardenia; 4–6: sample solution 1, 2, and 3.

Figure 10

Thin-layer chromatogram of Bupleurum root in Longyin decoction. 1: negative control solution free Bupleurum root; 2: control solution of saikosaponin-α; 3: reference drug substance of Bupleurum root; 4–6: sample solution 1, 2, and 3.

Figure 11

HE staining of chicken liver slice in blank control group. The hepatic lobule has complete structure and the liver cell has regular shape, 400x.

Figure 12

HE staining of chicken liver slice in model control group. The hepatocyte tubes arranged haphazardly with severe congestion and inflammatory cell infiltration, liver cells appear necrosis with nuclear fragmentation, 400x.

Figure 13

HE staining of chicken liver slice in high-dose Longyin group. The hepatic lobules have clear structure, the hepatocyte tubes arranged orderly and closely, 400x.

Figure 14

HE staining of chicken liver slice in middle-dose Longyin group. The hepatocyte tubes arranged orderly but slightly loose with mild congestion, the cytoplasm is reduced, 400x.

Figure 15

HE staining of chicken liver slice in low-dose Longyin group. The hepatocyte tubes arranged in disorder with congestion and inflammatory cell infiltration, and the liver cells show pale watery cytoplasm, 400x.

Figure 16

HE staining of chicken liver slice in drug control group. The liver shows congestion and inflammatory cell infiltration, and the liver cell shows granular degeneration, 400x.