Beauvericin (BEA) is a cyclohexadepsipeptide produced by the fungus Beauveria bassiana. And it has been reported to have very effective anti-cancer activity. However, the mechanistic studies of BEA on cytotoxicity on cancer cells have not been fully understood. In this study, we examined the cytotoxicity of BEA on the C6 (rat glioma cell line). The cell viabilities of C6 cells, MDA-MB 231 (breast cancer cell line), and HeLa (cervical cancer cell line) were decreased by treatment of BEA. In addition, BEA-treated cells showed membrane blebbings and buddings, which indicates that BEA decreased cell viability by inducing apoptosis. Additionally, we found that the level of apoptosis inducing molecules such as caspase 3, caspase 9 of BEA treated cells was increased and apoptosis regulating molecules (Bcl-2) was decreased. We also found that the activated STAT3 and Src was decreased in BEA-treated condition. In further study, we are going to find out how the BEA can influence Src kinase activity.

The inhibitory activities of the Cordyceps pruinosa butanol fraction (Cp-BF) were investigated by determining inflammatory responses of lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells and by evaluating HCl/ethanol (EtOH)-triggered gastric ulcers in mice. The molecular mechanisms of the inhibitory effects of Cp-BF were investigated by identifying target enzymes using biochemical and molecular biological approaches. Cp-BF strongly inhibited the production of NO and TNF-α, release of reactive oxygen species (ROS), phagocytic uptake of FITC-dextran, and mRNA expression levels of interleukin (IL)-6, inducible NO synthase (iNOS), and tumour necrosis factor-alpha (TNF)-α in activated RAW264.7 cells. Cp-BF also strongly down regulated the NF-κB pathway by suppressing IKKβ according to luciferase reporter assays and immunoblot analysis. Furthermore, Cp-BF blocked both increased levels of NF-κB-mediated luciferase activities and phosphorylation of p65/p50 observed by IKKβ overexpression. Finally, orally administered Cp-BF was found to attenuate gastric ulcer and block the phosphorylation of IκBα induced by HCl/EtOH. Therefore, these results suggest that the anti-inflammatory activity of Cp-BF may be mediated by suppression of IKKα and its downstream NF-κB activation. Since our group has established the mass cultivation conditions by developing culture conditions for Cordyceps pruinosa, the information presented in this study may be useful for developing new anti-inflammatory agents.

4-isopropyl-2,6-bis(1-phenylethyl)aniline 1 (KTH13-AD1) is a metabolite of Cordyceps bassiana that has been traditionally used to treat various inflammatory disease. Even though it has the magnificent pharmaceutical potential, there was not much understanding on its anti-inflammatory actions. Therefore, this study was aimed to determine the anti-inflammatory effects of KTH13-AD1. We found that KTH13-AD1 suppressed nitric oxide (NO) and reactive oxygen species (ROS) production in lipopolysaccharide (LPS)- or sodium nitroprusside (SNP)-treated macrophages (RAW264.7 cells). Similarly, mRNA expression of inducible NO synthase (iNOS) and tumor necrosis factor-a (TNF-a) analyzed by RT-PCR and real-time PCR was also reduced by this compound. Interestingly, KTH13-AD1 also strongly diminished the levels of NF-kB-mediated luciferase activities and nuclear translocated NF-kB family proteins. In agreement with these, KTH13-AD1 suppressed the upstream signaling pathway for NF-kB activation including IkBa, IKKa/b, AKT, p85/PI3K and Src in time- and dose-dependent manners. Therefore, these results strongly suggest that KTH13-AD1 has a strong anti-inflammatory activity via suppression of the NF-kB signaling pathway.

In this study, we aimed to examine the cellular and molecular mechanisms of KTH-13 (4-isopropyl-2,6-bis(1-phenylethyl)phenol) which is derived from Cordyceps staphylindaecola in the cancer cells survival. The apoptotic effect of KTH-13 on various cancer cells, such as C6 glioma, MDA-MB-231 breast cancer, and A549 cells, was determined by MTT assay, and result showed that KTH-13 (0-100mM) dramatically inhibited the cancer cell survival. IC50 of KTH13 were 60.549, 53.512, >100, in C6, MDA-MB-231, and A549, respectively. DNA fragmentation result revealed that MDA-MB-231 cells treated with KTH-13 100mM undergoes apoptosis. To understand the action mechanism of KTH-13, the effect of KTH-13 on caspase which is key regulator of apoptosis was verified. The amount of cleaved capspase-3 and 7, executioner caspases, was increased by KTH-13 treatment, at time dependent (capspase-3 case) and dose dependent manner (caspase-7 case). And the cleavage of caspase-9 which is initiator caspase was also elevated in KTH-13 treated MDA-MB-231 cells showing time dependent manner. However, caspase-8 was not regulated by KTH-13, indicating KTH-13 specifically targets caspase-9 signal. As caspase-9 is closely associated with intrinsic pathway, the involvement of bcl-2 family was identified. Bax, pro-apoptotic molecule, was up-regulated whereas Bcl-2, anti-apoptotic protein, was down-regulated. And the Bax/Bcl-2 ratio was increased about 10 times. Then, the survival signal was also observed. The phosphorylation of Akt and p85 was diminished by KTH-13 treatment at 2,4,6 and 8 hour. Collectively, results suggest that KTH-13 induces cancer cells apoptosis via caspase3, 7, 8 and Bcl-2 family signaling pathway. And the Akt and p85 is also involved in KTH-13 action mechanism.