Phytochemical analysis and antibacterial/antifungal activity of the essential oil of Phlomis olivieri Benth in Iran.

Phlomis olivieri Benth. is an aromatic plant endemic to Iran belonging to the Lamiaceae family. It is used to treat pain, stomach ache and common cold in Iranian traditional medicine. P. olivieri also has valuable biological properties including antioxidant, antimicrobial and analgesic ones. This was the first study designed to assess the quality, quantity and antimicrobial activity of Phlomis olivieri Benth. essential oil (POEO). Samples were randomly collected from flowering twigs of this species in three locations between Azeran and Kamoo in Kashan, Iran at peak flowering in June 2019. Water distillation extraction was used to obtain the POEO the quantity of which was calculated by weight. Gas chromatography coupled to mass spectrometry (GC/MS) was also used for POEO qualitative analysis, which revealed its chemical compounds and their percentages. Antimicrobial activity of POEO was also determined using the agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) were also measured using the broth microdilution method. The results of the quantitative and qualitative analysis showed that the POEO yield was ~ 0.2292% and its main chemical compounds included the sesquiterpenes germacrene D (26.43%), ß-caryophyllene (20.72%), elixene (6.58%), ß-trans-farnesene (6.17%), ß-Cyclogermacrane (5.04%), germacrene B (4.73%), α-humulene (4.22%), and monoterpene α-pinene (3.22%). The agar diffusion method demonstrated the highest antimicrobial activity of POEO (MIC ~ 14.50 mm) was against the Gram-positive species Streptococcus pyogenes. The POEO also showed the strongest inhibitory and lethal activity against the gram-negative bacterial species Pseudomonas aeruginosa (MIC < 62.50 µg/mL) and S. paratyphi-A (MIC < 62.50 µg/mL and MBC = 125 µg/mL), and fungal species Candida albicans (MIC and MBC = 250 µg/mL) as compared to control-positive antibiotics. Therefore, POEO is a valuable natural alternative rich in sesquiterpenes with strong antimicrobial and antifungal activities against some fungal and bacterial strains. It can also be used in the pharmaceutical, food and cosmetic industries.

Iridoid Glycosides from Phlomis Medicinalis Diels: Optimized Extraction and Hemostasis Evaluation.

Phlomis medicinalis Diels, an important perennial herbal plant unique to the Qinghai-Tibet Plateau, is often used as Tibetan Materia Medicine Radix Phlomii for the treatment of cold, cough, and convergence trauma. In order to efficiently extract the iridoid glycosides from P. medicinalis, an ultrasound-assisted deep eutectic solvent extraction technique was employed. The main parameters influencing the extraction process were studied through single-factor tests and the extraction was optimized by using response surface methodology. The hemostasis activity of total iridoid glycosides (TIG) from P. medicinalis was evaluated in vitro and in mice. The optimization results revealed that the optimal process parameters were liquid-solid ratio 20 : 1, choline chloride-lactic acid concentration 79 %, and sonication time 34 min, under which a TIG extraction yield of 20.73 % was obtained. Meanwhile, high-performance liquid chromatography-photodiode array/mass spectrometry (HPLC-PDA/MS) was employed to characterize the optimized extract and indicated that TIG from P. medicinalis mainly consisted of sixteen reported iridoid glycosides with a total content of 91.22 %. The experimental results in vivo and in vitro indicated that TIG from P. medicinalis had strong hemostasis activities, which may be achieved by increasing the fibrinogen levels. Therefore, the ultrasound-assisted deep eutectic solvent extraction is an effective method to extract iridoid glycosides from P. medicinalis and they will be promising candidates to be developed for medical hemostasis agents.

Anti-asthmatic effects of Phlomis umbrosa Turczaninow using ovalbumin induced asthma murine model and network pharmacology analysis.

BACKGROUND: Phlomis umbrosa Turczaninow has been used as a tradition herbal medicine for treating various inflammatory diseases. PURPOSE: In present study, we explored the effects of P. umbrosa on asthma induced by ovalbumin (OVA) and elucidated the mechanism via in vivo verification and network pharmacology prediction. METHODS: The animals were intraperitoneally injected OVA on day 1 and 14, followed by OVA inhalation on days 21, 22, and 23. The animals were daily treated P. umbrosa extract (PUE, 20 and 40 mg/kg) by oral gavage from day 18 to day 23. RESULTS: PUE significantly decreased airway hyperresponsiveness, eosinophilia, and the production of inflammatory cytokines and OVA specific immunoglobulin E in animals with asthma, along with a reduction in airway inflammation and mucus secretion in lung tissue. In network analysis, antiasthmatic effects of PUE were closely related with suppression of mitogen-activated protein kinases and matrix metalloproteinases (MMPs). Consistent with the results from network analysis, PUE suppressed the phosphorylation of ERK and p65, which was accompanied by a decline in MMP-9 expression. CONCLUSION: Administration of PUE effectively reduced allergic responses in asthmatic mice, which was associated with the suppressed phosphorylation of ERK and p65, and expression of MMP-9. These results indicate that PUE has therapeutic potential to treat allergic asthma.

Chemical profiling, antiviral and antiproliferative activities of the essential oil of Phlomis aurea Decne grown in Egypt.

Here, we investigated the chemical composition of the edible Phlomis aurea oil and its anticancer potential on three human cancer cell lines, as well as its antiviral activity against Herpes simplex-1 (HSV-1). Exploring Phlomis aurea Decne essential oil by gas chromatography coupled with mass spectrometry (GC/MS) revealed the presence of four major components: germacrene D (51.56%), trans-ß-farnesene (11.36%), α-pinene (22.96%) & limonene (6.26%). An antiproliferative effect, as determined by the MTT assay, against human hepatic, breast and colon cancer cell lines, manifested IC50 values of 10.14, 328.02, & 628.43 µg mL-1, respectively. Cytotoxicity assay of the Phlomis oil against Vero cell lines revealed a safe profile within the range of 50 µg ml-1. Phlomis essential oil induced the apoptosis of HepG2 cells through increasing cell accumulation in sub G1 & G2/M phases, decreasing both S & G0/G1 phases of the cell cycle, triggering both caspases-3 &-9, and inhibiting cyclin dependent kinase-2 (CDK2). The antiviral activity of the oil against HSV-1 was investigated using the plaque reduction assay, which showed 80% of virus inhibition. Moreover, the molecular docking in silico study of the four major chemical constituents of the oil at the CDK2 binding site demonstrated marked interactions with the ATP-binding site residues through alkyl & Pi-alkyl interactions. Cell cycle distribution of HepG2 cells was studied using flow cytometry to highlight the apoptotic mechanistic approaches by measuring caspases-3 &-9 and CDK2 activities. Thus, the edible Phlomis oil can be regarded as a candidate for in vivo studies to prove that it is a promising natural antiviral/anticancer agent.

Anti-proliferative effects of Ziziphus spina-christi and Phlomis russeliana leaf extracts on HEK293 and MCF-7 Cell Lines and Evaluation of Bax and Bcl-2 Genes Expression Level in MCF-7 Cells.

To investigate the effects of Phlomis russeliana and Ziziphus spina-christi leaf extracts on apoptosis in breast cancer MCF-7 cells. Cell lines were divided into a control group and the groups exposed to 0.001, 0.01, 0.1, 1, and 10 mg/ml of Ziziphus spina-christi and Phlomis russeliana leaf extracts. Cell viability was quantified by the MTT assay. The expression of Bax and Bcl-2 genes was evaluated by Real-time PCR analysis. Statistical analysis was performed using ANOVA. HEK293 cell viability significantly increased in the groups exposed to 0.001, 0.01, and 0.1 mg/ml of Z.christi leaf extract and decreased in the group exposed to 10 mg/ml of P.russeliana leaf extract. MCF-7 cells viability significantly decreased in the groups exposed to 0.001, 0.01, 0.1, 1 and 10 mg/ml of Z.christi leaf extract and increased in the groups exposed to 0.001 and 0.01 mg/ml of P.russeliana leaf extract. The exposure of MCF-7 cells to 1 and 10 mg/ml of P.russeliana leaf extract also led to a significant decrease in cell viability. The cytotoxic effect of Z.christi was higher than P.russeliana leaf extracts on MCF7 cells.  1 mg/ml of Z.christi leaf extracts also significantly increased the expression level of Bax and Bcl-2 genes in MCF7 cells. Bcl-2 gene expression significantly increased in the group exposed to 10 mg/ml of P.ruseliana leaf extract.Despite P.russeliana leaf extract, lower Z.christi leaf extract concentrations inhibited MCF-7 cells proliferation. Ziziphus spina-christi and phlomis russeliana leaf extracts mechanism of action has occurred through the Bax-independent apoptotic pathway on MCF-7 cells.

A comparative study on chemical composition and antimicrobial activity of essential oils from three Phlomis species from Uzbekistan.

Essential oils obtained from the aerial parts of Phlomis bucharica, P. salicifolia and P. sewerzowii were determined using GC-FID and GC-MS methods. A total of 76 components were identified in the three species representing 97.12, 88.34, and 96.41% of the whole oil, respectively. High percentages of thymol (20.41%) and camphor (14.46%) exist in P. bucharica oil. Methyl palmitate predominates in P. salicifolia oil representing 51.15% whereas thymol (35.76%) is the major constituent in P. sewerzowii essential oil. GC-MS analyses showed that P. bucharica and P. sewerzowii are more closely related comparable to P. salicifolia. The antimicrobial activity of the essential oils was assessed against different microorganisms using agar-disc diffusion and broth microdilution assay. Among the three tested species, the essential oil of P. salicifolia showed the highest antibacterial activity.

Anti-inflammatory, analgesic and in vivo-in vitro wound healing potential of the Phlomis rigida Labill. extract.

ETHNOPHARMACOLOGICAL RELEVANCE: The preparations of Phlomis aerial parts are used traditionally in Anatolia for wound healing and in inflammatory disorders. METHODS: For the identification of the active fraction, the air dried aerial parts of Phlomis rigida Labill. were extracted by methanol and fractionated successively by n-hexane, dichloromethane and ethyl acetate, respectively. The phenolic constituents were characterized by the Folin-Ciocaltheu method; the antioxidant activity was performed by ABTS and DPPH radical scavenging assays. In vitro anti-inflammatory activity was evaluated by LOX enzyme inhibition, spectrophotometrically as well as cell cultures. The wound healing properties of P. rigida extract gels were studied via in vitro cell culture methods and in vivo by excisional wound model using Balb-c mice. The P. rigida extract was analyzed and characterized by GC-FID, GC-MS, and LC-MS. RESULTS: The P. rigida methanol extract showed moderate LOX inhibitory at IC50 = 19.5 ± 2.8 µg/mL whereas the antioxidant activity was by DPPH• IC50 = 0.89 mg/mL, and by ABTS• IC50 = 0.99 mg/mL, respectively. In addition, a remarkable P. rigida extracts anti-inflammatory activity was observed in the cell culture assay, which was then confirmed by the in vitro wound healing activity applied at 0.125-0.5 mg/mL concentrations, resulting in a dose-dependent increase in wound closure at the final stage. The P. rigida gel formulation was prepared to evaluate the extract in vivo, whereas the experimental results of the new gel formulation supported the findings of the in vitro wound healing activity. CONCLUSION: The findings of this in vitro and in vivo study suggest that the wound healing and anti-inflammatory properties provide a scientific evidence of the ethnopharmacological application of Phlomis species.

Iridoid glucosides and triterpene acids from Phlomis linearifolia, growing in Uzbekistan and its hepatoprotective activity.

A number of iridoids and triterpene acids, such as pulchelloside, sesamoside, shanshiside methyl ester, barlerin, gypsogenin acid and acetate of gypsogenic acid were isolated from the aerial part of Phlomis linearifolia and their structures were confirmed by NMR, mass and IR spectroscopy. In addition, the hepatoprotective potential of iridoid fraction from P. linearifolia aerial parts was tested against CCl4 induced fibrosis in rats. The iridoid fraction not only prevented the manifestation of the hepatotoxic effect of CCl4, but rather quickly eliminated the effects of developing intoxication. The hepatoprotective activity of the SI was confirmed to be effective and exceeds knows drug carsil.Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2019.1677650.

Protective effects of Phlomis umbrosa extract on a monosodium iodoacetate-induced osteoarthritis model and prediction of molecular mechanisms using transcriptomics.

BACKGROUND: Phlomis umbrosa Turczaninow root has been traditionally used to treat fractures, rheumatoid arthritis, and arthralgia. However, the effects and mechanisms of P. umbrosa on osteoarthritis (OA) remain poorly understood and a functional genomic approach has not been investigated. AIM: The purpose of this study was to investigate the effects and mechanisms of P. umbrosa extract (PUE) on OA using transcriptomic analysis. METHODS: We performed joint diameter measurements, micro computed tomography, and histopathological analysis of monosodium iodoacetate (MIA)-induced OA rats treated with PUE (200 mg/kg) for 3 weeks. Gene expression profiling in articular cartilage tissue was then performed using RNA sequencing (RNA-seq) followed by signaling pathway analysis of regulatory genes. RESULTS: PUE treatment improved OA based on decreased joint diameter, increased joint morphological parameters, and histopathological features. Many genes involved in multiple signal transduction pathway and collagen activation in OA were differentially regulated by PUE. These included genes related to Wnt/ß-catenin, OA pathway, and sonic hedgehog signaling activity. Furthermore, PUE treatment downregulated cartilage damage factors (MMP-9, MMP-13, ADAMTs4, and ADMATs5) and upregulated chondrogenesis (COL2A1 and SOX-9) by regulating the transcription factors SOX-9, Ctnnb1, and Epas1. CONCLUSION: Based on the results of gene expression profiling, this study highlighted the molecular mechanisms underlying the effects of PUE in MIA-induced OA rats. The findings provide novel insight into the mechanisms by which PUE treatment-induced gene expression changes may influence OA disease progression. Taken together, the results suggest that PUE may be used as a source of therapeutic agents for OA.

In Vivo Wound Healing and In Vitro Anti-Inflammatory Activity Evaluation of Phlomis russeliana Extract Gel Formulations.

The air-dried aerial parts of Phlomis russeliana (Sims) Lag. Ex Benth. was extracted by methanol and fractionated by n-hexane, dichloromethane, and ethyl acetate, respectively. The wound healing properties of P. russeliana extract gel was evaluated using the in vivo excisional wound model using Balb-c mice. Initially, the P. russeliana methanol extract showed LOX inhibitory activity at IC50 = 23.2 µg/mL, whereas the DPPH• assay showed IC50 = 0.89 mg/mL, and the ABTS• assay showed IC50 = 0.99 mg/mL, respectively. In addition, a remarkable anti-inflammatory activity was observed in the cell culture assay. Thereafter, activity-guided fractionation was performed by LOX enzyme inhibition assays, and the structures of the two most active fractions were revealed by both GC-FID and GC/MS analyses, simultaneously. Phytol and 1-heptadecanoic acid were characterized as the active constituents. Moreover, the P. russeliana extract gel formulation was applied for in vivo tests, where the new gel formulation supported the in vitro anti-inflammatory activity findings. As a conclusion, this experimental results support the wound healing evidence based on the ethnobotanical application of Phlomis species with further potential.

Chemical composition and antioxidant activity of Phlomis leucophracta, an endemic species from Turkey.

The chemical composition, and antioxidant activity of the essential oils of the endemic Phlomis leucophracta P. H. Davis et Hub.-Mor. was investigated. The major compounds of the essential oil were linalool (36.4%), spathulenol (8.4%) and caryophyllene oxide (8.4%). The composition of the oil differs with published data, suggesting other chemotype. Moreover, the oil of this species was analysed for its antioxidant activity for the first time and results indicate it possess strong antioxidant activity comparable with already known antioxidants, such as ascorbic acid, BHT, or Trolox. Presented results suggest that this endemic species has strong potential to be used in food and pharmacological industries, and therefore they need to be further investigated.

Vasodilatory effect and structural-activity relationship of a group of iridoid glucosides from Phlomis likiangensis.

As a folk medicine, Phlomis likiangensis is traditionally used in China to activate collaterals and protect cardiovascular system. We hypothesized that the beneficial effects of Phlomis likiangensis may be related to vasodilatation. In the present study, twelve known iridoid glucosides (1-12) were isolated from Phlomis likiangensis. The vasodilatory effects and the underlying mechanisms of the main components (iridoid glucosides) of Phlomis likiangensis on rat aortic rings were investigated. The result showed that iridoid glucosides significantly increased the vasodilatation in rat aortic rings, which was abolished by removing the endothelium of the vessels or by eliminating the generation of nitric oxide. Finally, the structure-activity relationship of compounds 1-12 was also speculated. Our findings provide the first evidence that the iridoid glucosides of Phlomis likiangensis may be the pharmacodynamic basis for its traditional efficacy.

Evaluating antimicrobial and antioxidant capacity of endemic Phlomis russeliana from Turkey and its antiproliferative effect on Human Caco-2 Cell Lines.

In this study, the antimicrobial, antioxidant and antitumor activity of ethanol extracts obtained from Phlomis russeliana (Sims.) Lag. ex Benth. (Lamiaceae) were evaluated. Disc diffusion and microdilution methods were used to test the extracts for antimicrobial activity against seven bacteria strains (Bacillus cereus ATCC 7064, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 6538P, Escherichia coli ATCC 10538, Proteus vulgaris ATCC 6899, Salmonella typhimurium CCM 5445 and Pseudomonas aeruginosa ATCC 27853) and four yeast strains (Kluyveromyces fragilis ATCC 8608, Rhodotorula rubra ATCC 70403, Debaryomyces hansenii DSM 70238 and Candida albicans ATCC 10239). Notably, they were more effective against the yeast strains than the bacterial strains. Of the yeast cultures, D. hanseii was among the most susceptible, having an inhibition zone of 16.2 mm with minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) of 64(128)µg/ml, respectively. For cytotoxic determination, Caco-2 cells were cultured as per ATCC protocol, and were treated with log concentrations (5-80 mg/ml) of P. russeliana. The potency of cell growth inhibition for each extract was expressed as an IC50 value. Moreover, oxidant capacity was evaluated via TOC assay. This product induced antiproliferative activity of 31.33% at 40 mg/ml and 20.96% at 80 mg/ml, without toxic effects on cells, although the oxidant capacity was decreased to 27.06 ± 0.7 nm in the 80 mg/ml-applied group compared to 47.9 ± 1.8 nm in the untreated one. Advanced pharmacological studies are needed to further evaluate P. russeliana for distinctive features.

Protective effects induced by alcoholic Phlomis fruticosa and Phlomis herba-venti extracts in isolated rat colon: Focus on antioxidant, anti-inflammatory, and antimicrobial activities in vitro.

Phlomis fruticosa L. and P. herba-venti are species belonging to the Lamiaceae family, which have been traditionally used to prepare tonic and digestive drinks. Multiple studies also demonstrated the inhibitory effects of P. fruticosa extracts and essential oil against oxidative/proinflammatory pathways and bacterial strains deeply involved in ulcerative colitis. Considering these findings, the present study evaluated the effects of alcoholic P. fruticosa and P. herba-venti leaf extracts in isolated rat colon challenged with Escherichia coli lipopolysaccharide (LPS), an ex vivo experimental paradigm of ulcerative colitis. In this context, we assayed colon levels of pro-oxidant and proinflammatory biomarkers, including nitrites, malondialdehyde (MDA), lactate dehydrogenase (LDH), and serotonin (5-HT). Additionally, the extracts have been tested in order to evaluate possible inhibitory effects on specific bacterial and fungal strains involved in ulcerative colitis. Alcoholic P. fruticosa and P. herba-venti extracts were able to blunt LPS-induced nitrite, MDA, 5-HT, and LDH levels in isolated rat colon. The same extracts also inhibited the growth of Pseudomonas aeruginosa, E. coli, Staphylococcus aureus, Candida albicans and Candida tropicalis. In conclusion, our findings show a potential role exerted by alcoholic P. fruticosa and P. herba-venti in managing the clinical symptoms related to ulcerative colitis.

Chromatographic analysis, antimicrobial and insecticidal activities of the essential oil of Phlomis floccosa D. Don.

The present study was conducted to investigate for the first time the chemical composition and insecticidal and antimicrobial properties of Phlomis floccosa D. Don. essential oil isolated by hydro-distillation of its aerial parts. Qualitative and quantitative analyses of the essential oil on the basis of gas chromatography and mass spectrometry (GC/FID and GC/MS) revealed the presence of 59 organic volatiles representing 86.9% of the total constituents. The major compounds were germacrene-D (19.7%) followed by ß-caryophyllene (15.5%), caryophyllene oxide (8.3%), hexadecenoic acid (7.9%) and carvacrol (6.1%). The broth microdilution method was conducted to test the antimicrobial activity of the essential oil against five pathogens: Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, Bacillus subtilis and the yeast Candida albicans. The oil of P. floccosa D. Don. shows promising activity against the Gram-positive bacteria S. aureus and B. subtilis and the yeast C. albicans (minimum inhibitory concentration = 625 µg.mL-1 ). The effectiveness of the essential oil was assessed on both Tribolium confusum and Sitophilus zeamais pest insects and it was more pronounced against T. confusum, showing a moderate mortality of 47.5%.

Phenylethanoid glycosides of Phlomis younghusbandii Mukerjee ameliorate acute hypobaric hypoxia-induced brain impairment in rats.

High altitude cerebral edema (HACE), whose development process is associated with oxidative stress and inflammatory response, is a life-threatening condition caused by rapid ascent speed to high altitudes. Phenylethanoid glycosides (PhGCs) are primary active constituents isolated from Phlomis younghusbandii Mukerjee that reportedly exhibit potent anti-oxidant and anti-inflammatory activities. The present study aims to investigate the protective effect of phenylethanoid glycosides (PhGCs) from P. younghusbandii in acute hypobaric hypoxia (AHH) - stimulated HACE rats and its underlying mechanisms. The expression of pro-inflammatory cytokine levels (IL-1ß, TNF-α, and IL-6) was detected by RT-PCR and ELISA at mRNA and protein levels in brain tissues. Western blotting was carried out to measure the major protein levels (IL-1ß, TNF-α, and NF-κB) in brain tissues. The oxidative stress biomarkers (MDA, SOD, and GSH) were evaluated using kits. Results demonstrate that PhGCs significantly improved pathological changes in brain tissues, reduced the brain's water content, and attenuated the production and mRNA expression of pro-inflammatory cytokines. Furthermore, the increased oxidative stress and the decrease in anti-oxidant stress system under the AHH condition were also abrogated reversely through PhGCs treatment by elevating the levels of SOD and GSH and suppressing the accumulation of MDA. Simultaneously, there was also a significant reduction in NF-κB, IL-1ß, and TNF-α protein expression levels in brain tissues, suggesting that blocking the NF-κB signaling pathway activation prevented the production of pro-inflammatory cytokines. Taken together, these findings indicate that PhGCs may afford a protectively intervene in HACE through the suppression of oxidative stress and inflammatory response via the inhibition of the NF-κB signaling pathway, indicating that PhGCs are promising agents for the treatment of acute HACE.

Investigation of the synergistic effects of paclitaxel and herbal substances and endemic plant extracts on cell cycle and apoptosis signal pathways in prostate cancer cell lines.

Paclitaxel, which isolated from Taxus brevifolia, is recently started to be used against prostate cancer treatment and it is a very effective compound against cancer. In this study, we aimed to test the synergistic effect of two plant active compounds (sulphoraphane (SFN) and silymarin (SILY)) and several endemic plant species from Turkey (such as Phlomis leucophracta, Rubia davisiana, Alkanna tinctoria), which are known to have anticarcinogenic effect on androgen-independent PC3 and DU145, and androgen-dependent VCaP prostate cancer cell lines, with paclitaxel on the expression of cell cycle signaling and apoptosis regulator genes. Herbal substances and endemic herbal extracts were combined with Paclitaxel drug. IC50 doses were identified as real-time online. The most effective synergistic doses were determined according to isobologram analysis. The apoptotic effects of effective combined doses were evaluated by TUNEL, Annexin V, and JC-1 methods. Apoptotic and/or cell cycle arrest effects of confirmed combined doses on the expression of genes in these pathways were assessed by real-time online. Endemic plant extracts (Alkanna tinctoria, Phlomis leucophracta and Rubia davisiana, IC50 < 220 µg/ml) and herbal substances (SILY, and SFN IC50 < 130 µM) indicated antiproliferative and apoptotic effects in prostate cancer cell lines. They testified to the synergistic effect of paclitaxel with endemic plant extracts (Combination Index CI, ED50 < 0.41). The combinations, which indicate the synergistic effect was increased to the Bax/Bcl­2 ratio by suppressing Bcl­2 gene expression into the prostate cancer cell lines. Besides, they increased the expression of TNFRSF10A, TNFRSF1A, CHEK1, CDKN1A, CDKN2B, CDK8, CDKN3 and CASP14 and decreased BAD, CDK5RAP1, CDC20, cyclin H, CDK5RAP1, CDC20. The effective doses of paclitaxel were reduced and G2/M arrest was induced by the endemic plant extracts and herbal substances that indicate a synergistic effect with paclitaxel. By using different combination of herbal extracts or active substances with paclitaxel, more economical and efficient treatment strategies can be developed.

Recent Updates on the Phytochemistry and Pharmacological Properties of Phlomis viscosa Poiret.

Crude ethanolic extracts from Phlomis viscosa Poiret leaves from the Judea region (Israel) are renowned for their remarkable geroprotective properties: anti-inflammatory, anti-diabetic, and anti-cancer. A phytochemical investigation carried out in this study revealed that the tested plant might belong to a particular distinct chemotype because its phytochemicals are different from compounds that were mentioned in the literature. Among the compounds identified by us was diosmin, the synthetic derivatives of which were further obtained and investigated. In particular, activities of the isolated compounds and synthesized diosmin derivatives were assessed. Our results revealed that the following compounds significantly lessened secretion of some pro-inflammatory cytokines: diosmin, himachala-2-diene, and 5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl) chromen-4-one. In addition, diosmin, synthesized diosmin derivatives, and some identified terpenes were found to have anti-diabetic activities. A significant anti-cancer effect of the whole extract on U-87 (human glioblastoma carcinoma cells line) and MCF7 (human breast carcinoma cell line) was also demonstrated, and it was better than that of DOX (doxorubicin). Collectively, the results obtained in the in vitro models suggest a wide spectrum of beneficial bioactivities of the extract and its active compounds.

[A new isobenzofuranone derivative from Phlomis betonicoides and its antibacterial activity].

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-2H-furo[3,4-h]chromen-7(9H)-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].

Flavonoid Constituents of Phlomis (Lamiaceae) Species Using Liquid Chromatography Mass Spectrometry.

INTRODUCTION: Phlomis is one of the medicinal genera of Lamiaceae. This genus has unique medicinal properties. Consequently, appropriate methods need to be described for the identification of the chemical compounds. OBJECTIVE: A liquid chromatography tandem mass spectrometry (LC-MS/MS) technique on a triple quadrupole mass spectrometer (TQMS) was used for separation and identification of leaf flavonoid compounds for seven Phlomis species including Phlomis kurdia, Ph. aucheri, Ph. olivieri, Ph. bruguieri, Ph. persica, Ph. anisodonta and Ph. elliptica. METHODOLOGY: The flavonoid solution of air-dried leaves (10.5 g) was extracted using 85% methanol. The chromatogram was treated with three systems: methanol-water, chloroform-methanol and acetic acid. The extracts were analysed using LC-MS/MS. The MS2 detection was performed under negative mode electrospray ionisation (ESI). The identification of constituents was based on authentic references used in the identification process. RESULTS: A total of 35 chemical compounds were detected from which 32 were identified as flavonoids through comparison with published literature and reference standards. These compounds were distributed in four flavonoid classes. Flavones (12), flavonols (11), flavanones (8) and flavane (1) were the main groups appearing in almost all of the studied samples. The flavonoids such as naringenin, chrysoeriol, eriodictyol, dimethoxyflavanone, apigenin, luteolin, kaempferol and rhamnetin were in high proportions. Moreover, 22 flavonoid compounds were first reported in this study for Phlomis species. CONCLUSION: The fragmentation patterns of the compounds during collision induced dissociation (CID) clarified information of the compounds analysed. The detailed flavonoid compositions of Phlomis species provide the appropriate context from phytochemical and phytotherapeutics points of view. Copyright © 2017 John Wiley & Sons, Ltd.