A metabolic mechanism analysis of Fuzheng-Huayu formula for improving liver cirrhosis with traditional Chinese medicine syndromes

Abstract

Fuzheng-Huayu formula (FZHY), a Chinese herbal mixture prescription, has been proven effective in treating liver fibrosis and cirrhosis in both clinical trials and animal experiments. In this study we assessed the metabolic mechanisms of traditional Chinese medicine (TCM) syndrome-based FZHY treatment in liver cirrhosis (LC). A total of 113 participants, including 50 healthy controls and 63 LC patients, were recruited. According to the diagnosis and differentiation of the TCM syndromes, the LC patients were classified into 5 TCM syndrome groups including the liver stagnation syndrome (LSS), spleen deficiency and damp overabundance syndrome (SDDOS), damp-heat accumulation syndrome (DHAS), liver-kidney Yin deficiency syndrome (LKYDS), and blood stagnation syndrome (BSS), and administered FZHY for 6 months. FZHY treatment significantly decreased serum levels of hyaluronic acid (HA), a biochemical marker for LC, as well as TCM syndrome scores (the TCM syndrome scores were decreased in all the groups with significant decreases in the LSS and LKYDS groups). Furthermore, FZHY treatment gradually shifted the metabolic profiles of LC patients from a pathologic state to a healthy state, especially in LC patients with LSS and LKYDS. Twenty-two differently altered metabolites (DAMs) were identified, including carbohydrates, amino acids, fatty acids, etc with 9 DAMs in LSS patients, 9 in LKYDS patients, and 4 in other patients. The metabolic pathways involved in the conversion of amino acids and the body's detoxification process were regulated first, followed by the pathways involved in the body's energy supply process. In conclusion, the evaluation of the effect of TCM syndrome-based FZHY treatment show that FZHY has a better effect on LKYDS and LSS than on the other TCM syndromes, and the metabolic mechanisms might be involved in the increased detoxification function in LKYDS and the improvement of energy supply in LSS, which provides important evidence for the clinical application of TCM syndrome-based treatment.

Figure 1

Clinical parameters of healthy controls, LC patients without and with six month-treatment of FZHY. n=50 in healthy group, and n=63 in LC group. Values were expressed as mean±SD. Significant differences were analyzed by One-Way ANOVA test or Student's t test. Compared with healthy group, ^*P<0.05, ^**P<0.01 compared with LC group. ^##P<0.01.

Figure 2

TCM syndrome scores of LSS group, SDDOS group, DHAS group, LKYDS group and BSS group before and after FZHY treatment. TCM syndrome scores were calculated according to 45 clinical symptoms and 39 signs. n=17 in LSS group, n=10 in SDDOS group, n=11 in DHAS group, n=14 in LKYDS group and n=11 in BSS group. Compared with LC group. ^**P<0.01.

Figure 3

Dynamic changes of metabolomic profiling in healthy group, LC and FZHY groups with different time of FZHY treatment. PLS-DA analysis was performed and score plot was shown. Black referred to healthy group, red refers to LC group (LC-0M), blue referred to FZHY group with one month of FZHY treatment (LC-1M), green referred to FZHY group with three months of FZHY treatment (LC-3M), and orange referred to FZHY group with six months of FZHY treatment (LC-6M).

Figure 4

Dynamic changes of metabolomic profiling in LSS, LKYDS and the other TCM syndromes (others) by FZHY treatment. PLS-DA analysis was performed and score plots were shown. Black referred to healthy group, red referred to LC group (LC-0M), blue referred to FZHY group with one month of FZHY treatment (LC-1M), green referred to FZHY group with three months of FZHY treatment (LC-3M), and orange referred to FZHY group with six months of FZHY treatment (LC-6M).

Figure 5

Dynamic changes of significantly altered metabolites among healthy group, LC group and FZHY groups, involved in carbohydrate, amino acid, and lipid metabolism. n=50 in healthy group and n=63 in LC group. Values were expressed as mean±standard deviation. Significant differences were analyzed by One-Way ANOVA test. Compared with healthy group, ^**P<0.01; compared with LC group. ^#P<0.05, ^## P<0.01 vs FZHY-1M and FZHY-3M.

Figure 6

DAMs in different TCM syndromes. Nine DAMs were found in LSS group, nine DAMs were found in LKYDS group, and only four were found in the other TCM syndromes group.

Figure 7

The network of significantly altered metabolic pathways in response to FZHY treatment. Red metabolites referred to ones significantly altered after LC patients took one month of FZHY, yellow metabolites referred to ones significantly altered after LC patients took three months of FZHY, and green metabolites refer to ones significantly altered after LC patients took six months of FZHY. Metabolites which changed in the first month mostly belonged to amino acid metabolism, located in red box. Metabolites which changed in the third or sixth month mostly belonged to carbohydrate and lipid metabolism, located in green box.

Figure 8

The network of significantly altered metabolic pathways in LSS and LKYDS. The significantly altered metabolic pathways in red box belong to LSS, which were mainly involved in carbohydrate metabolism and energy supply. The pathways in blue box belong to LKYDS, which were mainly involved in amino acid metabolism and detoxification. Red metabolites referred to ones significantly up-regulated by FZHY treatment, and blue metabolites referred to ones significantly down-regulated by FZHY treatment.