Effects of total flavonoids from Nymphaea candida on hepatic fibrosis induced by carbon tetrachloride in rats / 中草药

ABSTRACT

Objective: To investigate the preventive effect of total flavonoids from Nymphaea candida (NCTF) on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods: Wistar rats were randomly divided into control group, model group, NCTF groups (50, 100, 200 mg/kg) and colchicine group (0.2 mg/kg). Rats were subcutaneously injected with 50% CCl4 peanut oil solution (0.1 mL/100g, twice a week for 12 weeks) to induce hepatic fibrosis. In addition to control and model group given with 0.5% CMC-Na, the other groups were intragastrically administered with drugs (1.0 mL/100 g, once a day for 12 weeks), all rats were put to death. Blood, hepatic and splenic tissue were collected to detect liver indexes. Pathological histology observation by Masson and HE staining were performed for other hepatic tissues. The expression of α-smooth muscle actin (α-SMA) in hepatic tissue was analyzed by immunohistochemical technique. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP) and albumin (ALB), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and nitric oxide (NO) were determined by automatic biochemical analyzer. The levels of serum laminin (LN), type III procollagen (PCIII), type IV collagen (CIV), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured by enzyme-linked immunoassay assay (ELISA). Serum hyaluronic acid (HA) was detected by radioimmunoassay. Hepatic tissue hydroxyproline (Hyp) was measured by spectrophotometric method. Result Compared with the control group, the rats showed energielos, liver and spleen index of those were increased, as well as inflammatory cell infiltration and obvious fibrosis of hepatic tissue were appeared in the model group; The levels of ALT, AST, ALP, HA, LN, PCIII, CIV, MDA, NO, TNF-α, IL-1β and IL-6 in serum in model group were increased significantly (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD in serum were decreased significantly (P < 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly improved (P < 0.01). Compared with the model group, the levels of ALT, AST, ALP, MDA, NO in serum in NCTF group were significantly reduced (P < 0.05, 0.01); The levels of HA, LN, PC III, CIV, TNF-α, IL-1β and IL-6 were significantly reduced (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD were significantly increased (P < 0.05, 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly reduced (P < 0.05, 0.01). Compared with the model group, the pathological damage of hepatic tissue was significantly improved, the degeneration and necrosis of liver cells in rat were significantly reduced (P < 0.05). Conclusion: NCTF has a better anti-hepatic fibrosis effect, its mechanism is related to antioxidant, regulation of collagen synthesis and inhibition of the proinflammatory factors expression.