본 연구에서는 고분자 점도 조절제를 첨가하여 졸-겔법 기반 알루미나 나노여과막을 단일 공정으로 제조하고, 코 팅층의 구조 및 성능을 제어하는 방법을 제시하였다. Hydroxypropyl cellulose (HPC, Mw ~80000) 고분자를 알루미나 졸에 첨가하여 점도를 10 mPa·s에서 최대 4200 mPa·s까지 조절하였으며, 이를 통해 알루미나 중공사 지지체 표면에 균일하고 결 함이 없는 선택층을 형성하였다. HPC 함량이 증가할수록 코팅층 두께가 증가하였으나, 기공 크기 증가에 따라 분리 성능이 저하되었다. 2:1 (졸:HPC 고분자 용액) 혼합비에서 제조된 나노여과막은 두께 3.20 μm의 얇은 선택층을 형성하여 높은 수투 과도(12.9 LMH/bar)와 우수한 제거 성능(PPG 1050 Da 제거율 60%, PEG 1500 Da 제거율 90%, MgCl2 제거율 80%)을 나타 냈다. 반면, 1:2 혼합비에서는 선택층 두께가 10.2 μm로 증가하였으나, 기공 크기가 증가하여 3400 Da MWCO와 64% 염 제 거율을 보였다. HPC 고분자를 활용한 점도 제어는 졸-겔 코팅층의 두께, 기공 구조 및 분리 성능을 효과적으로 조절할 수 있 음을 입증하였다.

Cordycepin, a specific polyadenylation inhibitor, is the main functional component in Cordyceps militaris that is one of the top three renowned traditional Chinese medicines. In this study, we performed in vitro experiments to investigate the anti-invasive and anti-metastatic activities of cordycepin using human prostate carcinoma LNCaP cells. Cordycepin were administered and their effects on LPS-induced cell migration and invasion by wound healing migration assay, measurement of TER and In vitro invasiveness assay. Within the concentrations which were not cytotoxic effects, cordycepin caused a concentration-dependent suppression of LPS-induced cell migration and invasion. The anti-invasive activity of cordycepin was also found to be associated with increased tightness of the TJ, which was confirmed by an increase in TER. The activity of MMP-2 in LNCaP cells was dose-dependently inhibited by treatment with cordycepin, and this was also correlated with a decrease in expression of its mRNA and proteins, and up-regulation of TIMPs expression. Additionally, cordycepin repressed the LPS-induced NF-kB activation and phosphorylation of PI3K/AKT. Taken together, these findings suggest that cordycepin inhibited LPS-induced migration and invasion of LNCaP cells by down-regulating the expression and activity of MMP-2, and the possible targets may be NF-kB and PI3K/AKT.

[Background] Cordyceps militaris is a traditional popular mushroom, produces an important bioactive compound Cordycepin (3’-deoxyadenosine) used for the tonic and medicinal purpose in eastern Asia. Cordycepin is reported to possess many pharmacological activities including immunologically stimulating, anti-tumor, anti-virus, and anti-infection effects. [Methods] Growth inhibition of human leukemia cells was assessed by MTT assays. The determination of apoptotic cell death was performed by flow cytometry analysis, agarose gel electrophoresis and DAPI fluorescent staining methods. The apoptotic-regulated gene markers in both death receptor- and mitochondria-mediated apoptotic pathways were detected by RT-PCR and Western blot analysis etc. [Results] It was found that inhibition of cell proliferation was observed for human leukemia U937 and THP-1 cells treated with cordycepin in a dose-dependent manner. Cordycepin induced morphological change and apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptotic-sub G1 phase. Apoptosis of U937 and THP-1 cells by cordycepin was associated with a down-regulation of anti-apoptotic Bcl-2 and inhibitor of apoptosis proteins (IAPs) expression. Cordycepin treatment induced the proteolytic activation of caspase-3, caspase-8 and caspase-9, and a concomitant inhibition of poly(ADP-ribose) polymerase (PARP), β-catenin and phospholipase (PLC)-γ1 protein. Conclusions: Our results indicated that the apoptotic processes caused by cordycepin are mediated by the regulation of the Bcl-2 and caspase family in human leukemia U937 and THP-1 cells. Our data also suggested that cordycepin may be a potential chemotherapeutic agent for the treatment of leukemia cancer patients.

[Background] Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3’-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. However, the molecular mechanisms of inflammatory mediator’s activity by cordycepin remain poorly understood. In the present study, we investigat-ed the effects of cordycepin on the anti- inflammation cascades in lipopolysaccharide (LPS)-stimulated BV2 microglia cells. [Methods] Cordycepin were administered and their effects on LPS-induced pro-inflammatory mediators and MAP kinases were monitored by Western blotting and RT-PCR analysis. [Result] Cordycepin significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), and pro- inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in a concentration- dependent manner without causing cytotoxicity. Also, cordycepin suppressed inducible NO, synthase (iNOS) and cyclooxygenase-2 (COX-2) expression on the mRNA and protein level. In addition, cordycepin suppressed NF-κB translocation by blocking IkappaB- α (IκB-α) degradation and inhibited the phosphorylation of Akt, ERK-1/2, JNK, and p38 kinase. Our results indicate that the inhibitory effect of cordycepin on LPS -stimulated inflammatory mediator production in BV2 microglia is associated with the suppression of the NF-κB, Akt, and MAPK signaling pathways. Conclusion: Anti-inflammatory properties of cordycepin may be useful for treating the inflammatory and deleterious effects of microglial activation in response to LPS stimulation.

Cordycepin (3’-deoxyadenosin), a polyadenylation specific inhibitor, is the main functional component in Cordyceps militaris which is one of the top three famous traditional Chinese medicine. It has been shown to possess many pharmacological activities including immunologically stimulating, anti-cancer, anti-bacterial, and anti-virus, anti-infection effects. However, its anti-cancer molecular mechanisms are poorly understood. In this study, the apoptotic effects by cordycepin were investigates in human leukemia cells. Treatment of cordycepin significantly inhibited cells growth in a concentrationdependent manner by inducing apoptosis, as evidenced by morphological change and apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of sub-G1. Induction of apoptosis by cordycepin was associated with modulation of Bcl-2 and inhibitor of apoptosis proteins (IAP) family expression. Cordycepin also increased reactive oxygen species (ROS) generation, activation of casepase-3, caspase-8, caspase-9, cleavage of poly(ADP-ribose) polymerase (PARP), β-catenin and phospholipase C (PLC)-γ1 protein. The quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the cordycepin-induced apoptosis effects. Theresults suggested that cordycepin may be a potential chemotherapeutic agent for the treatment of leukemia patients [This work was supported by Blue-Bio Industry RIC at Dong-Eui University as a RIC (08-06-07) program of ITEP under Ministry of Knowledge Economy].

Cordycepin (3’-deoxyadenosine) is a polyadenylation specific inhibitor, one of the components of Cordyceps militaris. It has been shown to possess many pharmacological activities including immunologically stimulating, anti-tumor, anti-virus, and anti-infection effects. However, its molecular mechanisms are poorly understood. In this study, the apoptotic effects by cordycepin were investigated in human leukemia cells. Cordycepin treatment inhibited leukemia cells growth a concentration-dependent manner by inducing apoptosis,as evidenced by morphological change and apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptoticsub G1 phase. Induction of apoptosis by cordycepin in leukemia cells were associated with modulation of Bcl-2 member and inhibitor of apoptosis (IAP) proteins expression. Cordycepin also increased ROS generation, activation of caspase-3, caspase-8, caspase-9, cleavage of poly(ADP-ribose) polymerase (PARP), -catenin and phospholipase (PLC)-1 protein. Both the this effect by cordycepin treatment were significantly inhibited by NAC, a ROS scavenger, demonstrating the important role of ROS in the observed cytotoxic effect. This results suggested that cordycepin may be a potential chemotherapeutic agent for the treatment of leukemia patients.

Cordyceps militarisis well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militarisextract(CME) in human breast cancer MDA-MB-231 cells. It was found that CME treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNAcontent. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CME treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor,demonstrating the important role of caspase-3 in the observed cytotoxic effect. Cotreatment of CME and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CME in human breast cancer MDAMB- 231 through downregulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.

Agaricus blazei is well known as a traditional medicinal mushroom and it has been shown to exhibit immunostimulatory and anti-cancer activity. However, the cellular and molecular mechanism of apoptosis of cancer cells is poorly understood. In this study, we have investigated whether A. blazei extract (ABE) exerts anti-proliferative and apoptotic effects on human leukemia THP-1 cells. It was found that ABE induced a time- and dose-dependent increase in leukemia cells apoptosis through caspase-3 activation and PARP cleavage. Activation of caspase- 9 induced by ABE suggested that ABE-induced signaling was mediated through a mitochondrial death pathway. In addition, we observed an elevation of ROS and a consequent loss of mitochondrial membrane potential, further suggesting that ABE-induced death signaling was mediated through a mitochondrial oxygen stress pathway. The antioxidant Nacetylcysteine, however, opposed ABE-mediated mitochondrial dysfunction, caspase activation, and apoptosis, supporting the role of ROS in the apoptotic process. We conclude that ABE induces apoptosisin human leukemia cells through a reactive oxygen species and caspase-dependent mitochondrial pathway.