Structural characterization of anti-UV components from Sasa senanensis Rehder extract.

BACKGROUND: Previous studies have shown antiviral, antibacterial and anti-inflammatory activity of alkaline extract of the leaves of Sasa senanensis Rehder (SE). However, active components have not been identified. We isolated the substances that exhibit anti-UV activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity from SE and estimated their putative structures. MATERIALS AND METHODS: The anti-UV substances (SEE-1 and SEE-2) were isolated from SE by ethanolic extraction, Wakosil chromatography and recycled high-performance liquid chromatography (HPLC) at a yield of 0.22 and 0.18%, respectively. The structural analysis was carried out with (1)H-nuclear magnetic resonance (NMR), (13)C-NMR and UV absorption. RESULTS: SEE-1 exhibited approximately four-fold higher anti-UV activity and slightly lower DPPH radical-scavenging activity, compared to SE. SEE-1 was identified as p-coumaric acid derivative(s), a lignin precursor. CONCLUSION: The present study demonstrated for the first time the presence of lignin precursors in SE, which may explain why SE exhibits many of the properties of lignin-carbohydrate complexes.

Pilot clinical study of Sasa senanensis Rehder leaf extract treatment on lichenoid dysplasia.

BACKGROUND: Previous studies have shown antiviral, antibacterial, and anti-inflammatory activity of alkaline extract of the leaves of Sasa senanensis Rehder (SE). Here, we investigated whether SE is effective on oral lichenoid dysplasia and osteoclastogenesis. MATERIALS AND METHODS: A male patient with white lacy streaks in the oral mucosa was orally administered SE three times a day for 11 months. The area of white streaks was monitored by intraoral photography. Interleukin-6 and -8 in the saliva were determined by enzyme-linked immunosorbent assay. Osteoclastogenesis of mouse macrophage-like RAW264.7 cells, induced by receptor activator of nuclear factor-κB ligand (RANKL) was monitored by tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cell formation. RESULTS: Long-term treatment with SE progressively reduced both the area of white steaks and the levels of salivary interleukin-6 and -8. SE significantly inhibited the macrophage differentiation towards osteoclasts. CONCLUSION: The present study suggests the therapeutic potential of SE towards oral diseases.

Anti-UV/HIV activity of Kampo medicines and constituent plant extracts.

AIM: In order to search for new biological activities of Kampo medicines and their constituent plant extracts, we investigated whether they protect the cells from the cytotoxicity induced by UV irradiation and human immunodeficiency virus (HIV) infection. MATERIALS AND METHODS: Anti-UV/HIV activity (SI value) was evaluated as the ratio of the CC(50) (concentration that reduced the viable cell number by 50%) to the EC(50) (the concentration that increased the viability of UV-irradiated or HIV-infected cells to 50%): SI=CC(50)/EC(50). The content of glycyrrhizin in each sample was determined by high performance liquid chromatography (HPLC). Caspase-3/-7 activity was assayed by cleavage of poly ADP ribose polymerase using western blot analysis. RESULTS: Among 25 plant extracts, Gardenia fruit had the highest anti-UV activity (SI≥8.0), followed by Glycyrrhiza (SI=4.3), Coptis rhizoma (SI=1.5), Cimicifuga rhizoma (SI>1.4), Saposhnikovia root (SI>1.3) and Japanese Gentian (SI>1.1). Among ten Kampo medicines, Unseiin and Hangesyashinto (SI>4.9) had the highest anti-UV activity, followed by Shosaikoto (SI>4.3), Saireito (SI>3.4), Rikkosan (SI>1.2) and Kikyoto (SI=1.1). Glycyrrhiza inhibited UV-induced caspase-3/-7 activation. Only Polyporus sclerotium (SI>4.4), Gardenia fruit (SI>2.7), Atractylodes lancea rhizoma (SI>1.9), Cnidium rhizoma (SI>1.5) and Japanese Angelica root (SI>1.1) exhibited some anti-HIV activity. There was no apparent correlation of their anti-UV/HIV activity and content of glycyrrhizin, a major component of Glycyrrhiza, which exhibited much higher anti-UV activity (SI=20.6) and some anti-HIV activity (SI>2.0). CONCLUSION: The present study suggests the involvement of substances other than glycyrrhizin in the anti-UV/HIV activity of Kampo medicines and their constituent plant extracts.

Biological activity of SE-10, granulated powder of Sasa senanensis Rehder leaf extract.

BACKGROUND: We have previously reported that alkaline extract of Sasa senanensis leaves (SE) showed potent anti-HIV, anti-UV and radical scavenging activity. In the present study, we investigated the biological activities of SE-10, a granulated powder of SE supplemented with lactose, lactitol, trehalose and tea extract. MATERIALS AND METHODS: Cell viability of mock-infected, HIV-infected, and UV-irradiated cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Scavenging activity of superoxide anion and hydroxyl radicals was determined by electron-spin resonance spectroscopy. Cytochrome P-450 (CYP)3A4 activity was measured by ß-hydroxylation of testosterone in human recombinant CYP3A4. RESULTS: SE-10 had slightly higher anti-HIV and anti-UV activities, but slightly lower radical-scavenging and CYP3A4-inhibitory activities, as compared with SE. CONCLUSION: The present study demonstrates that the biological activities of SE were well preserved during the manufacturing process of SE-10.

Comparative study of biological activity of three commercial products of Sasa senanensis Rehder leaf extract.

BACKGROUND: We have previously reported that alkaline extract of Sasa senanensis leaves (SE) has several biological activities characteristic of lignin-carbohydrate complex (LCC). In the present study, we compared the biological activity of three commercially available products of SE (products A, B and C). MATERIALS AND METHODS: Cell viability of mock-infected, HIV-infected, UV-irradiated cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Radical intensity was determined by electron spin resonance spectroscopy. Cytochrome P-450 (CYP)3A4 activity was measured by ß-hydroxylation of testosterone in human recombinant CYP3A4. RESULTS: Product A is a pure SE that contains Fe(II)-chlorophyllin, whereas products B and C contain Cu(II)-chlorophyllin and less LCC. Product C is supplemented with ginseng and pine (Pinus densiflora) leaf extracts. Product A exhibited 5-fold higher anti-HIV, 4-fold higher anti-UV, 5-fold higher hydroxyl radical-scavenging, and 3-fold lower CYP3A4 inhibitory activities as compared to those of product B, and 5-fold higher, 1.5-fold higher, comparable, and 7-fold lower activities, respectively, as compared to those of product C. CONCLUSION: The present study demonstrates for the first time the superiority of product A over products B and C, suggesting the beneficial role of LCC and Fe(II)-chlorophyllin.

Anti-UV activity of Lentinus edodes mycelia extract (LEM).

BACKGROUND: Using our recently established simple method for evaluating protective activity from ultraviolet ray injury (referred to as 'anti-UV activity'), the effectiveness of various antioxidants and plant extracts was investigated. MATERIALS AND METHODS: HSC-2 human oral squamous cell carcinoma cells were exposed to UV irradiation (wavelength: 253.7 nm, 6 J/m²) in phosphate-buffered saline (PBS(-)) containing various concentrations of samples and then incubated for 48 hours in regular culture medium to determine the viable cell number by the MTT method. RESULTS: Among the representative antioxidants, sodium ascorbate showed the most potent anti-UV activity, whereas catalase and N-acetyl-L-cysteine were inactive. Lentinus edodes mycelia extract (LEM) showed comparable anti-UV activity to sodium ascorbate. Hot water extracts of green tea and coffee, and PET-bottled of green tea extract showed slightly less, but noticeable anti-UV activity. On the other hand, hot water extracts of black tea and Jasmine tea, and PET-bottled of oolong tea, barley tea and Kohki tea were inactive. LEM was separated by gel filtration chromatography into four fractions from high to low molecular weight: polysaccharide, large and small lignin-carbohydrate complexes, and sugars. Anti-UV activity was shown by the lignin-carbohydrate fractions, but not the polysaccharide and sugar fractions. LEM, at high concentration, slightly enhanced the anti-UV activity of sodium ascorbate. CONCLUSION: LEM may be applicable as a UV-protective agent.

Anti-UV Activity of Alkaline Extract of the Leaves of Sasa senanensis Rehder.

BACKGROUND: We have previously reported several lignin-like activities of the alkaline extract of the leaves of Sasa senanensis Rehder (SE), such as anti-HIV and radical scavenging activity. As an extension, possible SE protection of cells from ultraviolet (UV)-induced injury (referred to "anti-UV activity") was investigated. MATERIALS AND METHODS: HSC-2, human oral squamous cell carcinoma cells, were exposed to UV irradiation in phosphate-buffered saline containing SE or its fractions, and then incubated for 48 hours in fresh regular culture medium to determine the viable cell number by the MTT method. SE was separated by gel filtration chromatography into the following four fractions: polysaccharide, large and small lignin-carbohydrate complexes and lower molecular weight polyphenol fractions (Fr. I). RESULTS: The anti-UV activity of SE was slightly less than that of sodium ascorbate, but higher than that of gallic acid, (-)-epigallocatechin gallate, chlorophyll a and chlorophyllin. The combination of SE and sodium ascorbate gave synergistic anti-UV activity. On gel filtration fractionation, the majority of the anti-UV activity was recovered from Fr. I, which was eluted as a single peak on high performance liquid chromatography (HPLC). CONCLUSION: The anti-UV activity of SE further suggests its potential as an alternative medicine.

Biological activity of luteolin glycosides and tricin from Sasa senanensis Rehder.

BACKGROUND: In contrast to the several reports of alkaline extracts (Sasa-health, SE), no study of flavonoids from the leaves of S. senanensis has been reported. Four flavonoids were isolated from this plant species and their biological activities were investigated. MATERIALS AND METHODS: Luteolin 6-C-ß-D-glucoside [1], luteolin 7-O-ß-D-glucoside [2], luteolin 6-C-α-L-arabinoside [3] and tricin [4] were extracted from the leaf of S. senanensis with methanol, partitioned with ethyl acetate, separated by Sephadex LH-20 and purified by high-performance liquid chromatography (HPLC). The structure was determined by ultraviolet (UV) spectra, high-resolution mass spectra (HR-MS) and nuclear magnetic resonance (NMR). RESULTS: The luteolin glycosides, 1-3 showed no cytotoxicity against the human normal oral cells and oral squamous cell carcinoma cell lines used up to 0.8 mg/ml, whereas 4 was highly cytotoxic. The luteolin glycosides 1-3 protected the cells from UV induced cytotoxicity, more efficiently than 4. The anti-HIV activity of 4 (Selectivity index, SI=27) was much higher than that of the luteolin glycosides (SI=2-7), but lower than that of SE (SI=40). The scavenging activity of 1-3 against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radicals was comparable with that of quercetin and, much higher than that of 4. CONCLUSION: The luteolin glycosides from S.senanensis show several new biological properties distinct from tricin and the anti-UV activity of the luteolin glycosides may be derived from their radical scavenging activity.

Quest for anti-inflammatory substances using IL-1ß-stimulated gingival fibroblasts.

BACKGROUND: We have previously reported that azulene-related compounds, and alkaline extract of Sasa senanensis Rehder potently inhibited nitric oxide (NO) production by lipopolysaccharide (LPS)-stimulated mouse macrophages. We investigated here whether they can inhibit pro-inflammatory cytokine production, by activated human gingival fibroblast (HGF). MATERIALS AND METHODS: HGF was established from the periodontal tissues of extracted tooth. Viable cell number was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Production of Prostaglandin E(2) (PGE(2)) and cytokines was determined by enzyme immunoassay, and enzyme-linked immunosorbent assay, respectively. RESULTS: Interleukin (IL)-1ß did not inhibit, but rather slightly stimulated the growth of HGF cells. IL-1ß stimulated the production of PGE(2), IL-6, IL-8 and monocyte chemotactic protein-1 very potently, but not that of nitric oxide and tumor necrosis factor-α. Native LPS and synthetic lipid A from E. coli and P. gingivalis was much less stimulatory. Dexamethasone, not indomethacin, was an efficient inhibitor of IL-8 production. Among five azulene-related compounds, benzo[b]cyclohepta[e][1,4]thiazine most potently inhibited the IL-8 production by HGF cells, as well as NO production by activated RAW264.7 cells. The alkaline extract of Sasa senanensis Rehder significantly inhibited IL-8 production, without affecting the cell viability. CONCLUSION: The present system may be applicable for use in the search for anti-gingivitis substances.

Botanical origin of mei-gui hua (petal of a Rosa species).

Mei-gui hua has been used as a crude drug in traditional medicine and as herbal tea in China. The scientific name of Mei-gui is Rosa rugosa thunb. However, the morphological characteristics and botanical ecology of Mei-gui were different from those of R. rugosa. Since the botanical origins of Mei-gui cultivated in China have not yet been clarified, we compared Mei-gui and R. rugosa in terms of their morphological characteristics, phylogenetic analysis, and phytochemical studies. Our research suggested that Mei-gui cultivated around Tarim Basin in Xinjiang Province showed homology to Rosa gallica, while those cultivated in the northeastern parts of China are considered to be hybrids of R. rugosa.