Effect of ultrasound-assisted cold plasma pretreatment on cell wall polysaccharides distribution and extraction of phenolic compounds from hyssop (Hyssopus officinalis L.).

Hyssopus officinalis L (Hyssop) is a good source of phenolic compounds. However, conventional methods for extraction of these compounds typically take a long time and have relatively low recovery rates. This study focused on cold atmospheric plasma (CAP) pretreatment and investigated its effects on the ultrasound-assisted extraction (UAE) of phenolic compounds from hyssop. Hyssop was treated at dielectric barrier discharge plasma with air and nitrogen gases for 5, 10, and 15 min. Optical emission spectroscopy was used to evaluate present active species in the plasma. The water contact angle changes, cell wall polysaccharides distribution, and structural variations of the treated samples were determined after treatment. Antioxidant activity and total phenolic contents (TPC) of the extracts were also evaluated. The results showed that CAP treatment reduced the contact angle making surface more hydrophilic. Compared with hyssop, overall no significant changes in the basic structure of all treated samples or the formation of new functional groups were recognized. In addition, CAP pretreatment before UAE increased the antioxidant activity of extracts according to the FRAP assay than the un-pretreated sample and conventional solvent extraction method. Also, TPC increased in samples treated with nitrogen plasma.

Monoterpenoid derivatives from Hyssopus cuspidatus Boriss. and their bioactivities.

Six undescribed monoterpenoids, together with twelve known compounds were isolated and identified from Hyssopus cuspidatus Boriss. Their structures were established by spectroscopic analysis, and the absolute configurations were established by ECD calculations and single-crystal X-ray diffraction crystallography. The isolated compounds were tested for their anti-inflammatory, antibacterial and antitumor activities. Most of the compounds showed potent anti-inflammatory activities. Among them, 3ß-hydroxy-7,8-dihydro-ß-ionone (8), oleanolic acid (17) and acetylpleamolic acid (18) showed strong anti-inflammatory activity against IL-6 and TNF-α in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Several compounds showed moderate inhibitory activities against Staphylococcus aureus, Candida albicans, and Escherichia coli. And (4S)-p-menth-l-ene-7,8-diol 8-O-ß-D-glucopyranoside (16) showed antitumor activities against MCF-8 and HT-29 cell lines with IC50 values of 93.39 ± 3.69 and 71.89 ± 2.94 µM, respectively. Oleanolic acid (17) showed moderate antitumor activity against HT-29 cell lines with an IC50 value of 52.62 ± 1.63 µM. In this study, the discovery of anti-inflammatory, antibacterial and antitumor components from H. cuspidatus could benefit further development and utilization of this plant.

The extract from Hyssopus cuspidatus Boriss. Prevents bronchial airway remodeling by inhibiting mouse bronchial wall thickening and hASMC proliferation and migration.

ETHNOPHARMACOLOGICAL RELEVANCE: Bronchial asthma, a non-communicable chronic respiratory disease, affects people of all ages. An important pathological feature of bronchial asthma is airway remodeling. Hyssopus cuspidatus Boriss. has been used to treat bronchial asthma for over 100 years in Uygur medicine. The ethanol extract of Hyssopus cuspidatus Boriss.(JAX2) can improve airway inflammation in asthma. However, the anti-asthmatic airway-remodeling effect of JAX2 is unclear. AIM OF THE STUDY: The current study investigated the anti-airway remodeling effect of JAX2 and elucidated its mechanism of action. MATERIALS AND METHODS: The present study established an ovalbumin-induced mouse model of asthma and platelet-derived growth factor-BB-induced human airway smooth muscle cells (hASMCs) proliferation model, with dexamethasone (DEX) and feining tablets (FNP) designated as positive control drugs. Pathological changes in lung tissues were observed using hematoxylin and eosin staining. Interleukin (IL)-5, IL-10, IL-13, and IL-33 levels in the bronchoalveolar lavage fluid (BALF) and serum of mice were determined using enzyme-linked immunosorbent assay (ELISA). Changes in the expression and distribution of TGF-ß1, p-ERK1/2, Smad2/3, and p-Smad3 in lung tissues were determined using immunohistochemistry. Western blotting (WB) was used to determine the protein levels of p-ERK1/2 in lung tissues and cells. MTS assay was used to determine the effects of JAX2 on cell proliferation. IL-5, IL-10, IL-13, MMP-2, and MMP-9 levels in the cell supernatant were determined using ELISA. HASMCs migration was observed using the scratch and transwell methods. The effect of JAX2 on the hASMCs cycle was determined using flow cytometry. RESULTS: JAX2 significantly improved the pathological status of lung tissues in asthmatic mice. It could also significantly reduce IL-5, IL-13, and IL-33 levels in the BALF and serum of asthmatic mice in a dose-dependent manner and significantly increase IL-10 levels. TGF-ß1, p-ERK1/2, Smad2/3, and p-Smad3 expression in lung tissues were decreased in a dose-dependent manner. The protein level of p-ERK1/2 in lung tissues was also reduced. JAX2 could significantly inhibit the proliferation and migration of PDGF-BB-induced hASMCs. IL-5, IL-13, MMP-9, and MMP-2 levels decreased significantly, and IL-10 levels increased significantly in a dose-dependent manner in the cell supernatant. JAX2 could block hASMCs in the G0/G1 phase, thereby inhibiting cell proliferation. p-ERK1/2 protein levels were found to decrease in a dose-dependent manner. CONCLUSIONS: JAX2 significantly inhibits airway remodeling in asthma. Its mechanism of action may be inhibiting the proliferation and migration of hASMCs, releasing inflammatory factors and metalloproteinases, activating the ERK1/2 signal pathway, and promoting the secretion of anti-inflammatory factors.

A new macrocyclic spermidine alkaloid from the aerial part of Hyssopus cuspidatus Boriss.

Hyssopus cuspidatus Boriss. grows in Xinjiang, China. A new macrocyclic spermidine alkaloid, namely hyssopusizine (1), along with sixteen known compounds were isolated and identified from the aerial parts of H. cuspidatus. Their structures were elucidated on the basis of spectroscopic data and comparison with the literature. Among them, fifteen compounds were isolated from H. cuspidatus for the first time. The absolute configuration of compound 1 was established by comparing the calculated and experimental ECD spectroscopic data. All isolated compounds were tested for their antioxidant and antimicrobial activities. Among them, compound 10 exhibited significant effects on ABTS free-radical scavenging activity with an IC50 value of 15.6 µM. Compounds 5-7 exhibited potent antioxidant activities against ABTS and DPPH. Most compounds exhibited moderate antimicrobial activities. Hyssopusizine (1) is the first macrocyclic spermidine alkaloid discovered from the Hyssopus genus.

In vitro, in vivo and in silico evaluation of the anti-inflammatory potential of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal properties of hyssop have been used in traditional medicine since ancient times, inter alia, in diseases/conditions with an inherent inflammatory process. AIM OF THE STUDY: Accordingly, the aim of this study was to investigate the anti-inflammatory properties of hyssop herb preparations (essential oil and methanol extracts) in vivo, in vitro and in silico. MATERIALS AND METHODS: For in vitro testing of essential oils and extracts of hyssop herb, the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzyme assays were used. In vivo anti-inflammatory potential of the extracts (at doses of 50, 100 and 200 mg/kg) was assessed using the carrageenan-induced rat paw edema test. Molecular docking and dynamics were used for in silico testing of the inhibitory activity of chlorogenic (CA) and rosmarinic (RA) acids, as the dominant compounds in the tested methanol extracts against COX-1 and COX-2 enzymes. RESULTS: Significant inhibitory activity was shown in the COX-2 test regarding extracts (essential oils did not exhibit any significant activity). Namely, all analyzed extracts, at a concentration of 20 µg/mL, showed a percentage of inhibition of COX-2 enzyme (54.04-63.04%), which did not indicate a statistically significant difference from the positive control of celecoxib (61.60%) at a concentration of 8.8 µM. In vivo testing showed that all methanol extracts of hyssop herb, at the highest test dose of 200 mg/kg in the third and fourth hours, after carrageenan administration, exhibited a statistically significant (p < 0.05) inhibitory effect on the increase in rat paw edema in relation to control. This activity is comparable or higher in relation to the reference substance, indomethacin, at a concentration of 8 mg/kg. The preliminary in silico results suggest that investigated compounds (RA and CA) showed better inhibitory activity against COX-1 and COX-2 than standard non-steroidal anti-inflammatory drug (NSAID), ibuprofen, as evident from the free binding energy (ΔGbind in kJ mol-1). The binding energies of the docked compounds to COX-1 and -2 were found to be in the range between -47.4 and -49.2 kJ mol-1. Ibuprofen, as the one NSAID, for the same receptors targets, showed remarkably higher binding energy (ΔGbind = -31.3 kJ mol-1 to COX-1, and ΔGbind = -30.9 kJ mol-1 to COX-2). CONCLUSION: The results obtained not only support the traditional use of hyssop herb in inflammatory conditions in folk medicine, but also open the door to and the need for further in vivo testing of extracts in order to examine the molecular mechanism of anti-inflammatory activity in living systems and possibly develop a new anti-inflammatory drug or supplement.

Hyssopus Essential Oil: An Update of Its Phytochemistry, Biological Activities, and Safety Profile.

The genus Hyssopus is widespread in central Asia, East Mediterranean, and Mongolian areas. It has six main species which are used as herbal remedies, such as Hyssopus officinalis which is used as a condiment and flavoring agent in food industry. The other five species are H. ambiguus, H. cuspidatus, H. latilabiatus, H. macranthus, and H. seravschanicus. Its species are used in the treatment of various ailments such as cold, cough, loss of appetite, fungal infection, and spasmodic condition. Its constituents especially essential oils are popularly used as an additive in beverages, foods, and cosmetics. The volatile constituents are used for aroma in the food industry, cosmetic industry, and household products. The important active constituents in its essential oils are ß-pinene, pinocamphone, isopinocamphone, and other terpenoids. Hyssopus genus is also bundled with other secondary metabolites including flavonoids luteolin, quercetin, apigenin, and their glucosides, as well as phenolic compounds including ferulic, p-hydroxy-benzoic acid, protocatechuic acid, chlorogenic, and caffeic acid. Combinedly, the extracts of Hyssopus are reported to have potential antiviral and antifungal activities proven using in vitro studies, whereas in vivo investigations have reported the crucial role of Hyssopus extracts in plasma membrane relaxation, cytotoxic, and sedative effects. This plant is believed to be relatively safe at levels commonly used in foods; nevertheless, more studies are needed to determine the safety profile.

Non-volatile compounds of Hyssopus cuspidatus Boriss and their antioxidant and antimicrobial activities.

Hyssopus cuspidatus is a famous spice and an aromatic vegetable. Few information could be available concerning its non-volatile chemical composition and bioactivities. Preliminary bioactive evaluations on the crude ethanol extract and its four fractions disclosed that the ethyl acetate fraction (EAF) exhibited antioxidant and antimicrobial bioactivities. LC-MS/MS analysis of EAF helped to identify sixty-four compounds, and phenolic compounds were the dominant components. Systematic separation and purification of EAF led to the isolation of thirty-four compounds. Six compounds were identified to be new and eighteen compounds were discovered from H. cuspidatus for the first time. Rosmarinic acid, methyl rosmarinate, butyl rosmarinate and salvigenin were the major components of EAF and their contents were determined. Most of isolated compounds exhibited significant or moderate antioxidant and antimicrobial activities. This research supported the edible application of H. cuspidatus and disclosed the potency of it as a natural antioxidant and antimicrobial food additive.

Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation.

It seems quite necessary to obtain effective substances from natural products against inflammatory response (IR) as there are presently clinical problems regarding accompanying side effects and lowered quality of life. This work aimed to investigate the abilities of hyssopuside (HY), a novel phenolic glycoside isolated from Hyssopus cuspidatus (H. cuspidatus), against IR in lipopolysaccharide (LPS)-induced RAW 264.7 cells and mouse peritoneal macrophages. The results indicated that HY could reduce nitric oxide (NO) production and inhibit the production and secretion of pro-inflammatory mediators including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in LPS-stimulated macrophages. Moreover, data from the immunofluorescence study showed that HY suppressed nuclear translocation of nuclear factor-kappa B (NF-κB) upon LPS induction. The Western blot results suggested that HY reversed the LPS-induced degradation of IκB (inhibitor of NF-κB), which is normally required for the activation of NF-κB. Meanwhile, the overexpression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) diminished significantly with the presence of HY in response to LPS stimulation. On the other hand, HY had a negligible impact on the activation of mitogen-activated protein kinase (MAPK) pathways. Moreover, an in silico study of HY against four essential proteins/enzymes revealed that COX-2 was the most efficient enzyme for the interaction, and binding of residues Phe179, Asn351, and Ser424 with HY played crucial roles in the observed activity. The structure analysis indicated the typical characterizations with phenylethanoid glycoside contributed to the anti-inflammatory effects of HY. These results indicated that HY manipulated its anti-inflammatory effects mainly through blocking the NF-κB signal transduction pathways. Collectively, we believe that HY could be a potential alternative phenolic agent for alleviating excessive inflammation in many inflammation-associated diseases.

Comparison of antifungal activity of selected essential oils against Fusarium graminearum in vitro.

INTRODUCTION: Fusaria are microscopic filamentous fungi which are spread in soil, in various organic substrates, and include more than 80 phytopathogenic species which are predominantly hosted by cereals, fruits and vegetables. Many of these species, under certain conditions, are capable of synthesizing secondary metabolites, mycotoxins. At present, various substances are used for their elimination and one of the solutions appears to be essential oils. In the presented study, the antifungal activity of essential oils was researched in vitro. MATERIAL AND METHODS: In this study, two standard fungal isolates Fusarium graminearum CCM F-683 and Fusarium graminearum CCM 8244 (Brno, Czech Republic) were used. The antifungal effect of 6 tested essential oils (Syzygium aromaticum, Origanum vulgare, Thymus vulgaris, Hyssopus officinalis , Ocimum basilicum, Myristica fragrans) was determined using the broth microdilution method, which allows reading of the MIC (minimum inhibitory concentration). According to the results obtained, the growth inhibition of Fusarium graminearum was determined by assay for the inhibition of radial growth of the mycelium. RESULTS: The inhibitory effects of thymus, oregano, basil, myristica, hyssop and syzygium essential oil (EO) on mycelial growth of Fusarium graminearum CCM F-683 and CCM 8244 were investigated. The best antifungal activity against the both strains of Fusarium graminearum (37.4%; 40.7%) was demonstrated by Origanum vulgare EO at the concentration 100 µg/mL. Among the four tested oils, three (Syzygium aromaticum, Thymus vulgaris, Origanum vulgare) achieved the best inhibitory effect (100%) at concentrations 500 µg/mL and 1000 µg/mL. CONCLUSIONS: In the protection of plants against pathogenic fungi, essential oils appear to be a suitable substitute for synthetic chemicals.

HPLC-DAD analysis of Hyssopus Cuspidatus Boriss extract and mensuration of its antioxygenation property.

BACKGROUND: Hyssopus cuspidatus Boriss has been used as an important ethnomedicinal plant for long to eliminate phlegm, relieve cough and as well as having antibacterial, antioxygenation, and antitumor activities. In this study, the polyphenol contents, flavonoid contents, free radical scavenging assay and animal antioxygenation property assay of ethanol extract of H. cuspidatus were measured. METHODS: This study determined the total polyphenol and flavonoid contents in H. cuspidatus by UV-VIS. Caffeic, ferulic, and rosmarinic acids were measured using HPLC-DAD. Free radical scavenging assay of H. cuspidatus was studied by colorimetric method. Animal antioxygenation property assay of H. cuspidatus was studied with mice by biochemical assay kits. RESULTS: The total polyphenol and flavonoid contents of H. cuspidatus in 2017, 2018, 2019 were determined and the contents of H. cuspidatus in 2019 was the highest. In addition, rosmarinic acid was the phenolic acid with the highest content in H. cuspidatus. Compared with those of DPPH free radical, hydroxyl free radical, and superoxide anion free radical, the scavenging ability of H. cuspidatus of ABTS free radical was stronger, the average IC50 value was 0.0245 mg/mL. In animal antioxygenation property experiment, the model group was successfully established with decreased activities of SOD, CAT, and GSH-px and increased content of MDA. The ethanol extract of H. cuspidatus increased the activities of SOD, CAT, and GSH-px and reduced the content of MDA. Each group of samples and the ascorbic acid positive control group showed significant differences in the results of free radical scavenging and animal antioxygenation property experiments (P < 0.05). CONCLUSIONS: These results suggest that H. cuspidatus exerts an antioxygenation property, which can be attributed to the contents of total polyphenol and flavonoid. Given its strong antioxygenation property, H. cuspidatus can be used as a new natural antioxidant in food preservation and disease treatment.

Effects of exogenous application of citrulline on prolonged water stress damages in hyssop (Hyssopus officinalis L.): Antioxidant activity, biochemical indices, and essential oils profile.

This study examined the effects of exogenous citrulline (control, 1 and 2 mM) and water availability (100%, 50% and 25% WA) on antioxidant attributes and essential oil constituents of Hyssopus officinalis L. in two successive harvests. Hyssop tolerantly responded to water deficiency by well-promoted antioxidant enzymes (i.e., superoxide dismutase, and catalase), strong DPPH-scavenging activity, and increasing polyphenols; however, the essential oil content was negatively reduced by water stress. External citrulline further increased the activity of antioxidant enzymes. Citrulline application at 2 mM under severe water stress could also improve essential oil (EO) content in the first and second harvests by about 15 and 30%, respectively. Furthermore, under severe drought, citrulline at 2 mM could obtain the highest yield of isopinocamphone (47%) as the main component of EO. The results showed the high potential of this novel applied metabolite agent to be used in a well-fulfilled production of this medicinal plant.

Chemical components of Hyssopus cuspidatus Boriss.: isolation and identification, characterization by HPLC-DAD-ESI-HRMS/MS, antioxidant activity and antimicrobial activity.

Eighteen compounds were isolated from the ethyl acetate fraction of the aerial part of Hyssopus cuspidatus. Their structures were established by analysis of mass and NMR data, as well as comparison with previous published data in the literatures. Among them, ten compounds were found from the Hyssopus genus for the first time, and one compound was isolated from H. cuspidatus for the first time. HPLC-DAD-ESI-HRMS/MS investigations was applied to further obtain the phenolic profiling of the ethyl acetate fraction, and nine derivatives of caffeic acid and ferulic or isoferulic acid were identified. Antioxidant activity against DPPH free radical and antibacterial activity against Candida albicans, Escherichia coli and Staphylococcus aureus were evaluated. The ethyl acetate fraction exhibited weak antioxidant activity and moderate antibacterial activity. The isolated compounds showed weak to potent antioxidant and antibacterial activity. To the best of our knowledge, this is the first report on the antioxidant and antibacterial activity of H. cuspidatus.

Anticancer properties of green-synthesised zinc oxide nanoparticles using Hyssopus officinalis extract on prostate carcinoma cells and its effects on testicular damage and spermatogenesis in Balb/C mice.

The unclear bio-safety issue and potential risk of nanoparticles (NPs) on various organelles can be considered as a major challenge. In the present study, we have assessed the green synthesis of ZnO nanoparticles using Hyssop (Hyssopus officinalis) extract and their effects on PC3 cell line and BALB/c mice model. The cytotoxicity of the ZnO-NPs was assessed on PC3 cell line by MTT test after characterisation. Apoptotic effect of ZnO-NPs was determined by in vitro AO/PI staining. The histopathological assessments and determination of LH and FSH levels carried out as in vivo analysis in BALB/c adult male mice. The expression of major genes involved in spermatogenesis and sperm maturation (Adam3, Prm1, Spata19, Tnp2, Gpx5) were also analysed. The obtained result demonstrated that the IC50 for PC3 cell line treated with green-synthesised ZnO-NPs during 24 and 48 hr was reported 8.07 and 5 µg/ml respectively. Meanwhile, the induced apoptosis was recorded 26.6% ± 0.05, 44% ± 0.12 and 80% ± 0.07 of PC3 cells. The results of gene expression analysis revealed that the increase in the concentration of ZnO-NPs significantly (p < .05) down-regulated the Adam3, Prm1, Spata-19, Tnp2 and Gpx5 genes. The overall results of this research elucidated that ZnO-NPs impaired spermatogenesis, sperm maturation process and sperm motility.

Herbal Extracts Incorporated into Shortbread Cookies: Impact on Color and Fat Quality of the Cookies.

This study aimed at determining the effect of aqueous ethanolic extracts from lemon balm, hyssop and nettle, and butylated hydroxyanisole (BHA) on properties of shortbread cookies. This was achieved by instrumental measurements of color and sensory properties of the cookies directly after baking and by determination of peroxide (PV) and p-anisidine (p-AnV) values, and specific extinction coefficients (K232 and K268 values) for fat extracted from the cookies stored for 3 months at room temperature. Increase of the herbal extracts' concentration from 0.02% to 0.2% in the cookies caused a reduction of L* (the brightness) and a* values (the red coordinate), while b* values (the yellow coordinate) increased when the cookies were enriched with lemon balm and nettle extracts. Among the cookies studied, those prepared with BHA and 0.1 and 0.2% addition of lemon balm extracts were characterized by the highest scores for aroma, taste, and overall acceptability. Incorporation of BHA and 0.02% hyssop extract into the cookies caused a decrease of PV values (the peroxide value) for fat extracted from the cookies after 3 months of their storage compared to a (control) sample without additives and produced the lowest K232 values. Changes in the p-AnV values for the fat samples studied occurred gradually and slowly during the storage and the obtained values were lower compared to the control sample. All of the studied fat samples also showed a higher ability to scavenge DPPH radicals than the control sample. Considering both PV and p-AnV values as indicators of fat oxidation, BHA protected fat extracted from cookies against oxidation better than the herbal extracts used.

Essential Oil Composition of Ruta graveolens L. Fruits and Hyssopus officinalis Subsp. aristatus (Godr.) Nyman Biomass as a Function of Hydrodistillation Time.

The aim of this study was to establish the kinetics regression models for yield and composition of Ruta graveolens fruit and Hyssopus officinalis subsp. aristatus aboveground biomass essential oil (EO), collected at different time intervals during the hydrodistillation process. The hypothesis was that collecting the EO fractions during specific time frames may result in EOs with dissimilar composition that may have differential use by the industry. Furthermore, we calculated the kinetics regression models for the composition of EO, isolated by hydrodistillation in a Clevenger-type apparatus and characterized by GC-MS and GC-FID analyses. The EO yield of R. graveolens fruits was 0.39% (relative area % of GC-FID chromatogram), with major constituents in the Control fraction (0-90 min) being 2-nonanone, 2-undecanone, and 2-undecanol, representing 65% of the total oil. The highest concentration of 2-nonanone (60%) was found in the 30-60 min oil fraction, the concentration of 2-undecanone (35%) was highest in the Control (0-90 min) fraction, and the concentration of eucalyptol (19%) was highest in the 5-10 min fraction. The EO yield of H. officinalis subsp. aristatus dried biomass was 1.12%. The major constituents in the Control fraction (0-90 min) of H. officinalis biomass were eucalyptol, α-pinene, sabinene, ß-pinene, and cis-3-pinanone, representing 86% of the total. Eucalyptol (58%) was the highest in the 0-5 min fraction. The highest ß-pinene (15%) and cis-3-pinanone (20%) contents were found in the 20-40 min fraction. The kinetics regression models that were developed for EO composition of R. graveolens were second-order polynominal, Michaelis-Menten, and Exponential decay, while for EO composition of H. officinalis subsp. aristatus biomass were Exponential decay and Power. The results from this study could benefit the EO industry.

Green synthesis of zinc oxide nanoparticles and evaluation of anti-angiogenesis, anti-inflammatory and cytotoxicity properties.

In this study, zinc oxide nanoparticles (ZnO-NPs) were synthesized using the extract of Hyssops officinalis L. via green method and confirmed by transmission electron microscopy, field emission scanning electron microscopy, X-ray powder diffraction and Fourier transforms infrared spectroscopy techniques. In the in vivo section, the anti-angiogenesis and antiinflammatory properties of the NPs were evaluated by the chorioallantoic membrane (CAM) assay and mouse paw edema test (induced by carrageenan), respectively. In the in vitro section, changes in the expression of angiogenesis genes (VEGF and VEGFR) and inflammatory genes (IL-1B and IL-10) were investigated by real-time quantitative polymerase chain reaction technique. In order to evaluate the cytotoxicity of ZnO-NPs, 3-5, 4-dimethylthiazol-2-yl) -5, 2-tetrazolium bromide (MTT) test was used on MDA-MB231 breast adenocarcinoma cell line. The results of the CAM assay showed that the ZnO-NPs significantly reduced the number and length of blood vessels, as well as the size and weight of the embryos. Evaluation of mouse paw edema showed that the NPs are able to decrease inflammation. Changes in the expression pattern of VEGF and VEGFR genes in MCF7 cells showed that the NPs have inhibitory effect on the expression of both genes. Expression levels of IL-10 and IL-1B genes also increased and decreased, respectively. The MTT test showed that the NP have the ability to decrease breast cancer cells. In conclusion, our results confirm that the ZnO-NPs synthesized by green method have promising anti-cancer properties.

[The expression of apoptosis and autophagy markers in transplantable rat kidney cancer with the administration of flavonoid-containing hedge hyssop ( L.) extract]. / Ékspressiia markerov apoptoza i autofagii v perevitom rake pochki u krys pri vvedenii flavonoidsoderzhashchego ékstrakta avrana lekarstvennogo (L.).

OBJECTIVE: To establish the mechanisms of pathomorphism of transplantable kidney cancer in rats that used flavonoid-containing hedge hyssop (Gratiola officinalis L.) extract in an in vivo experiments. MATERIAL AND METHODS: The experiment was carried out on 30 male Wistar rats with transplantable kidney cancer PA. At 72 hours after tumor inoculation, the rats in the experimental groups received hedge hyssop extract at an oral or intramuscular dose of 110 mg/kg/day for 12 days. A comparison group consisted of the animals with a tumor, but without exposure. The investigators used the immunohistochemical markers of apoptosis (p53, bax, bcl-2, Fas-receptor, Fas-ligand), autophagy (LC3B), proliferation (Ki-67), and angiogenesis (VEGF). During statistical data processing, the Shapiro-Wilk test was used to test the normality of the indicator distribution in the groups. Cramer-Welch's t-test was also employed to compare the groups. RESULTS: Histological examination of tumor tissue under the action of hedge hyssop extract showed the emergence of extensive areas of damage (necrosis and apoptotic bodies). With both routes of hedge hyssop extract administration, there was a sharp decrease in the expression of the proliferation markers Ki-67 and the angiogenesis marker VEGF and a high expression of the apoptosis markers p53, bax, CD95 (Fas/APO-1), and FAS-ligand in tumor cells and its absence in the comparison group. All the described changes were more pronounced with intramuscular administration. The expression of the autophagy marker LC3B increased with the oral administration of hedge hyssop extract and decreased with its intramuscular administration. CONCLUSION: A pronounced pathomorphism of kidney cancer develops due to consumption of hedge hyssop extract. This suppresses the proliferation, angiogenesis, and activation of apoptotic signaling and mitochondrial pathways and blocks protective autophagy. The autophagy marker LC3B can be used as an additional criterion for evaluating the therapeutic pathomorphism of tumors.

JAX2, an ethanol extract of Hyssopus cuspidatus Boriss, can prevent bronchial asthma by inhibiting MAPK/NF-κB inflammatory signaling.

BACKGROUND: Hyssopus cuspidatus Boriss has been used to treat bronchial asthma for many years in Uighur medicine. JAX2, an ethanol extract from this plant, has effectiveness against bronchial asthma. However, the molecular basis for the anti-inflammatory effects of JAX2 remains unclear. PURPOSE: This study aimed to evaluate the mechanism of JAX2 against bronchial asthma. METHODS: We established an asthma model in rats using ovalbumin (OVA), and an inflammatory model in RAW264.7 cells using lipopolysaccharide (LPS) stimulation. To evaluate the anti-inflammatory effects of JAX2, the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-4, IL-6, IL-17, eotaxin and immunoglobulin (Ig)E were measured by enzyme-linked immunosorbent assay (ELISA). Cell viability was investigated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium, inner salt (MTS) assay. Further, nitric oxide (NO) and reactive oxygen species (ROS) were determined using Griess reagent and 2,7-dichlorofluorescein diacetate. The phosphorylation of p-extracellular signal-regulated kinase (ERK), p-Jun-NH2-terminal kinase (JNK), p38 kinases (p38) and p-inhibitor of nuclear factor kappa-B kinase (IKK), and nuclear translocation of p-p65 kinases (p-p65) were determined by immunofluorescence to uncover the effects of JAX2 on the Mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways. RESULTS: After JAX2 administration to rats, Interferon (IFN)-γ concentrations in BALF increased significantly. Further, the concentrations of TNF-α, IL-4, IL-6, IL-17 and eotaxin in BALF, and IgE in serum decreased. JAX2 decreased TNF-α, IL-6 and NO in cell supernatant, and reduced ROS intracellularly. Concurrently, IFN-γ concentrations increased in cell supernatant significantly. In LPS-induced RAW264.7 cells, JAX2 inhibited phosphorylation of p-ERK, p-JNK and p-38 MAPK. The subsequent phosphorylation of p-IKK and nuclear translocation of the p-p65 subunit of NF-κB were also suppressed. CONCLUSION: Based on these findings, we believe that JAX2 has both preventive and treatment effects in bronchial asthma. Furthermore, in the RAW264.7 cell inflammatory model, JAX2 also inhibited NF-κB and MAPK signaling pathways.

Chemical components of Hyssopus seravshanicus: antioxidant activity, activations of melanogenesis and tyrosinase, and quantitative determination by UPLC-DAD.

Hyssopus seravshanicus (Dubj.) Pazij has been used as traditional herb and food due to its wide biological properties. Seventeen known compounds were isolated from the ethyl acetate fraction. Their structures were identified by spectroscopic data and comparison with published data. Among them, 14 ones were identified from H. seravshanicus for the first time. DPPH and ABTS radical scavenging activities for crude ethanol extract (CEE), ethyl acetate fraction (EAF), butanol fraction (BF) and compounds 1, 3, 8, 10, 11 and 13 were performed. CEE, compounds 1, 3, 11 and 13 exhibited potent antioxidant activities. Compound 1 was found to increase the melanin content and tyrosinase activity of B16 melanoma cells. Moreover, the quantitative estimation of compound 1 in the ethyl acetate fraction was carried out by UPLC-DAD and the method was validated. This is the first report on the isolation and bioactivity research on the non-valotile components of H. seravshanicus.

Extraction of Hyssopus officinalis L. essential oil using instant controlled pressure drop process.

To determine the optimum condition for the extraction of Hyssopus officinalis L. essential oil (EO) by instant controlled pressure drop (in French: Détente Instantanée Contrôlée or DIC), the influential parameters of pressure, heating time per cycle, number of cycles, mean particle diameter of the herb, and water concentration in the steam generator feed were evaluated using response surface methodology (RSM). The impact of using an ethanol-water mixture to generate the required vapor in the DIC process on EO extraction was investigated. The optimum condition was found to be at pressure = 1 bar, heating time = 100 s, cycles = 12, particle diameter = 1 mm, and water concentration = 80 v%. The addition of ethanol turned out to be more efficient at higher heating times and cycle numbers. The optimum yield in DIC was highest compared with hydrodistillation (HD), ultrasound-assisted extraction (UAE), and Soxhlet (SOX) methods; their values, expressed as total area percentages per internal standard area percentage, were 39.04, 10.98, 1.58, and 7.71, respectively. Comparison of the four methods also indicated that the DIC-EO was of a superior quality. The analysis of the DIC-treated residual herb in the optimum experiment revealed that unlike the low amount of valuable oxygenated monoterpenes, the availability of sesquiterpenes increased, which proved that DIC performs selectively while extracting more valuable EO components. The DIC-EO and its extract were examined for total phenolic content (TPC) and antioxidant activity using Folin-Ciocalteu and DPPH assay, respectively. Comparison of these results with those obtained from HD demonstrated higher TPC and antioxidant activity of the DIC-EO. Moreover, scanning electron microscopy (SEM) of the raw material and residual herb in the optimum experiment showed significant structure modifications induced by the DIC process.