Immuno-enhancement effects of ethanol extract from Cyrtomium macrophyllum (Makino) Tagawa on cyclophosphamide-induced immunosuppression in BALB/c mice

Abstract

Ethnopharmacological relevance

Cyrtomium macrophyllum (Makino) Tagawa has been traditionally used as a herbal medicine for the treatment of various infectious diseases such as tapeworm infestation, colds, and viral diseases. However, no systematic study of the immunity of Cyrtomium macrophyllum ethanol extracts (CM) has yet been reported. The present work evaluates these traits.

Materials and methods

120 male BALB/c mice were divided into 6 groups of 20 mice each: (1) normal group (sterile physiological saline), which served as a blank control; (2) model group (Cyclophosphamide, CY) group (sterile physiological saline), which served as a negative control; (3) low-dose CM (50 mg/kg BW); (4) intermediate-dose CM (100 mg/kg BW); (5) high-dose CM (200 mg/kg BW); (6) CM group (200 mg/kg BW). CY (0.2 ml) was administered via intraperitoneal injection. The other regimens were administered via gavage in 0.2 ml solution. Phytochemical of CM was characterized by HPLC–LTQ-Orbitrap. The acute toxicity effect of the ethanol extract of Cyrtomium macrophyllum was also investigated.

Results

The spleen and thymus indices of mice receiving low, intermediate, and high doses of CM recovered more quickly than those of CY mice, and they did so in a dose-dependent manner. These mice also showed higher T cell and B cell proliferation responses and macrophage function than those of CY mice, and their serum levels of interleukin-6 and interferon-γ had become normal. In acute toxicity test, CM exhibited no mortality and behavioral changes in mice. Quantitative phytochemical analysis showed flavonoids, polyphenols, and tannins to be the major compounds present in the extract, at 27.64%, 30.87%, and 11.22%, respectively. We found that 16 compounds were characterized by the interpretation of their mass spectra obtained by the MS/MS.

Conclusion

The current study demonstrates that Cyrtomium macrophyllum ethanol extract improved immune function in CY-treated mice.

Introduction

A large number of plants used in traditional medicines have been shown to possess immunomodulating activities (Choi et al., 2004, Ben Sghaier et al., 2011). Recently, complementary or alternative medicines have become popular for treating different immune disorders. Increasingly among these are extracts from medicinal plants. Evaluation of the immunomodulatory activity of plant extracts is an interesting and growing area of research. However, little is known about its immunomodulatory activities, in particular, for less frequently studied or previously unknown (medically) plants.

Cyrtomium macrophyllum (Makino) Tagawa f. muticum (Christ) Ching et Shing (Family: Dryopteridaceae) is a common medicinal plant found in the southwest of China. It is recorded as Daye Guanzhong in Traditional Chinese Medicines (Wu and Song, 1996). Decoctions of Cyrtomium macrophyllum are used in folk medicine for the treatment of a variety of diseases including, epidemic flu, epidemic encephalitis B, homeostasis and dysfunctional uterine bleeding (Wu and Song, 1996, Lou and Qin, 2003, Liu et al., 2004) and their efficacy is widely acclaimed among the Guizhou Province of China. Previous studies suggest that the rhizome of Cyrtomium may contain various pharmacological actions such as anti-oxidative, anti-bacterial, anti-parasite, anti-viral and anti-cancer (Yang and Li, 1993, Kapadia et al., 1996, Ito et al., 2000, Zhou et al., 2009, Da Silva et al., 2010, Cao et al., 2013).

These activities are mediated by active components such as flavonoids, phloroglucinol derivatives, triterpenes, tannins, essential oils and polysaccharides (Hisada et al., 1972). Major flavonoids were flavonol O-glycosides based on kaempferol, quercetin, and sometimes myricetin, and C-glycosylflavones, such as isovitexin, vitexin, isoorientin, orientin and their O-glycosides (Xiang et al., 2002, Iwashina et al., 2006).

However, there is too little information on the components and bioactivities of most fern species including Cyrtomium macrophyllum. In this study, we aimed to demonstrate in vivo ethnomedicinal benefits of ethanolic extract from Cyrtomium macrophyllum against immunosuppressive conditions. The major compounds in the tested extracts were tentatively identified by high performance liquid chromatography–linear ion trap-Orbitrap hybrid mass spectrometry (HPLC–LTQ-Orbitrap) analysis for the first time for the rhizome of this species. In particular, this is the first report of immunomodulatory activities of this plant collected from Guizhou province.