Proteomic analysis of Terminalia chebula extract-dependent changes in human lymphoblastic T cell protein expression.

Terminalia chebula is a native plant from southern Asia to southwestern China that is used in traditional medicine for the treatment of malignant tumors and diabetes. This plant also has antibacterial and immunomodulatory properties. The present study assessed T. chebula extract-dependent protein expression changes in Jurkat cells. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and Ingenuity Pathways Analysis (IPA) were performed to assess protein expression and networks, respectively. A comparative proteomic profile was determined in T. chebula extract (50 µg/mL)-treated and control cells; the expressions of ß-tubulin, ring finger and CHY zinc finger domain containing 1, and insulin-like growth factor 1 receptor kinase were significantly down-regulated in T. chebula extract-treated Jurkat cells. Moreover, the molecular basis for the T. chebula extract-dependent protein expression changes in Jurkat cells was determined by IPA. Treatment with the T. chebula extract significantly inhibited nuclear factor-κB activity and affected the proteomic profile of Jurkat cells. The molecular network signatures and functional proteomics obtained in this study may facilitate the evaluation of potential antitumor therapeutic targets and elucidate the molecular mechanism of T. chebula extract-dependent effects in Jurkat cells.

Terminalia chebula extract acts as a potential NF-κB inhibitor in human lymphoblastic T cells.

Terminalia chebula (TC) is native to southern Asia to southwestern China and is used in traditional medicine for the treatment of human ailments including malignant tumors and diabetes. This plant also has antibacterial and immunomodulatory properties. Nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB) is responsible for the expression of numerous genes involved in cell survival, proliferation, angiogenesis, inflammation, invasion and metastasis, among other processes. This study aims to assess the NF-κB inhibitory effect of TC extract in human lymphoblastic T (Jurkat) cells. The effects of TC extract were investigated using the FRET-based Gene Blazer technique in transfected Jurkat-NF-κB-RE-bla cells. The concentration of TC extract required for NF-κB inhibition was determined by a cell proliferation assay. Treatment with TC extract (50 µg/mL) inhibited NF-κB activity and protected against IκBα degradation and strongly suppressed IκBα phosphorylation in Jurkat-NF-κB-RE-bla cells. This treatment might be crucial for inhibiting NF-κB translocation and activation. In addition, the TC extract downregulated certain NF-κB regulated genes, including IL-8 and MCP-1, in Jurkat-NF-κB-RE-bla cells. Moreover, gallic acid was identified from the TC extract demonstrating its ability to inhibit NF-κB activity in Jurkat-NF-κB-RE-bla cells. Further studies to identify the role of gallic acid in NF-κB inhibition may uncover the crucial antiinflammatory and antitumor properties of the TC extract.

Melanogenesis inhibitory effects of methanolic extracts of Umbilicaria esculenta and Usnea longissima.

The primary objective of this study was to assess the in vitro melanogenesis inhibitory effects of methanolic extracts of the edible and medicinal lichens, Umbilicaria (Gyrophora) esculenta and Usnea longissima. The quantities of the total phenolic compounds of methanolic extract of the two lichen extracts were determined to be 1.46% and 2.62%, respectively. In order to evaluate the antioxidative effects of the extracts, we also measured electron donating abilities (EDA) and lipid peroxidation rates. The EDA values measured by the reduction of 1.1''-diphenyl-2-picrylhydrazyl (DPPH) were 72.8% and 80.7% for the extracts, with SC50 (median scavenging concentration) values of 1.29+/-0.05 mg/ml and 1.03+/-0.06 mg/ml, respectively. The rates of inhibition of lipid peroxidation using linoleic acid were 92.1% and 97.3% for the extracts, with IC50 (median inhibitory concentration) values of 0.57+/-0.05 mg/ml and 0.53+/-0.06 mg/ml, respectively. The inhibitory rates of the extracts against tyrosinase were 67.4% and 84.8%, respectively. The extracts were shown to reduce melanin formation in human melanoma cells. Melanin contents in the samples treated with 0.01% and 0.1% U. esculenta were 47.1% and 31.2%, respectively, and those treated with 0.01% and 0.1% Usnea longissima were 51.1% and 34.9%, respectively, whereas a value of 54.0% was registered when ascorbic acid was utilized as a positive control. In addition to direct tyrosinase inhibition, it was determined that the lichen extracts affected the activity of tyrosinase via the inhibition of tyrosinase glycosylation. As a result, the methanolic extracts of U. esculenta and Usnea longissima evidenced melanogenesis inhibitory effects, which occurred via multiple routes.

Antithrombotic activity of methanolic extract of Umbilicaria esculenta.

Antithrombotic activity of methanolic extract of an edible lichen, Umbilicaria esculenta, was evaluated on platelet aggregation in vitro and pulmonary thrombosis in vivo. The extract showed concentration dependent inhibitory effects on platelet aggregation induced by ADP, with IC(50) value of 2.4 mg/mL. Orally administered extract protected mice against thrombotic death or paralysis induced by collagen and epinephrine in a dose dependent manner. It produced a significant inhibition of thrombotic death or paralysis at over 100 mg/kg body weight, while aspirin produced a significant inhibition of thrombosis at 10-20 mg/kg body weight. Mouse tail bleeding time was significantly prolonged by addition of the extract. On the other hand, the extract did not show any fibrinolytic activity and alter coagulation parameters such as activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) in rat platelet. These results suggested that the antithrombotic activity of Umbilicaria esculenta extract might be due to antiplatelet activity rather than anticoagulation activity.

Antiplatelet and antithrombotic activities of methanol extract of Usnea longissima.

The antiplatelet and antithrombotic activities of a methanol extract of a medicinal lichen, Usnea longissima, were investigated on platelet aggregation in vitro and on pulmonary thrombosis in vivo. The extract showed concentration dependent inhibitory effects on ADP-induced platelet aggregation, with an IC50 value of 3.6 mg/mL. Using an in vivo mouse thrombotic model in which mice were challenged with an intravenous injection of collagen and epinephrine mixture, oral administration of the extract prior to the injection produced a significant inhibition of thrombotic death or paralysis at 100-200 mg/kg body weight. Aspirin, a representative antiplatelet drug, produced a significant inhibition of thrombotic death at 10-20 mg/kg body weight. The mouse tail bleeding time was significantly prolonged by the addition of the extract. On the other hand, the extract did not show any fibrinolytic activity or alter the coagulation parameters such as activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) in rat platelets in vitro. These results suggested that the antithrombotic activity of U. longissima extract might be due to antiplatelet activity rather than anticoagulant activity.