Free radical scavenging and apoptotic effects of Cordyceps sinensis fractionated by supercritical carbon dioxide
Introduction
In order to develop an anti-tumor and anti-radical agent with low toxicity on normal cells, we assayed a number of natural products that have been used to treat diseases for hundreds of years and have been proven to be nontoxic. Cordyceps sinensis (CS), a parasitic fungus on the larvae of Lepidoptera, has been used as one of the most valued traditional Chinese herbal remedy for centuries to maintain health (Nakamura et al., 1999). The biological significance of CS includes its regulations on glucose and lipid metabolism, endocrine secretion, sexual function, free radical clearance and immunity (Zhu et al., 1998a, Zhu et al., 1998b).
The isolation of active constituents using a nonhazardous protocol is important to ensure the safety of the administration of anti-cancer and anti-radical agents. Supercritical carbon dioxide (SC-CO2) was used herein as the fraction solvent (Wang et al., 2004) to solve this problem. The constituents of extracts differ with the extraction methods; for example, the supercritical extract of propolis contains more diterpens and less flavonoids than ethanol extracts (Matsuno et al., 1977). Fractionation is a separation method to minimize the co-extraction of undesired compounds and to yield fractions with different compositions (Yang and Wang, 1999). We have reported that SC-CO2 extractive fractionation successfully fractionated the ethanol extract of Bupleurum kaoi into four fractions (R, F1, F2, and F3) (Wang et al., 2004). Among these fractions, F3 contained more phenol compounds and thus exhibited the most antioxidant capability, but R contained more saikosaponins and thus was the best fraction to protect the liver against the damage caused by CCl4. According to our preliminary experiments, ethanol extract of CS exhibited the strongest growth inhibition and antioxidant activities compared with those extracted with solvents of different polarity. The incidence of tumors increases with exposure to free radicals (Hussain et al., 2003). Anti-oxidative capacity for scavenging free radicals was responsible for its anti-mutagenic effect (Krizkova et al., 2003). Various constituents such as nucleosides and polysaccharides are associated with the pharmacological activities of Cordyceps sp. (Frederikson and Klenow, 1975; Shin et al., 2001). Therefore, in the present study, the ethanol extract was further fractionated with SC-CO2, and the anti-tumor and anti-radical abilities of the ethanol extract and its fractions were determined. The concentrations of polysaccharides and cordycepin in each fraction were also addressed. Human colorectal and hepatocellular carcinoma cells were chosen for the study, since they are globally common malignant tumors, and are responsible for significant mortality in both Western countries and Taiwan due to their high recurrence rate and poor prognosis (Di Bisceglie et al., 1988; Landis et al., 1999).
Apoptosis modulates cell cycle and finally leads to cell death (Paulovich et al., 1997). Unlike necrosis in which dying cells fall apart releasing their contents and provoking macrophage activation and inflammation, apoptotic cells are rapidly taken up by phagocytosis and degraded in phagolysomes (Roitt et al., 2001). Various tumors undergoing the process of apoptosis were induced by treatments of radiation, chemotherapeutic agents and mild hyperthermia (Kerr et al., 1994). Our results indicated that CS fraction induced apoptosis in cancer cells.
Section snippets
Materials
Culture mycelium of Cordyceps sinensis (CS) was from Green Health Biotechnology Corporation, Yunlin, Taiwan. Human hepatocellular cells, Hep G2 and Hep 3B, and colorectal carcinoma cells, HT-29 and HCT 116, were from the American Type Culture Collection (ATCC, Rockville, MD). Human peripheral blood mononuclear cells (PBMC) were obtained from freshly collected buffy coat fraction from healthy donors at the Tainan Blood Bank Center (Tainan City, Taiwan, ROC). Dulbecco’s modified Eagle medium
DPPH free radical scavenging activity
Fig. 1 displayed the scavenging effect of CS extract and fractions on free radicals. The absorbance at 517 nm decreases when DPPH accepts an electron or hydrogen radical. DPPH radical scavenging ability increased with the concentrations of CS extract and fractions. Of these fractions, R was the most active fraction for scavenging free radicals. The scavenging effect of CS extract and fractions on DPPH at 2 mg/ml was 93% (R), 75% (F1), 66% (E), 47% (F2), and 27% (F3), respectively. The free
Discussion
The present study aimed to investigate the anti-tumor and anti-radical effects of CS extract and its fractions prepared by SC-CO2. The selectivity of fractionation with SC-CO2 is based on the change in CO2 density with total pressure and the operating temperature of the separation system. The present experimental results indicated that fractionation altered the composition distributions of fractions, and the fraction R exhibited the best activity against DPPH free radicals and triggered the
Acknowledgments
The authors would like to thank Green Health Biotechnology Corporation, Yunlin, Taiwan and the National Science Council of Taiwan, Republic of China for financially supporting this research under contract no. NSC 89-2316-B-309-001.
References (47)
- et al.
Effect of Cordyceps sinensis on the proliferation and differentiation of human leukemic U937 cells
Life Science
(1997) - et al.
Antiherpetic activities of various protein bound polysaccharides isolated from Ganoderma lucidium
Journal of Ethnopharmacology
(1999) - et al.
A comparison of the chemical composition and bioactive ingredients of the Chinese medicinal mushroom DongChongXiaCao, its counterfeit and mimic, and fermented mycelium of Cordyceps sisnensis
Food Chemistry
(2002) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
Journal of Immunological Methods
(1983)- et al.
When checkpoints fail
Cell
(1997) - et al.
Glucans exhibit weak antioxidant activity, but stimulate macrophage free radical activity
Free Radical Biology and Medicine
(2001) - et al.
Hepatoprotective and antioxidant effects of Bupleurum kaoi Liu (Chao et Chung) extract and its fractions fractionated using supercritical CO2 on CCl4-induced liver damage
Food and Chemical Toxicology
(2004) - et al.
The supercritical fluid extractive fractionation and the characterization of heavy oils and petroleum residua
Journal of Petroleum Science and Engineering
(1999) - et al.
H1-A extracted from Cordyceps sinensis suppresses the proliferation of human mesangial cells and promotes apoptosis, probably by inhibiting the tyrosine phosphorylation of Bcl-2 and Bcl-XL
Journal of Laboratory and Clinical Medicine
(2003) - et al.
Positive correlation between histological tumor involvement and generation of tumor cell colonies in agarose in specimens taken directly form patients with small-cell carcinoma of the lung
Cancer Research
(1980)
NIH conference. Hepatocellular carcinoma
Annals of Internal Medicine
DNA damage triggers a prolonged p53-dependent G1 arrest and long-term induction of Cip1in normal fibroblasts
Genes and Development
Antineoplastic and Immunosuppressive Agent II
Effects of the interaction of tannins with co-existing substances. VI. Effects of tannins and related polyphenols on superoxide anion radical, and on 1,1-diphenyl-perylhydrazyl radical
Chemical Pharmaceutical Bulletin
Effects of extracted Cordyceps sinensis on steroidogenesis in MA-10 mouse Leydig tumor cells
Biological and Pharmaceutical Bulletin
Radical causes of cancer
Nature reviews Cancer
Apoptosis. Its significance in cancer and cancer therapy
Cancer
Polysaccharides in fungi. XXXVI. Hypoglycemic activity of a polysaccharide (CS-F30) from the cultural mycelium of Cordyceps sinensis and its effect on glucose metabolism in mouse liver
Biological and Pharmaceutical Bulletin
Antitumor activity of extract of Zingiber aromaticum and its bioactive sesquiterpenoid zerumbone
Nutrition and Cancer
Induction of apoptosis by cordycepin in ASA-inhibited TdT-postive leukemia cells
Leukemia
Hypocholesterolemic effect of hot-water extract from mycelia of Cordyceps sinensis
Biological and Pharmaceutical Bulletin
Funal beta-(1-3)-d-glucan derivatives exhibit high antioxidative and antimutagenic activity in vivo
Anti-cancer Research
Cited by (119)
Phenolic content and antioxidant activity of Ophiocordyceps Sobolifera extract for renal injury prevention
2022, Process BiochemistryCitation Excerpt :Having beneficial effects on human health, some species of Cordyceps spp. have been used as traditional medicine for immunity modulation, fatigue resistance, longevity, and other treatments [3–6]. Modern medical studies have reported various pharmacological activities of Cordyceps, such as anti-tumor, antioxidant, anti-aging, hyposexuality, and hyperlipidemia, as well as valuable effects for the treatments of respiratory, renal, hepatic, neurological, and cardiovascular diseases [7–9,4,10–13]. These properties of Cordyceps spp. are attributed to their chemical components, mainly including polysaccharides, cordycepin, adenosine, ergosterol, cordycepic acid, phenolic, flavonoid, etc. [14,15,3,16–18].
Leukemia cells apoptosis by a newly discovered heterogeneous polysaccharide from Angelica sinensis (Oliv.) Diels
2020, Carbohydrate PolymersCitation Excerpt :Polysaccharide is a major ingredient in Angelica sinensis with many bioactivities, including antioxidant, antitumor, antiaging, antihepatotoxic, immunomodulatory, and neuroprotective effects (Cao et al., 2006; Jin, Zhao, Huang, Xu, & Shang, 2012). Studies have showed that Angelica polysaccharides could significantly inhibit the proliferation of leukemia cells and induce their apoptosis (Liu et al., 2019; Wang et al., 2005). In addition, Angelica polysaccharides significantly promoted the spleen and bone marrow hematopoietic function (Cao, Li, Wang, Li et al., 2010, Cao, Li, Wang, Fan, 2010).
Polysaccharides from Pyracantha fortuneana and its biological activity
2020, International Journal of Biological MacromoleculesThe novel indole glucoalkaloid and secoiridoid glucoside from Tripterospermum chinense
2020, Phytochemistry LettersCitation Excerpt :The plate was incubated for 4 days at 37 °C in a humidified, 5 % CO2 atmosphere. Cytotoxicity was determined using a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay (Wang et al., 2005). After addition of 10 μL MTT solution (5 mg/mL), cells were incubated at 37 °C for 4 h.
Extraction and characterization of an alginate from the Iranian brown seaweed Nizimuddinia zanardini
2018, International Journal of Biological Macromolecules