Objective To observe the protective action of cordyceps polysaccharides (CP) on photoaging skin fibroblasts. Methods Photoaging skin fibroblast models were induced by 8-MOP/UVA. Cordyceps polysaccharides was administrated before 8-MOP/UVA . HE stained, MTT, hydroxyproline (HYP), MDA and SOD quantitant were used to test the effects of cordyceps polysaccharides. Results Cell crimple, condensation of nuclear chromatin in 8-MOP/UVA model group were observed. Compared with model group, cp groups alleviate cell crimple could increase the activity of cell, decrease the content of MDA, and increase the level of HYP and the activity of SOD. Conclusion Cordyceps polysaccharides has protective effect on photoaging skin fibroblast.
--LI Hua, YE Ting-jie, LI Bo-qing, YING Xin-ping. Antagonism of Cordyceps Polysaccharides to Photoaging Fibroblasts Induced by 8-MOP/UVA. Shi Zhen Guo Yi Guo Yao. 2009; 20 (5): 1074-1076.
Cordycepin (3'-deoxyadenosine) inhibits the growth of B16-BL6 mouse melanoma cells through the stimulation of adenosine A3 receptor followed by glycogen synthase kinase-3beta activation and cyclin D1 suppression.
Cordyceps sinensis, a parasitic fungus on the larvae of Lepidoptera, has been used as a traditional Chinese medicine. We previously reported that the growth of B16-BL6 mouse melanoma (B16-BL6) cells was inhibited by cordycepin (3'-deoxyadenosine), an active ingredient of C. sinensis, and its effect was antagonized by MRS1191, a selective adenosine A3 receptor antagonist. In this study, the radioligand binding assay using [125I]-AB-MECA (a selective adenosine A3 receptor agonist) has shown that B16-BL6 cells express adenosine A3 receptors and that cordycepin binds to these receptors. We also confirmed the involvement of adenosine A3 receptors in the action of cordycepin using MRS1523 and MRS1220, specific adenosine A3 receptor antagonists. Next, indirubin, a glycogen synthase kinase-3beta (GSK-3beta) inhibitor, antagonized the growth suppression induced by cordycepin. Furthermore, the level of cyclin D1 protein in B16-BL6 cells was decreased by cordycepin using Western blot analysis. In conclusion, this study demonstrated that cordycepin inhibits the proliferation of B16-BL6 cells by stimulating adenosine A3 receptors followed by the Wnt signaling pathway, including GSK-3beta activation and cyclin D1 inhibition.
Yoshikawa N, Yamada S, Takeuchi C, Kagota S, Shinozuka K, Kunitomo M, Nakamura K. Naunyn Schmiedebergs Arch Pharmacol. 2008 Jun;377(4-6):591-5. Epub 2007 Dec 15.
Cordycepin induces apoptosis by enhancing JNK and p38 kinase activity and increasing the protein expression of Bcl-2 pro-apoptotic molecules
Objective: To explore the molecular mechanism by which cordycepin inhibits cell proliferation and induces apoptosis of human colorectal cancer cells. Methods: Cell counting and MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, inner salt) method were used to monitor the effects of cordycepin on cell proliferation. Flow cytometry (FCM) was used to analyze the effects of cordycepin on the cell cycle progress. Annexin V-fluorescein isothiocyanate (FITC) analysis was used to detect apoptosis at a very early stage. Caspase-Glo was used to determine caspase activity and Western blot was used to measure protein expression levels of c-Jun N-terminal kinase (JNK), p38, and Bcl-2 pro-apoptosis family.
Results: The numbers of viable SW480 and SW620 cells and the proliferation of these cells were significantly reduced with increases in cordycepin concentration (P<0.01). The cell cycle progression of SW480 and SW620 was arrested at the G0/G1 phase by the addition of cordycepin, and apoptosis rates of cordycepin treatments were increased compared with the control group. Cordycepin-treated cells showed phosphatidylserine valgus, suggesting the existence of early apoptosis. Caspase-3/7 and -9 activity significantly increased and the protein expression levels of JNK, p38, and Bax, Bid, Bim, and Puma from Bcl-2 pro-apoptosis molecules also increased after the treatment with cordycepin.
Conclusions: Cordycepin can inhibit SW480 and SW620 cell proliferation and induce apoptosis. Apoptosis might be induced by enhancing JNK and p38 kinase activity and increasing the protein expression of Bcl-2 pro-apoptotic molecules.
To study the effects of cordycepin on cell proliferation, we used MTS dye assay to analyze the proliferation of SW480 and SW620 cells. The results showed that the proliferation of SW480 (IC50 is 2 mmol/L) and SW620 (IC50 is 0.72 mmol/L) cells was significantly inhibited with increasing concentration of cordycepin (P<0.05 or P<0.01). Additionally, we counted the numbers of SW480 and SW620 cells after the treatment of cordycepin. The results showed that the cell numbers were significantly reduced with cordycepin in dose- and time-dependent manners (P<0.01). Our combined results imply that cordycepin inhibited the proliferation of colorectal cancer cells.
To study the molecular mechanism by which cordycepin inhibits the proliferation of colorectal cancer cells, we used FCM to analyze the effects of cordycepin on cell cycle progression and apoptosis, and we also measured the caspase enzyme activity of the cells treated with cordycepin. The results showed that the cell cycle progression was significantly inhibited in the G0/G1 phase and apoptosis rates increased significantly after the treatment with cordycepin. Annexin V assay showed early apoptosis induced by cordycepin in both SW480 and SW620 cells. Caspase enzyme activity assay showed that the increase in caspase-3/7 and -9 activity was significant, which agreed with other report (Jen et al., 2009). All of these data suggest that cordycepin arrests cell cycle progression and promotes apoptosis.
Compared with the control group, the difference was statistically significant (P<0.01).
To study the mechanism by which cordycepin inhibits cell cycle progression and induces cell apoptosis, SW480 cells were used for protein expression analysis. MAPK and Bcl-2 pro-apoptotic members were analyzed as target proteins by Western blot. The results showed that the protein expression levels of JNK and p38 kinase increased significantly after the treatment with cordycepin, while the expression level of ERK1/2 was not affected. The protein expression levels of Bcl-2 pro-apoptotic molecules (Bax, Bid, Bim, and Puma) also changed. Bid and Puma protein expression levels were positively correlated with cordycepin concentration, while the increases of Bid and Puma protein levels were more obvious in lower-dose cordycepin-treated cells. Together, these results imply that cordycepin induced cell apoptosis through activation of JNK and p38 kinase pathways and increases of the expression levels of Bcl-2 pro-apoptotic members such as Bim, Bid, Bax, and Puma.
He W, Zhang M-f, Ye J, et al. J Zhejiang Univ Sci B. 2010 September; 11(9): 654–660.