Extracts Characterization and In Vitro Evaluation of Potential Immunomodulatory Activities of the Moss Hypnum cupressiforme Hedw

Abstract

Recently, there has been an increasing interest in the chemistry and biological potential of mosses, since a large number of biologically active compounds have been found within these species. This study aimed at examining the chemical composition and immunomodulatory potential (antioxidant, antidiabetic, anti-neuroinflammatory/antineurodegenerative, and antitumor activities) of moss Hypnum cupressiforme Hedw. extracts. Corresponding extracts have been obtained applying Soxhlet extractor. The chemical characterization was performed using spectrophotometric assays and liquid chromatography-mass spectrometry (LC-MS). The extracts were analyzed for antioxidant activity and for inhibitory activities on α-glucosidase, α-amylase, acetylcholinesterase, and tyrosinase. Additionally, extracts were tested against four cell lines-MRC-5, BV2, HCT-116, and MDA-MB-231-for antitumor and anti-inflammatory activities. Chemical analysis of extracts revealed the presence of flavonoids, phenolic acids, and triterpenoids. Major compounds identified by LC-MS in H. cupressiforme were kaempferol and five phenolic acids: p-hydroxybenzoic, protocatechuic, p-coumaric, gallic, and caffeic acid. According to biochemical assays the investigated extracts exhibited significant immunomodulatory potential. Significant antiproliferative potential against MDA-MB-231 cells has been observed together with the promising anti-neuroinflammatory application. The obtained data suggest that moss H. cupressiforme is a valuable natural source of biologically active compounds with potential application in the pharmaceutical industry.

Keywords: Hypnum cupressiforme; anti-neuroinflammatory/antineurodegenerative; antidiabetic; antioxidant; antitumor activity; moss extract.

Figure 1

Biochemical analysis of Hypnum cupressiforme extracts as (A) antioxidant and (B) antidiabetic agents. The results are expressed as the mean ± standard error from an experiment performed in triplicate (#, +, * p < 0.05 different moss extracts vs. different standard substances. Symbols # and + were used for standards BHA and AA (ascorbic acid), while * was used for all the other standards).

Figure 2

Biochemical analysis of Hypnum cupressiforme extracts as (A) antiacetylcholinesterase and (B) antityrosinase agents. The results are expressed as the mean ± standard error from an experiment performed in triplicate (* p < 0.05 different moss extracts vs. different standard substances).

Figure 3

Immunomodulatory potential of Hypnum cupressiforme extracts (final concentration 10 µg/mL) on (A) cell proliferation, (B) ROS production, and (C) NO production. The results are expressed as mean ± standard error of a representative experiment performed in quadruplicate. (* p < 0.05 different moss extracts and LPS-stimulated control cells vs. non-stimulated control cells; ** p < 0.05 different moss extracts vs. only LPS-stimulated control cells).

Figure 4

The position of the investigated collection area (Vršačke Planine Mts.) in Serbia with illustrated steps in the preparation and characterization of the extracts.