Effect of Fuzheng Huayu formula and its actions against liver fibrosis

Abstract

Liver fibrosis is a common histological process to develop into cirrhosis in various chronic liver diseases including chronic hepatitis and fatty liver. Therefore anti-liver fibrosis is very important strategy to treat chronic liver diseases. Fuzheng Huayu (FZHY), a preparation containing herbs such as Radix Salvia Miltiorrhizae, Cordyceps, Semen Persicae, was formulated on the basis of Chinese medicine theory in treating liver fibrosis and was approved. Pharmacological studies and clinical trials demonstrate that FZHY has a significant effect against liver fibrosis and that many of the pharmacological actions are attributable to the effect. This article reviews the effects and actions of FZHY, in particular the effects observed from clinical trials in treating liver fibrosis caused by chronic hepatitis B and the actions on inhibition of hepatic stellate cell activation, protection of hepatocytes and inhibition of hepatic sinusoidal capillarization. This article also reviews the coordinated effects of the constituent herbs of FZHY and the actions of their active compounds such as salvianonic acid B (SA-B) on liver fibrosis.

Figure 1

Effect of FZHY on liver fibrosis in a patient with chronic hepatitis B. The biopsy liver tissues were stained with VG before (A) and after (B) treatment. × 100.

Figure 2

Effect of FZHY on α-SMA expression in fibrotic liver. The liver fibrosis was induced with CCl₄ in rats. Treatment group was administered with FZHY at the start of six-week intoxication, while the control group was given saline; α-SMA expression in liver tissue was determined by immunohistochemical stain. (×100).

Figure 3

Effect of FZHY drug serum on collagen secretion from HSC. The collagen secretion rate was assayed by [³H]-Proline incorporation and collagenase digestion; values are expressed as mean ± standard deviation (SD) of three separate experiments and compared by t test. *P < 0.05 vs. the control. 5%, 10% and 20% mean different concentrations of sera from rats administered with FZHY (FZHY drug serum) or saline (control).

Figure 4

Effect of FZHY on TGF-β1 expression in activated HSC. Immunocytochemical stain with anti-TGF-β1 (×200). A: HSC treated with normal rat serum (control); B: HSC treated with FZHY drug serum.

Figure 5

Effect of FZHY drug serum treated Kuppfer cell conditioned medium on quiescent HSC proliferation and Col-I secretion. HSC proliferation was assayed with [³H]TdR incorporation HSC; type I collagen secretion was measured at HSC supernatants with ELISA. Values are expressed as mean ± standard deviation (SD) of three separate experiments and compared by ANOVA. *P < 0.05, vs. N-KcCM, #P < 0.05, vs. C-KcCM.

Figure 6

Effect of FZHY on FN and integrin pathway mediators. Liver fibrosis was induced by DMN and treated with FZHY. Normal: normal rats; Model, model control; FZHY: FZHY-treated. Protein expression and FAK phosphorylation were checked with Western blot.

Figure 7

Dynamic changes of MMP2/9 activity and effect of FZHY on fibrotic liver of CCl₄ induced rat. Gelatin zymography. Time phases after CCl₄ intoxication. N: normal; M: model control; FZHY: FZHY-treated.

Figure 8

Effect of FZHY on hepatocytes apoptosis in vivo. Acute liver injury with hepatocyte apoptosis was induced by infusion of LPS plus D-GalN for six hours in mice. Treatment group was administered with FZHY two days before LPS intoxication, while the control was given saline. The apoptotic hepatocytes were stained with TUNEL. (×100).

Figure 9

Effect of FZHY drug serum on apoptosis in hepatocytes in vitro. Primary hepatocytes were isolated from mice and cultured; apoptosis was induced by TNF-α and actinomycin D (Act D) for six hours. Control group was incubated with normal rat serum, while treatment group was administered with FZHY drug serum at the same time. Normal control was cultured with newborn calf serum without TNF-α and Act D. The cell apoptosis was checked with Annexin V/Propidium Iodide (PI) stain and observed under the confocal laser scanning microscopy. Early apoptotic cells were Annexin V positive (green) alone, late apoptotic and necrotic cells were both Annexin V and PI positive or PI positive (red) alone respectively. (×630).

Figure 10

Effect of SA-B histological changes of liver fibrosis in chronic hepatitis B. The liver biopsy examination before treatment (A: S4) and after treatment (B: S3), stained with Gorden-Sweet and Masson trichrome method. (×100).