The pro-apoptosis effects of Echinacea purpurea and Cannabis sativa extracts in human lung cancer cells through caspase-dependent pathway

Background: Considering the advantages of using medicinal herbs as supplementary treatments to sensitize conventional anti-cancer drugs, studying functional mechanisms and regulatory effects of Echinacea purpurea (as a non-cannabinoid plant) and Cannabis sativa (as a cannabinoid plant) are timely and required. The potential effects of such herbs on lung cancer cell growth, apoptosis, cell cycle distribution, cellular reactive oxygen species (ROS) level, caspase activity and their cannabinomimetic properties on the CB2 receptor are addressed in the current study. Methods: The cytotoxic effect of both herb extracts on the growth of lung cancer cells (A549) was assessed using the MTT assay. The annexin-V-FITC staining and propidium iodide (PI) staining methods were applied for the detection of apoptosis and cell cycle distribution using flow cytometry. The cellular level of ROS was measured using 7�-dichlorofluorescin diacetate (DCFH-DA) as a fluorescent probe in flow cytometry. The caspase 3 activity was assessed using a colorimetric assay Kit. Results: Echinacea purpurea (EP) root extract induced a considerable decrease in A549 viable cells, showing a time and dose-dependent response. The cell toxicity of EP was accompanied by induction of early apoptosis and cell accumulation at the sub G1 phase of the cell cycle. The elevation of cellular ROS level and caspase 3 activity indicate ROS-induced caspase-dependent apoptosis following the treatment of A549 cells by EP extract. The observed effects of EP extract on A549 growth and death were abrogated following blockage of CB2 using AM630, a specific antagonist of the CB2 receptor. Increasing concentrations of Cannabis sativa (CS) induced A549 cell death in a time-dependent manner, followed by induction of early apoptosis, cell cycle arrest at sub G1 phase, elevation of ROS level, and activation of caspase 3. The CB2 blockage caused attenuation of CS effects on A549 cell death which revealed consistency with the effects of EP extract on A549 cells. Conclusions: The pro-apoptotic effects of EP and CS extracts on A549 cells and their possible regulatory role of CB2 activity might be attributed to metabolites of both herbs. These effects deserve receiving more attention as alternative anti-cancer agents. Graphical abstract: Figure not available: see fulltext. © 2021, The Author(s)

Effects of short-term exposure to naturally occurring thymol concentrations on transmission of a bumble bee parasite

Background: Plants produce antimicrobial phytochemicals that can reduce growth and infectivity of parasites in animals. Pollinator parasites are transmitted between hosts that forage on shared flowers. Floral transmission directly exposes parasites to phytochemicals on floral surfaces and in nectar, both at flowers and, post-ingestion, in the crop. This exposure could directly affect parasite transmission to new hosts. Approach: We combined nectar chemical analyses with field and cell culture experiments to test effects of the floral phytochemical thymol on transmission potential of the trypanosomatid gut parasite Crithidia in Bombus impatiens. First, we measured thymol concentrations in Thymus vulgaris nectar. Second, we tested how addition of thymol to floral nectaries affected parasite transmission to foraging bees. Third, we used cell cultures to determine direct, dose-dependent effects of short term thymol exposure on subsequent in vitro parasite growth. Results: We found 26.1 ppm thymol in Thymus vulgaris nectar, 5-fold higher than previously documented in this species. However, addition of thymol to flowers of parasite-inoculated inflorescences of four plant species did not affect acquisition of Crithidia infection during a foraging bout. Cell culture experiments showed that thymol concentrations needed to reduce subsequent Crithidia growth by 50% (120 ppm) were 4.6-fold higher than the highest detected nectar concentration. Conclusions: Although thymol exposure can influence Crithidia viability, Crithidia are robust to the duration and magnitude of exposure encountered during floral foraging under natural conditions. Our experiments suggest that any effects of thymol alone on Crithidia-host infection dynamics probably reflect indirect, possibly host-mediated, effects of chronic thymol ingestion

Modulation of liver enzymes by an Iranian preparation of Echinacea purpurea

Abstract Hepatitis B, a common infectious disease of liver, is transmitted by blood and body fluids like semen and vaginal fluid that carry hepatitis B virus (HBV). In chronic infection, medical care is required to decrease possibility of cirrhosis and liver cancer. In the present report, the hepatoprotective effect of an Echinacea purpurea preparation (Echiherb ® ) has been described in a patient who suffered from HBV infection. The levels of both enzymes of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) decreased to their normal level after 6 weeks of treatment. Therefore, this report may provide a new perspective for protection of liver in patients with HBV infection along with other diseases which damage liver cells using E. purpurea preparations

Modulation of liver enzymes by an Iranian preparation of Echinacea purpurea

Hepatitis B, a common infectious disease of liver, is transmitted by blood and body fluids like semen and vaginal fluid that carry hepatitis B virus (HBV).  In chronic infection, medical care is required to decrease possibility of cirrhosis and liver cancer. In the present report, the hepatoprotective effect of an Echinacea purpurea preparation (Echiherb®) has been described in a patient who suffered from HBV infection. The levels of both enzymes of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) decreased to their normal level after 6 weeks of treatment. Therefore, this report may provide a new perspective for protection of liver in patients with HBV infection along with other diseases which damage liver cells using E. purpurea preparations

A concern on phthalate pollution of herbal extracts/medicines and detection methods

Esters of phthalates, mainly applied as plasticizer, cause several human health and environment    hazards. Phthalates are widely used in pharmaceutical products, cosmetics, as well as other plastic commercial products, and can penetrate in foods, water dusts, and air leading to ingestion and inhalation exposure followed by skin absorption for human. These compounds cause serious adverse effects on human health like destroying the endocrine system, and consequently developmental alterations and reproductive changes through induction of inflammation and oxidative stress. Some phthalates are able to bio-accumulate in water and have been isolated from aquatic organisms. Mammals and birds may be influenced by these compounds through food chain. Therefore, simple and rapid method for identification and quantification of these compounds is a debate especially for developing countries. Gas Chromatography-Mass Spectroscopy has been successfully employed to determine and measure these compounds in volatile fractions of the plant or the algal materials without more essential chemical reactions. In this article, a rapid review on phthalate toxicity and related analysis methods to detect them in herbal extracts is presented

Monoterpene synthase from Dracocephalum kotschyi and SPME-GC-MS analysis of its aroma profile

Dracocephalum kotschyi (Lamiaceae), as one of the remarkable aromatic plants, widely grows and also is cultivated in various temperate regions of Iran. There are diverse reports about the composition of the oil of this plant representing limonene derivatives as its major compounds. There is no report on cloning of mono- or sesquiterpene synthases from this plant. In the present study, the aroma profile of D. kotschyi has been extracted and analyzed via Headspace Solid-Phase Microextraction technique coupled with Gas Chromatography- Mass Spectroscopy. In order to determine the sequence of the active terpene synthase in this plant, first mRNA was prepared and cloning was performed by 3’ and 5’-RACEs-PCR method, then cDNA was sequenced and finally aligned with other recognized terpene synthases. The results showed that the plant leaves mainly comprised geranial (37.2%), limonene-10-al (28.5%), limonene (20.1%) and 1,1-dimethoxy decane (14.5%). Sequencing the cDNA cloned from this plant revealed the presence of a monoterpene synthase absolutely similar to limonene synthase, responsible in formation of limonene, terpinolene, camphene and some other cyclic monoterpenes in its young leaves

Isolation and characterization of phytochemicals of Johrenia paucijuga (DC.) Bornm.

Background and objectives: The genus Johrenia belongs to Umbelliferae family and contains five species that are endemic to Iran. Johrenia paucijuga grows widely in the north-west, west and center of Iran. So far, there has been no research about phytochemistry of J. paucijuga. In the present study, phytochemicals of the plants have been isolated and their structures have been elucidated. Methods: The aerial parts were dried and cut into small pieces, then extracted with ethyl acetate and methanol using perculator apparatus at room temperature. The methanol extract was extracted again with, petroleum ether and butanol. The separation and isolation process was carried out using column (silica gel and Sephadex LH-20) and thin layer chromatographic (TLC) methods. Structure elucidation of the purified compounds were based on ¹H and ¹³C-NMR data, in comparison with those reported in the previous literatures. Results: The isolated compounds from the ethyl acetate and butanol extracts of J. paucijuga were identified as β-sitosterol, β- stigmasterol and quercetin. Quercetin is a bioactive flavonoid widely used as a health supplement. β- sitosterol and β-stigmasterol are phytosteroles (plants sterols) with chemical structure similar to that of cholesterol and are sometimes used in treating hypercholesterolemia. Conclusion: Regarding the valuable biological properties of the isolated compounds, different biological effects could be expected from the plant

Comparative study of the essential oil and hydrolate composition of Lythrum salicaria L. obtained by hydro-distillation and microwave distillation methods

Essential oils are considered as very complex natural mixtures containing numerous components at quite various concentrations. Lythrum salicaria has not been previously subjected to examination of its volatile oil. The present study was carried out to evaluate the essential oil composition of the flowering aerial parts of the plant by using conventionally hydro-distillation (HD) and microwave assisted hydro-distillation (MAH) methods along with the aromatic water obtained by hydro-distillation. Components of these three samples were recognized by GC/MS. Identification of components resulted in recognition of 14 (98% oil), 32 (98.4%), and 10 (95.5%) compounds for HD, its hydrolate, and MAH samples, respectively. The predominant constituents in the HD oil were bis (2-ethylhexyl) phthalate (29.2%), n-hexadecanoic acid (22%), and pentacosane (9.5%). The major constituents of the hydrolate were elucidated as 2(4H)-benzofuranone-5,6,7,7a-tetrahydro-4,4,7a-trimethyl (6.4%), neryl acetone (6%), pentanoic acid (5.7%), nonanoic acid (5.6%), and octanoic acid (5.5%). Accordingly, bis (2-ethylhexyl) phthalate (43.2%), methyl-cyclopentane (36.5%), and pentacosane (4.7%) were assessed as the main components of MAH oil. Comparing the three samples compositions, it was concluded that the extraction procedure led to variations in quality and quantity of volatiles. Besides, bis (2-ethylhexyl) phthalate, the common plasticizer, was identified as the main pollutant in both HD and MAH oils. The presence of this substance in the oils provided the evidence of contaminations that was absorbed from water into the plant suggesting GC/MS as a suitable method for its detection in plant materials

Anti-inflammatory effect, total polysaccharide, total phenolics content and antioxidant activity of the aqueous extract of three basidiomycetes

Inflammation is a part of the non-specific immune response which occurs in reaction to any type of injury. Medicinal mushrooms have had application in various disorders including cancer, liver injuries, inflammation and diabetes. In the present study, the anti-inflammatory effects of the aqueous extracts of medicinal mushrooms (Fomes fomentarius, Ganoderma applanatum and Trametes hirsuta) were evaluated using carrageenan method. In addition, total polysaccharide, total phenolics contents and the radical scavenging activity of the extracts have also been examined. Mushrooms were extracted with distilled water in 100 °C for 4 hours and then the extracts were freeze dried. Indomethacin was considered as the positive control in the anti-inflammatory evaluation. Polysaccharide contents of F. fomentarius, G. applanatum, and T. hirsuta extracts were assessed as 53.3±0.2, 31.7±0.03, and 19.1±0.6 glucose equivalent µg/100 µgEXT and total phenolic contents of them were successfully revealed as 9.9±0.2, 8.2±0.1, and 8.8±0.2 µgGAE/100 µgEXT, respectively. Furthermore, the IC50 values for F. fomentarius, G. applanatum, and T. hirsuta extracts in DPPH assay, were calculated as 90.9, 108.6, and 908.3 µg/mL, respectively. The results of the experiment showed that the extracts possessed potent anti-inflammatory effect which was comparable to indomethacin