In vitro immunomodulatory activity of flavonoid glycosides from Urtica dioica L

The major compounds isolated from the methanolic extract of the aerial parts of Urtica dioica L. were determined as quercetin-3-O-rutinoside (1). kaempherol-3-O-rutinoside (2). and isorhamnetin-3-O-glucoside (3). by chromatographic, chemical (acidic hydrolysis) and spectral (UV, IR, (1)H-NMR, (13)C-NMR) methods. Their immunomodulatory activities were studied in vitro by chemotaxis (Boyden Migration Chamber) and intracellular killing activity (NBT reduction) tests. Compounds 1, 2, 3 and the total flavonoid fraction were determined to have significant chemotactic effects in 4, 8, 16 microg doses. According to the results of the NBT reduction test, all flavonoid glycosides showed high intracellular killing activity. The results of both assays confirmed the immunostimulatory activity of the flavonoid fraction and the isolated flavonoid glycosides on neutrophils suggesting that they could possibly be useful for treating patients suffering from neutrophil function deficiency and chronic granulomatous diseases.     

Chemical constituents and antimicrobial activity of medicinal plants from Ghana: Cassia sieberiana, Haematostaphis barteri, Mitragyna inermis and Pseudocedrela kotschyi

The antimicrobial activity of the sequential n-hexane, acetone and 50% aqueous methanol extracts of leaves, stem bark and roots of four species of medicinal plants, Cassia sieberiana DC. (Leguminosae), Haematostaphis barteri Hook. f. (Anacardiaceae), Mitragyna inermis (Willd.) O. Kuntze (Rubiaceae) and Pseudocedrela kotschyi (Schweinf.) Harms (Meliaceae), from Ghana were tested against Bacillus subtilis, Pseudomonas syringae and Cladosporium herbarum using TLC direct-autobiographic methods. Extracts from leaves, stem bark and roots of the four species gave a positive result against at least one test organism. Twelve of the 36 extracts were active against B. subtilis, four extracts were active against P. syringae and six were active against C. herbarum. Preliminary chemical analysis revealed the presence of flavonoids, stilbenes and alkaloids. This is the first report of a stilbene from the Anacardiaceae.     

Inhibition of Osteoclast Differentiation by Ginsenoside Rg3 in RAW264.7 Cells via RANKL,JNK and p38 MAPK Pathways Through a Modulation of Cathepsin K: An In Silico and In Vitro Study

Various studies have demonstrated that overexpression of cathepsin K (Cat‐K) causes excessive bone loss, which ultimately leads to a variety of bone diseases including osteoporosis. Therefore, inhibition of Cat‐K signifies a potential therapeutic target in osteoporosis treatment. Ginsenoside Rg3 is one of the most promising compound of Panax ginseng Meyer (P. ginseng) with numerous biological activities. Thus, in recent study the inhibitory effect of Rg3 isolated from P. ginseng was investigated in order to impede the osteoclast activity by an in silico approach followed by in vitro study validation using RAW264.7 cells through the investigation of different biological activity prediction such as absorption distribution metabolism and excretion (ADMET) properties against Cat‐K protein. The docking results of our study showed that Rg3 is a non‐toxic compound and may act as a drug‐like molecule. Additionally, the molecular interaction of Rg3 with the active residues of Cat‐K markedly describes its inhibitory effects on osteoclastogenesis. Findings of the present study exhibited that Rg3 significantly reduced receptor activator of nuclear factor kappa B ligand (RANKL)‐induced tartrate‐resistant acid phosphatase (TRAP) activity, pit formation (actin rings), and TRAP‐positive multinucleated cells development in RAW264.7 cells. Furthermore, Rg3 dose‐dependently reduced the mRNA expression levels of osteoclast‐specific markers such as RANK, TRAP, and Cat‐K induced by RANKL through the down regulation of p38, extracellular signal‐regulated kinase, and c‐Jun N‐terminal kinase (JNK) pathways. In conclusion, in silico docking study and in vitro validation together suggested that Rg3 inhibits osteoclastogenesis and reduces bone resorption through the inhibition of Cat‐K. Therefore, Rg3 might be a useful therapeutic agent for the treatment of osteoporosis and proper bone formation. Copyright © 2015 John Wiley & Sons, Ltd.     

Therapeutic Effect of Rhizoma Dioscoreae Nipponicae on Gouty Arthritis Based on the SDF‐1/CXCR 4 and p38 MAPK Pathway: An In Vivo and In Vitro Study

Rhizoma Dioscoreae Nipponicae (RDN) is a widely used traditional Chinese herb, which is used to treat arthroncus, arthrodynia and arthritis. As is known to us, inflammatory mechanisms have played an important role in the occurrence, course and prognosis of gouty arthritis (GA). The aim of this study was to determine the characteristic expressed proteins of synovium in GA rat and synovial cell. The rat model of GA was induced by monosodium urate (MSU) crystal. Tissue samples were assayed by immunohistochemical method. The effects of RDN on Stromal cell‐derived factor 1 (SDF‐1), CXCR 4 and p38 mitogen‐activated protein kinase (MAPK) were investigated in MSU crystal‐induced rat. The levels of SDF‐1 and mitogen‐activated kinase kinase (MKK) 3/6 were measured by Western Blot in interleukin‐1β (IL‐1β) incubated fibroblast‐like synoviocytes (FLS). A significant increase in the levels of SDF‐1, CXCR 4 and p38 MAPK were observed in MSU crystal‐induced rat. The increased SDF‐1 and MKK 3/6 levels were observed in IL‐1β incubated FLS. With the treatment of RDN, the above changes were reverted back to near normal levels. RDN might have some therapeutic effects on GA through SDF‐1/CXCR 4 and p38 MAPK pathway, and dioscin may be the active compound in RDN to exert therapeutic effect on GA. Copyright © 2013 John Wiley & Sons, Ltd.     

Labda‐8(17),12,14‐trien‐19‐oic Acid Contained in Fruits of Cupressus sempervirens Suppresses Benign Prostatic Hyperplasia in Rat and In Vitro Human Models Through Inhibition of Androgen and STAT‐3 Signaling

Fruit extract of Cupressus sempervirens (CS), which is used traditionally to treat Benign Prostatic Hyperplasia (BPH)‐like urinary symptoms in patients, was scientifically validated for anti‐BPH activity. The ethanolic fruit extract of CS inhibited proliferation of human BPH‐stromal cells and the activity was localized to its chloroform‐soluble, diterpene‐rich fraction. Eight major diterpenes isolated from this fraction exhibited moderate to potent activity and the most active diterpene (labda‐8(17),12,14‐trien‐19‐oic acid) exhibited an IC₅₀ of 37.5 μM (antiproliferative activity against human BPH‐stromal cells). It significantly inhibited activation (phosphorylation) of Stat‐3 in BPH‐stromal cells and prevented transactivation of androgen sensitive KLK3/PSA and TMPRSS2 genes in LNCaP cells. Labda‐8(17),12,14‐trien‐19‐oic acid‐rich CS fraction prevented prostatic hyperplasia in rat model and caused TUNEL labeling of stromal cells with lower expressions of IGF‐I, TGF‐ß and PCNA, and bcl‐2/bax ratio. Human BPH tissues exhibited precise lowering of stromal component after incubation in labda‐8(17),12,14‐trien‐19‐oic acid, ex vivo. We conclude that labda‐8(17),12,14‐trien‐19‐oic acid contained in CS exhibits anti‐BPH activity through inhibition of stromal proliferation and suppression of androgen action in the prostate, presenting a unique lead structure for further optimization of anti‐BPH activity. Copyright © 2014 John Wiley & Sons, Ltd.