Abstract

BackgroundChrysanthemum indicum L. flower (CIF) has been widely used as tea in Korea. This study aims to investigate the hepatoprotective effect of the hot water extract of CIF (HCIF) in in vitro and in vivo systems.MethodsHepatoprotective activities were evaluated at 250 to 1000 μg/mL concentrations by an in vitro assay using normal human hepatocytes (Chang cell) and hepatocellular carcinoma cells (HepG2) against CCl4-induced cytotoxicity. Cytochrome P450 2E1, which is a key indicator of hepatic injury, was detected by western blot analysis using rabbit polyclonal anti-human CYP2E1 antibody. An in vivo hepatoprotective activity assay was performed at 1000 to 4000 μg/mL concentrations on CCl4-induced acute toxicity in rats, and the serum levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were determined by standard enzyme assays.ResultsThe hepatoprotective effects of HCIF significantly reduced the levels of GOT (60.1%, P = 0.000) and GPT (64.5%, P = 0.000) compared with the vehicle control group (CCl4 alone). The survival rates of HepG2 and Chang cells were significantly improved compared with the control group [82.1% (P = 0.034) and 62.3% (P = 0.002), respectively]. HCIF [50 mg/kg body weight (BW)] treatment significantly reduced the serum levels of GOT (49.5%, P = 0.00), GPT (55.5%, P = 0.00), ALP (30.8%, P = 0.000) and LDH (45.6%, P = 0.000) compared with the control group in this in vivo study. The expression level of cytochrome P450 2E1 (CYP2E1) protein was also significantly decreased at the same concentration (50 mg/kg BW; P = 0.018).ConclusionHCIF inhibited bioactivation of CCl4-induced hepatotoxicity and downregulates CYP2E1 expression in vitro and in vivo.

Highlights

  • Chrysanthemum indicum L. flower (CIF) has been widely used as tea in Korea

  • Chemicals and reagents Bovine serum albumin (BSA), 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), sodium bicarbonate, silymarin and Carbon tetrachloride (CCl4) were purchased from Sigma Chemical Co

  • Effect of hot water extract of CIF (HCIF) on CCl4-induced hepatotoxicity in vitro The 8 mM CCl4-exposed Hepatocellular carcinoma cell line (HepG2) and Chang cells exhibited cell viabilities of 58% and 39%, respectively, compared with untreated controls (Figure 1). Viability of these CCl4-exposed cells was exhibited in a dosedependent manner when pretreated with various HCIF concentrations

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Summary

Introduction

Chrysanthemum indicum L. flower (CIF) has been widely used as tea in Korea. CCl4 is a widely used chemical and causes severe liver tissue damage by undergoing biotransformation by the cytochrome P450 system into a trichloromethyl free radical (CCl3) and transformation into a highly reactive trichloromethylperoxy free radical (CCl3O2). Chrysanthemum indicum L. flower (CIF) is a wild herb and has a long history of use as a traditional medicine, mainly for the treatment of inflammation, hypertension and respiratory diseases in Korean and Chinese medicine [6,7,8,9]. Several studies have demonstrated that the water extract of C. indicum L. has strong antioxidant effects and inhibitory effects against bacteria and viruses [10,11]. Several chemical compounds isolated from CIF exhibit inhibitory activity against nitric oxide (NO) in lipopolysaccharide-activated macrophages and rat lens aldose reductase [13]

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