Dibenzylbutyrolactone lignans from Forsythia koreana fruits attenuate lipopolysaccharide-induced inducible nitric oxide synthetase and cyclooxygenase-2 expressions through activation of nuclear factor-κb and mitogen-activated protein kinase in RAW264.7 cells.

Previously, we reported that dibenzylbutyrolactone lignans (DBLLs) from the fruit of Forsythia koreana NAKAI (Oleaceae) has anti-inflammatory, antioxidant, and anti-asthmatic effects. In this study, to clarify the anti-inflammatory mechanisms of DBLL, we evaluated the effects of DBLLs on lipopolysaccharide-stimulated inducible nitric oxide synthetase (iNOS) and cyclooxygenase-2 (COX-2) expressions, nitric oxide (NO) and prostaglandin E(2) (PGE(2)) productions, nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) activations, inhibitor of κB (IκB) and inhibitor of κB kinase (IKK) phosphorylations in cytosolic proteins, and cytotoxicity in Raw264.7 cells. DBLLs potently suppressed both the enzyme expression and DNA-binding activity of NF-κB. Arctiin, arctigenin (1.0 µM) and matairesinol (10 µM) inhibited the expression of iNOS by 37.71±2.86%, 32.51±4.28%, and 27.44±2.65%, respectively, and arctiin, arctigenin (0.1 µM) and matairesinol (1.0 µM) inhibited COX-2 expression by 37.93±7.81%, 26.70±4.61% and 29.37±5.21%, respectively. The inhibitory effects of DBLLs on NO and PGE(2) productions were the same patterns as those seen for the reductions in iNOS and COX-2 expression, respectively. Arctiin, arctigenin (1.0 µM) and matairesinol (10 µM) significantly (p<0.05) inhibited NF-κB DNA binding by 44.85±6.67%, 44.16±6.61%, and 44.79±5.62%, respectively, and arctiin (0.1 µM) and arctigenin (1.0 µM) significantly (p<0.05) inhibited the phosphorylation of IκB by 20.58±3.86% and 25.99±6.18%, respectively. Furthermore, arctiin, matairesinol (1.0 µM) and arctigenin (10 µM) inhibited the phosphorylation of IKK by 38.80±6.64%, 38.33±6.65%, and 38.57±8.14%, respectively. In addition, DBLLs potently inhibited the lipopolysaccharide (LPS)-induced activation of MAPKs (SAPK/c-Jun NH(2)-terminal kinase (JNK), p38, and extracellular signal receptor-activated kinase (ERK)1/2). Overall, arctiin was the most effective; its effect was nearly the same as that of 10 µM helenalin. These findings suggest that treatment with non-toxic DBLLs inhibits not only NF-κB and NF-κB-regulated protein activation, but also potently inhibits the activations of specific MAPKs.

Effect of coenzyme Q10 on cutaneous healing in skin-incised mice.

Coenzyme Q10 (CoQ10) is a biosynthesized quinone with 10 isoprene side chains in humans. To investigate the anti-inflammatory and wound healing effect of CoQ10, we performed in vivo and in vitro experiments. In vivo studies, there were 3 groups; Naive (without skin incision), Control (with skin incision) and CoQ10 (100 mg/kg treatment with skin incision). Collagen-like polymer (CLP) level of CoQ10 group was increased significantly compared to the control group (p<0.05). Also, CoQ10 group showed significant inhibition on myeloperoxidase (MPO) and PLA(2) level compared to the control group (p<0.05). These data show that CoQ10 may have an anti-inflammatory and a wound healing effect. CoQ10 showed significant antioxidant activity in vivo on malondialdehyde (MDA) and superoxide dismutase (SOD) levels compared to the control group (p<0.05). Although CoQ10 did not show antioxidant activity in cell free system of DPPH radical scavenge, it had a potent antioxidant activity in cell culture system of both silica- and zymosan-induced reactive oxygen species generation using Raw 264.7 cells. This result may be associated with the conversion of CoQ10 to the reduced form (CoQ10H(2)) in the presence of some kinds of intracellular reducing agents. In conclusion, it is considered that CoQ10 appears to have a cutaneous healing effect in vivo, which may be related to the secondary action of CoQ10.

GC1118, an Anti-EGFR Antibody with a Distinct Binding Epitope and Superior Inhibitory Activity against High-Affinity EGFR Ligands.

The EGFR-targeted monoclonal antibodies are a valid therapeutic strategy for patients with metastatic colorectal cancer (mCRC). However, only a small subset of mCRC patients has therapeutic benefits and there are high demands for EGFR therapeutics with a broader patient pool and more potent efficacy. In this study, we report GC1118 exhibiting a different character in terms of binding epitope, affinity, mode of action, and efficacy from other anti-EGFR antibodies. Structural analysis of the EGFR-GC1118 crystal complex revealed that GC1118 recognizes linear, discrete N-terminal epitopes of domain III of EGFR, critical for EGF binding but not overlapping with those of other EGFR-targeted antibodies. GC1118 exhibited superior inhibitory activity against high-affinity EGFR ligands in terms of EGFR binding, triggering EGFR signaling, and proliferation compared with cetuximab and panitumumab. EGFR signaling driven by low-affinity ligands, on the contrary, was well inhibited by all the antibodies tested. GC1118 demonstrated robust antitumor activity in tumor xenografts with elevated expression of high-affinity ligands in vivo, whereas cetuximab did not. Considering the significant role of high-affinity EGFR ligands in modulating tumor microenvironment and inducing resistance to various cancer therapeutics, our study suggests a potential therapeutic advantage of GC1118 in terms of efficacy and a range of benefited patient pool. Mol Cancer Ther; 15(2); 251-63. ©2015 AACR.

Isolation and characterization of a novel myoactive tetradecapeptide-related peptide isolated from the brain of the squid, Todarodes pacificus.

A new bioactive tetradecapeptide, GFKDNVSNRIAHGFamide, was isolated from the brain of the squid, Todarodes pacificus. Using isolated T. pacificus esophagus as a bioassay, the peptide was shown to induce potent contraction of smooth muscle. The threshold concentration for contraction was 5×10(-10) M to 1×10(-9) M. The peptide was homologous to other molluskan (class Gastropoda) and annelid myoactive tetradecapeptides and to some extent, to arthropodan tridecapeptides. A full-length cDNA encoding the biosynthetic precursor of the active peptide was cloned, revealing that the peptide is probably secreted following processing of a prepropeptide containing a signal peptide and prosequences. This is the first myoactive tetradecapeptide (MATP) to be isolated from any mollusk of the class Cephalopoda and we have named it Todarodes tetradecapeptide (TTP).

The analgesic and anti-inflammatory effects of 7-oxosandaracopimaric acid isolated from the roots of Aralia cordata.

The root of Aralia cordata is a traditional medicine for the treatment of inflammation, fever, pain, and spasm in the various diseases in Korea. We isolated a dibenzylbutyrolactone diterpene acid, 7-oxosandaracopimaric acid (OSA), from the ether fraction of Aralia cordata MeOH extract, and studied the effect of OSA on phenylquinone (PQ)-induced writhing syndrome and PQ-induced capillary permeability increase, compound 48/80-induced histamine release by peritoneal mast cells, cycloxygenase (COX) activities, and silica-induced RAW 264.7 cell reactive oxygen species production. OSA (30 mg/kg, p.o.) significantly (p < 0.05) inhibited PQ-induced writhes by 25.8% and the PQ-induced capillary permeability increase levels by 33.13% as compared with PQ control. Furthermore, OSA (10 mM) inhibited COX-1 by 22.82 +/- 1.94%, and COX-2 by 15.86 +/- 1.35%, respectively, to the same extent as indomethacin at the same concentration (10 mM). And OSA (3.0 mM) significantly (p < 0.05) inhibited compound 48/80-induced histamine release from rat mast cells, and its activity was similar to that of celebrex (1 mM), but no piracetam (0.1 mM) inhibited them. OSA did not inhibit ROS production in RAW 264.7 cells. These results indicated that OSA has analgesic and anti-inflammatory effects due to its inhibitory effects on capillary permeability, COX activities, and histamine release.

Antioxidant Effect of CoQ(10) on N-nitrosodiethylamine-induced Oxidative Stress in Mice.

The antioxidant effect of CoQ(10) on N-nitrosodiethylamine (NDEA)-induced oxidative stress was investigated in mice. Food intake and body weight were similar in both CoQ(10) and control groups during the 3-week experimental period. NDEA significantly increased the activities of typical marker enzymes of liver function (AST, ALT and ALP) both in control and CoQ(10) groups. However, the increase of plasma aminotransferase activity was significantly reduced in the CoQ(10) group. Lipid peroxidation in various tissues, such as heart, lung, liver, kidney, spleen and plasma, was significantly increased by NDEA, but this increase was significantly reduced by 100 mg/kg of CoQ(10). Superoxide dismutase activity increased significantly upon NDEA-induced oxidative stress in both the control and CoQ(10) groups with the effect being less in the CoQ(10) group. Catalase activity decreased significantly in both the control and CoQ(10) groups treated with NDEA, again with the effect being less in the CoQ(10) group. The lesser effect on superoxide dismutase and catalase in the NDEA-treated CoQ(10) group is indicative of the protective effect CoQ(10). Thus, CoQ(10) can offer useful protection against NDEA-induced oxidative stress.

The Effect of Caffeic Acid on Wound Healing in Skin-incised Mice.

This study was carried out to investigate the wound healing effect of caffeic acid in skin-incised mice. Caffeic acid showed significant effects on anti-inflammatory activity and wound healing, such as myeloperoxidase activity, lipid peroxidation, phospholipase A(2) activity and collagen-like polymer synthesis, in incised-wound tissue. On the other hand, it significantly stimulated collagen-like polymer synthesis in NIH 3T3 fibroblast cells, while inhibited both silica-induced reactive oxygen species generation and melittin-induced arachidonic acid release and PGE(2) production in Raw 264.7 cells, and histamine release in RBL 2H3 cells stimulated by melittin or arachidonic acid. Therefore, caffeic acid appears to have a potent antioxidant and anti-inflammatory effect in cell culture system, which may be related to wound healing in skin-incised mice.

[Efficacy and safety of long-term lamivudine therapy in the patients with decompensated liver cirrhosis secondary to hepatitis B].

BACKGROUND/AIMS: Lamivudine use in patients with decompensated cirrhosis B has been reported to improve the hepatic function and often delay the need for liver transplantation. In the present study, we evaluated the efficacy and safety of long-term lamivudine therapy in patients with decompensated cirrhosis by comparative study using a matched, untreated cohort. METHODS: 41 patients with decompensated cirrhosis B were included for this study (31 male and 10 female; mean age, 50 years; mean observation period, 18 months). They were divided into two groups: a lamivudine treatment group and an untreated control group. 21 patients in the treatment group were treated with lamivudine 75 or 150 mg daily for at least 12 months. Biochemical and serologic markers were evaluated at two to three-month intervals for all patients. Clinical improvement was defined by a decrease in the Child-Pugh score of at least 2 points. RESULTS: During the observation period, 62% (13/21) was responders, 33% (7/21) was breakthrough, and 5% (1/21) was non-responder in the treated group. The mean Child-Pugh score was significantly improved from 8.6 to 6.0 in the treatment group, but aggravated from 8.7 to 10.0 in the control group during the follow-up. The HBeAg seroconversion rate was 31% in the treatment group (5/16) and none in the control group (0/14). Clinical improvement was observed in fifteen of 21 in the treatment group (71%) and only one of 20 in the control group (5%). According to the treatment responses, clinical improvement was observed in ten of 13 responders (77%), four of 7 breakthrough (57%), and one non-responder. CONCLUSIONS: The long-term administration of lamivudine for patients with decompensated cirrhosis B is effective and safe, although breakthrough and non-response occurred in some patients.