Quantitation and Radical Scavenging Activity Evaluation of Iridoids and Phenylethanoids from the Roots of Phlomis umbrosa (Turcz.) using DPPH Free Radical and DPPH-HPLC Methods, and their Cytotoxicity

The roots of Phlomis umbrosa (Turcz.) (Phlomidis Radix) have been traditionally used to treat cold, reduce swelling and staunch bleeding. Four iridoids (1 – 3 and 5) and six phenylethanoid derivatives (4, and 6 – 10) were isolated from the roots of P. umbrosa. A simple, sensitive, and reliable analytical HPLC/PDA method was developed, validated, and applied to determine 10 marker compounds in Phlomidis Radix. Furthermore, the isolates were evaluated for cytotoxic and anti-oxidant activities as well as DPPH-HPLC method. Among them, compounds 4 and 6 – 9 displayed potent anti-oxidant capacities using DPPH assay with IC50 values of 27.7 ± 2.4, 10.2 ± 1.1, 18.0 ± 0.8, 19.1 ± 0.3, and 19.9 ± 0.6 µM, and compounds 6, 8, and 9 displayed significant cytotoxic activity against HL-60 with IC50 values of 35.4 ± 3.1, 18.6 ± 2.0, and 42.9 ± 3.0 µM, respectively.

A new isobenzofuranone derivative from Phlomis betonicoides and its antibacterial activity / 中国中药杂志

A new isobenzofuranone derivative has been isolated from Phlomis betonicoides by using various chromatographic techniques, including silica gel, Sephadex LH-20, MCI-gel resin and RP-HPLC. This compound was determined as 5-(3-hydroxypropyl)-2,2-dimethyl-2-furo[3,4-]chromen-7(9)-one (1) by NMR, MS, IR and UV spectroscopic data. Compound 1 showed potent antibacterial activity with an MIC₉₀ value of (58.4 ± 4.2) mg·L⁻¹ for methicillin resistant Staphylococcus aureus (MRSA) strain [levofloxacin as a control with MIC₉₀ value of (52.8±4.6) mg·L⁻¹].

Antioxidant Compounds Isolated from the Roots of Phlomis umbrosa Turcz

Two triterpenoids, arjunolic acid (1), belleric acid (2), five phenylethanoids, martynoside (3), orobanchoside (4), 3,4-dihydroxyphenethylalcohol-6-O-caffeoyl-β-D-glucoside (5), leucosceptoside B (6), lunariifolioside (7), four phenolic acids, ferulic acid (8), syringic acid (9), vanillic acid (10), 4-hydroxybenzoic acid (11), and one lignan, (+)-syringaresinol-β-D-glucoside (12), were isolated from the roots of P. umbrosa. All isolated compounds were explored for their antioxidant potential in the DPPH and ABTS assays. In DPPH assay, compound 5 showed high antioxidant capacity. Compounds 3, 4, 6, and 7 displayed considerable antioxidant activities. In addition, compounds 5–7 exhibited potential antioxidant capacities in the ABTS assay.

GLC-MS profiling of non-polar extracts from Phlomis bucharica and P. salicifolia and their cytotoxicity / Perfiles GLC-MS de extractos no polares de Phlomis bucharica y P. salicifolia y su citotoxicidad

Phlomis species (Phlomis bucharica Regel and P. salicifolia Regel) have been traditionally used by Uzbek people as stimulant, tonic, diuretic, and in the treatment of ulcers, hemorrhoids, wounds and gynecological problems. In the present study, we characterized the chemical composition of non-polar extracts from P. bucharica and P. salicifolia by high resolution GLC-MS and evaluated their cytotoxicity. Concentrations of hexadecanoic acid in hexane and chloroform extracts were higher in P. bucharica than in P. salicifolia. 1,8- Cineol, camphor, borneol, alfa-terpinol, thymol, and isobornyl acetate were detected in P. bucharica but not in P. salicifolia. About 45 components were identified in P. bucharica and 40 in P. salicifolia. The chloroform extract from P. bucharica showed cytotoxicity in HeLa and HL-60 cells, with IC50 values of 26.07 and 29.42 ug/ml, respectively.

Las especies Phlomis (Phlomis bucharica Regel y P. salicifolia Regel) se han utilizado tradicionalmente por la gente de Uzbekistán como estimulante, tónico, diurético, y en el tratamiento de las úlceras, hemorroides, heridas y problemas ginecológicos. En el presente estudio, hemos caracterizado la composición química de los extractos no polares de P. bucharica y P. salicifolia por GLC-MS de alta resolución y se evaluó su citotoxicidad. Las concentraciones de ácido hexadecanoico en extractos de hexano y cloroformo fueron mayores en P. bucharica que en P. salicifolia. 1,8-cineol, alcanfor, borneol, se detectaron alfa-terpinol, timol, y acetato de isobornilo en P. bucharica pero no en P. salicifolia. Cerca de 45 componentes fueron identificados en P. bucharica y 40 en P. salicifolia. El extracto de cloroformo a partir de P. bucharica mostró citotoxicidad en células HL-60 y HeLa, con valores de CI 50 de 26,07 y 29,42 ug/ml, respectivamente.

Isolation, idetification and anti-HIV-1 integrase activity of culturable endophytic fungi from Tibetan medicinal plant Phlomis younghusbandii Mukerjee / 药学学报

A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.

The Effects of Phlomis umbrosa Turcz on Osteoclast Differentiation / 체질인류학회지

Many bone-related diseases such as osteoporosis, rheumatoid arthritis are occurred by excessive bone resorbing activity of osteoclasts. Recently, many studies have been proceeded to find out the new therapeutic materials from natural products of plants. Phlomis umbrosa Turcz, one of the natural products of plants has been known to improve bone health. However, the precise effects and treatment mechanisms of Phlomis umbrosa Turcz about bone diseases has been unknown. So, we examined the effects of Phlomis umbrosa Turcz on expression of receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) and bone resorption. Also, we investigated the treatment mechanisms of Phlomis umbrosa Turcz relating to osteoclast differentiation. Here, we showed that Phlomis umbrosa Turcz significantly suppressed RANKL-induced osteoclast differentiation and bone resportion. Furthermore, Phlomis umbrosa Turcz suppressed the activation of NF-kappaB in bone marrow macrophage treated RANKL and M-CSF. The mRNA expression of c-Fos, nuclear factor of activated T-cells (NFAT)c1, osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphatase (TRAP) in BMMs was inhibited by Phlomis umbrosa Turcz. Integrin alphanu, beta3 relating to cell adhesion and dendritic cell-specific transmembrane protein (DC-STAMP) relating to the structure of filamentous actin (F-actin) ring and cathepsin K relating to bone resorbing activity are disrupted too. These results suggest that Phlomis umbrosa Turcz will be a good materials to treat bone diseases like osteoporosis.

Determination of five iridoid glycosides in Phlomis younghusbandii by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an RP-HPLC method for the determination of five iridoid glycosides in Phlomis younghusbandii.</p><p><b>METHOD</b>HPLC analysis was performed on a Symmetry C18 (4.6 mm x 150 mm, 5 microm, Waters) column eluted with acetonitrile (A) and water (B) in gradient elution. The gradient program was as follows: 0-5 min kept 7% A; 5-10 min changed to 12% A; 10-40 min kept 12% A. The flow rate was 1.0 mL x min(-1). The column temperature was 20 degrees and the detection wavelength was 235 nm.</p><p><b>RESULT</b>The linear ranges of sesamoside, shanzhiside methyl ester, 7, 8-dehydropenstemoside, penstemoside and 8-O-acetylshanzhiside methyl ester were 0.050-0.650 (r = 0.999 3), 0.050-0.350 (r = 0.999 5), 0.040-0.280 (r = 0.999 4), 0.010-0.070 (r = 0.999 6), 0.040-0. 280 (r = 0.999 7) g x L(-1), respectively. The average recoveries (n = 6) of them were all between 96% and 104%, RSD < 5.0%.</p><p><b>CONCLUSION</b>The method is simple, accurate, repeatable and stable, which can be used for quality control of P. younghusbandii.</p>

Antitumor effect of phlomio1 in vivo and in vitro / 中国中药杂志

To study the antitumor effect of phlomiol extracted from Phlomis younghusbandii in vivo and in vitro. The inhibitory effect of phlomiol on two kinds of human tumor cells proliferation was assayed by MTT method. Transplant tumor models of S180 and H22 were used. After transplantation, different doses of phlomiol were given to the mice for 14 days. The inhibitory rates were calculated. MTT method was used to assess the proliferation of T spleen lymphocyte cells and the activity of NK cells in tumor-bearing mice with S180. Phlomiol (50-100 mg x L(-1)) inhibited the proliferation of three kinds of tumor cells in vitro, antitumor effect of phlomiol was in a dose-dependent manner (r = 0.989, P < 0.05). The inhibitory rates of phlomiol (2.5, 5, 10 mg x kg(-1)) were 28.5%-65.0% and 35.0%-74.5% in tumor-bearing mice with S180 and H22 respectively, It could stimulate the spleen T-cells in tumor-bearing mice with S180 and increase the activity of the NK cells. Phlomiol could inhibit the proliferation of three kinds of tumor cells in vitro, present antitumor effect on the tumor-bearing mice, and improve the immunological function.

Estimation of phylogenetic relationships of Phlomis species based on seed protein polymorphism

In this study, phylogenetic relationships among 39 Phlomis taxa were investigated based on seed protein profiles produced by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). A total of 21 polypeptide bands were scored, of which, 19 were polymorphic among the taxa of the genus Phlomis. A distance matrix was generated from the similarity matrix which was computed by using Jaccard’s similarity coefficients, based on polymorphic bands and then an UPGMA tree was established through cluster analysis performed on the distance matrix. Genetic distances ranged from 0.00 to 0.50 within subsection Dendrophlomis; from 0.00 to 0.625 within subsection Gymnophlomis and from 0.00 to 0.769 within subsection Oxyphlomis. The UPGMA tree formed four groups. The topology of the tree is in agreement with the taxonomic view regarding the section Phlomis as it is divided into three subsections as Dendrophlomis, Gymnophlomis and Oxyphlomis based on morphological characters. The grouping pattern of the tree also indicated that subsection Dendrophlomis is more closely related to subsection Gymnophlomis than subsection Oxyphlomis.

A new furanolabdane diterpene glycoside from Phlomis younghusbandii Mukerjee / 药学学报

In order to find the compound basis of Phlomis younghusbandii Mukerjee that related to pharmacodynamic action, various chromatographic techniques were used to separate and purify the constituents of this plant, and physicochemical and spectral data were used to identify the structures of obtained compounds. A new furanolabdane diterpene glycoside, named as phlomisoside F, was isolated and identified, which was 15,16-epoxy-8(9),13(16), 14-labdatrien-7-ketone-19-oic acid-beta-D-glucopyranosyl ester.

Chemical constituents from roots of Phlomis umbrosa / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Phlomis umbrosa.</p><p><b>METHOD</b>Chemical constituents were isolated by repeated column chromatography (Toyopearl HW-40C and preparative HPLC). The structures were elucidated on the basis of spectral data analysis.</p><p><b>RESULT</b>Thirteen compounds were identified as oleanolic acid (1), corosolic acid (2), hederagenin (3), arjunolic acid (4), belleric acid (5), 3beta-hydroxy-29-al-12-en-28-oleanoic acid (6), butyrospermol (7), beta-sitosterol (8), daucosterol (9), caffeic acid (10), vanillic acid (11), p-hydroxy-benzoic acid (12), 3, 4-dihydroxy-benzoic acid (13).</p><p><b>CONCLUSION</b>Compounds 3 9, 11-13 were isolated from the plant for the first time.</p>

Two iridoid glycoside from roots of Phlomis medicinalis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the roots of Phlomis medicinalis.</p><p><b>METHOD</b>The compounds were isolated and repeatedly purified by macroporous resin, silica gel column chromatography, TLC and PREP-HPLC. Their structures were elucidated by physical and chemical properties and NMR spectra.</p><p><b>RESULT</b>Two iridoid glucosides were obtained and elucidated as 7-epilamalbide (1), chlorotuberoside (2).</p><p><b>CONCLUSION</b>Compound 1 was a new compound, compound 2 was isolated from the plant for the first time.</p>

Phylogenetic relationships among species of the subsection Dendrophlomis Bentham

This study used randomly amplified polymorphic DNA markers to determine genetic relationships among species of the subsection Dendrophlomis. Twenty accessions of the eleven Phlomis taxa were evaluated to determine genetic variability using fourteen ten mer primers selected from a 125 random oligonucleotide set. These 14 selected primers generated 85 RAPD bands that ranged in size from 200 to 1200 base pairs. Of the total bands, 88 percent (75) were polymorphic among the samples. Genetic distances among accessions were computed to produce a dendrogram based on UPGMA. Genetic distances ranged from 0.133 (between P. amanica and P. monocephala) to 0.494 (between P. chimerae and P. lunariifolia). The UPGMA tree based on distances has two major groups. The first comprised 9 taxa that were clustered into two subgroups. The first subgroup consisted of P. viscosa, P. lycia, P. amanica and P. monocephala while the second comprised P. lunariifolia, P. bourgaei, P. longifolia var. longifolia, P. grandiflora var. grandiflora and P. grandiflora var. fimbrilligera. The second group comprised 2 species, P. leucophracta and P. chimerae. Species-specific bands were observed for P. lycia, P. leucophracta, P. lunariifolia, P. bourgaei, P. chimerae and P. longifolia var longifolia.

Evaluation of antibacterial activities of some medicinal plants from north-West Iran

Aim of the present study was to screen the antibacterial activities of some medicinal plants extracts traditionally used in Azarbaijan area [Iran]. Thirty-six extracts obtained from different parts of ten plants including Tanacetum balsam ita L. [Copmositae], Muscari caucasicum Baker [Hyacinthaceae], Equisetum arvense L. [Equisetaceae], Achillea millefollum L. [Copmositae], Stachys fruticulosa M. Bieb. [Labiatae], Stachys schtschegleevii Sons. ex, Grossh. [Labiatae], Salvia sahendica Boiss and Buhse [Labiatae], Phlomis caucasica Rech. f. [Labiatae], Etchium italicum L. [Boraginaceae] and Thalictrum minus L. [Ranunculaceae] from north-west Iran with traditional medicinal use were examined for their antibacterial activities against some Gram-negative strains such as Escherichia coli, Pseudomonas aeruginosa, Salmonella paratyphi and Serratia marcescens, also, Gram-positive strains of Staphylococcus aureus, Micrococcus luteus, Staph. epidermidis, Streptococcus pneumoniae and Bacillus cereus. The filter paper disc diffusion method as well as broth serial dilution technique were applied to screen the antibacterial efficacy of the extracts and determination of minimum inhibitory values. Results indicated that the majority of tested plant extracts had antibacterial activity at least against one of the selected bacteria, with the exception of Muscari caucasicum. Methanol extract of the aerial part of Thalictrum minus L. [Ranunculaceae] showed the most potent antibacterial activity against Staph. Aureus with MIC value of 0.3125 mg/ml. The results of this study show that most of the studied plants are potentially a good source of antimicrobial agents and support the traditional applications of some of the tested plants

Chemical constituents from rhizome of Phlomis umbrosa / 药学学报

Phlomis umbrosa is a traditional medicinal plant, distributed in the north of China. In the west of Hubei province, its roots were used in the treatment of the rheumatic diseases in Tujia nationality. To study the chemical constituents from the rhizome of Phlomis umbrosa chemical constituents were isolated from the plant by using repeated silica gel, toyopearl HW-40 and preparative HPLC chromatography. The structures of the compounds were elucidated on the basis of one and two dimensional NMR spectroscopic techniques and HRESI-MS. Ten compounds, 6"-syringyl-sesamoside (1), decaffeoylverbascoside (2), calcelarioside B (3), verbascoside (4), isoverbascoside (5), alyssonoside (6), sesamoside (7), shanzhiside methyl ester (8), 8-acetyl-shanzhiside methyl ester (9), 7-epiphlomiol (10) were isolated from P. umbrosa. Compound 1 is a new compound. Compounds 2-6 are isolated from this plant for the first time.

Studies on chemical constituents in root of Phlomis medicinalis I / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in root of Phlomis medicinalis.</p><p><b>METHOD</b>The compounds were isolated and repeatedly purified on macroporous resin, silica gel column chromatography, TLC and Prep-HPLC and the structures were elucidated by physico-chemical properties and NMR spectra.</p><p><b>RESULT</b>Eight compounds were obtained and elucidated as 5-Hydroxy-7-methoxy-4, 6-dimethylphthalide (1), 4-hydroxymethyl-2-furaldehyde (2) and six iridoid glucosides: 6-O-acetyl-shanzhiside methyl ester (3), 8-O-acetyl-shanzhiside methyl ester (4), shanzhiside methyl ester (5), sesamoside (6), phloyoside II (7) and dehydropentstemoside (8).</p><p><b>CONCLUSION</b>All the compounds were isolated from the plant for the first time and 1 and 3 were obtained from the plants of Phlomis for the first time.</p>