Presence Of Chlorogenic Acid Research Articles

Phytochemical, biological, and computational investigations of Ephedra alata Decne. growing in salinity conditions of Arabian Peninsula

Ephedra alata Decne is a medicinal plant widely used in traditional medicine for the management of bronchial asthma and cancer. Phytochemical analysis and biological activities, including antioxidant and anticancer effects, were investigated in the current work as new findings for the plant E. alata, a species growing wildly in the marsh and saline environments of the central area of Saudi Arabia. The Ultra Pressure Liquid Chromatography coupled with Electron spray ionization-Quadropole-Time of flight (UPLC-ESI-Q-TOF) system was used for the phytochemical analysis of the plant constituents. In addition, Polyphenolic profiling including the total phenolic (TPC) and flavonoid (TFC) contents of the plant extracts were measured. Phenolic acids were found at the highest relative percentages among all the identified compounds and were measured at 66.07 mg GAE (Gallic acid equivalent). The UPLC analysis of the E. alata extract indicated the presence of chlorogenic acid, syringic acid, caffeic acid, vanillic acid, rosmarinic acid, umbelliferone, isorhoifolin, and apigenin at the highest relative percentages. Mineral analysis indicated that the microelement content of E. alata was relatively low, except for magnesium (Mg). In vitro antioxidant assays revealed the ability of the plant to scavenge DPPH free radicals, reduced molybdenum ions, and ferrous at levels of 14.63, 19.97, and 27.78 mg Trolox equivalents, respectively. The extract induced transition metal chelation at 31.36 mg EDTA equivalents. The extract induced cytotoxic effects against MDA-231 and A549 cell lines at IC50 levels of 25.31 and 39.81 µg/mL, respectively. The plant extract inhibited the colonization and migration of cancer cells as part of its potential anticancer effects. In addition, major E. alata constituents like isorhoifolin, chlorogenic acid, apigenin, and rosmarinic acid exhibited the lowest binding energy to the CAIX enzyme at − 8.41, − 6.64, − 6.32, and − 6.26 kcal/mol, respectively, compared to the binding energy (− 7.72 kcal/mol) of the co-crystallized ligand (Y0R). The docking results further supported the selection of the CAIX enzyme as a standard predictive therapeutic target, since it exhibited significant binding interactions with the major constituents of the plant.

Screening and identification of reactive metabolic compounds of Cortex Periplocae based on glutathione capture-mass spectrometry.

As a traditional Chinese medicine (TCM), Cortex Periplocae (CP) has a wide range of pharmacological effects, as well as toxic side effects. The main toxic components of it are cardiac glycosides, which tend to cause cardiotoxicity. Currently, it has also been reported in studies to cause hepatotoxicity, but it is not clear whether the hepatotoxicity is related to the toxicity caused by the reactive metabolites. This study aims to investigate the target components of CP that generate reactive metabolic toxicity. The fluorescent probe method was used to detect glutathione (GSH)-trapped reactive metabolites in a co-incubation system of CP extract with rat liver microsomes. Identification of GSH conjugates was performed by LC-MS/MS and that of the possible precursor components that produce reactive metabolites was conducted by UPLC-Q-TOF/MS. Cell viability assays were performed onHepG2 and L02 cells to determine the cytotoxicity of the target components. The findings of our study demonstrate that the extract derived from CP has the ability to generate metabolites that exhaust the intracellular GSH levels, resulting in the formation of GSH conjugates and subsequent cytotoxic effects. Through the utilization of the UPLC-Q-TOF/MS technique, we were able to accurately determine the molecular weight of the precursor compound in CP to be 355.1023. The primary evidence to determining the GSH conjugetes relies on the appearance of characteristic product ions resulting from central neutral loss (CNL) scanning of 129Da and product scanning of m/z 660 in the positive MS/MS spectrum. Through analysis, it was ultimately ascertained that the presence of chlorogenic acid (CGA) and its isomers, namely neochlorogenic acid (NCGA) and cryptochlorogenic acid (CCGA), could lead to the production of GSH conjugates, resulting in cytotoxicity at elevated levels. Taking these findings into consideration, the underlying cause for the potential hepatotoxicity of CP was initially determined.

Enhanced antibacterial and antioxidant activities of chlorogenic acid loaded sweet whey/starch active films for edible food packaging

Starch and whey protein are potential biomaterials for the development of biodegradable and edible food packaging. In this study, chlorogenic acid (CGA), an eco-friendly natural extract, was firstly added to sweet whey/starch matrix as a bioactive compound to develop biodegradable active packaging materials. The results showed that when the ratio between starch and sweet whey was 10:1, water resistance and toughness of sweet whey/starch blend films were improved significantly. After the addition of CGA, the active films showed excellent UV barrier, antibacterial and antioxidant properties. When the addition of CGA reached 3%, UV barrier (at 315 nm), antibacterial (against Escherichia coli) and antioxidant (ABTS method) properties of the active films were increased by 54.6%, 47.5% and 74.5% compared with sweet whey/starch matrix. The presence of CGA could form new hydrogen bonds in the matrix and affect the interactions between sweet whey and starch, which increased swelling rate and toughness of the active films. Preservation experiments of fresh bananas could indicate that the presence of CGA can effectively exert antioxidant effects and inhibit the browning of bananas. As an edible inner coating or film, CGA loaded sweet whey/starch active material has good application potential in the field of food packaging.

In vitro inhibition of 5-α reductase and in vivo suppression of benign prostatic hyperplasia by Physalis angulata ethyl acetate extract

The inhibitory effect against 5-α reductase of the ethyl acetate (EA) extract from Physalis angulata was evaluated in vitro using mouse prostate homogenates, and the suppression of benign prostatic hyperplasia (BPH) was assessed in a mouse model of testosterone-induced BPH. The EA extract exhibited a potentially inhibitory effect on 5-α reductase with an IC50 of 197 μg/ml. In BPH mice, the EA extract at a dose of 12 mg/kg was comparable to finasteride 5 mg/kg in suppressing BPH in terms of reducing absolute enlarged prostate weight (p < 0.05 vs. BPH group) and mitigating the hypertrophy of glandular elements and prostate connective tissue. Identification of chemical ingredients in the EA extract by UPLC-QTOF-MS revealed 37 substances belonging chiefly to flavonoids and physalins. Further quantification of the EA extract by HPLC-PDA methods revealed that chlorogenic acid, and rutin were the main components. Molecular docking studies of chlorogenic acid and rutin on 5-α reductase showed their high affinity to the enzyme with binding energies of −9.3 and − 9.2 kcal/mol, respectively compared with finasteride (− 10.3 kcal/mol). Additionally, chlorogenic acid inhibited 5-α reductase with an IC50 of 12.07 µM while rutin did not. The presence of chlorogenic acid in the EA extract may explain the inhibitory effects of the EA extract on 5-α reductase, and thus the suppression of BPH.

Cytotoxic and Antitumoral Effects of Methanolic Extracts of Avocado Fruit Mesocarp in Colorectal Cancer Cell Line HT29.

Colorectal cancer is a widespread neoplasia with high ratios of chemoresistance. Phytochemicals in plant-based extracts could be useful to treat colorectal cancer, and/or reduce chemoresistance. Methanolic extract of avocado mesocarp (MEAM) has demonstrated antitumoral properties, depending on the fruit ripening stage (RS). The aim of this study was to analyze the effects of methanolic extracts of "Hass" avocado fruit at different RS on cytotoxicity, antioxidative, anti-inflammatory, anti-invasive, cell cycle, and epithelial-mesenchymal transition inhibition in colorectal adenocarcinoma cell line HT29. The MEAM showed an increasing concentration of total phenolic compounds as the RS progressed, which was correlated with antioxidant capacity measured by the Ferric Reducing Antioxidant Power assay but not with the 2.2-diphenyl-1-picrylhydrazyl assay. The specific phenolic compounds of MEAM were determined by high-performance liquid chromatography, and it was found that concentrations of epicatechin decreased while concentrations of chlorogenic acid increased as the RS progressed. The HT29 cell line was treated with MEAM for 48 h, and all MEAM had a cytotoxic effect, reported by MTT assay, nevertheless, the strongest effect was associated with the presence of chlorogenic acid. MEAM induced apoptosis and cell cycle arrest in phase G0/G1, reported by flow cytometry. Moreover, MEAM inhibited cell migration evidenced by the wound healing assay. On the other hand, MEAM significantly reduced expression of mRNA of tumor necrosis factor-alpha and cyclooxygenase 2. These effects comprise important inhibition of some hallmarks of cancer. This, in turn, may provide interesting guidelines for developing antitumoral intervention agents.

Improving the functionality of pea protein with laccase-catalyzed crosslinking mediated by chlorogenic acid

The influences of laccase-catalyzed crosslinking on the structural, emulsifying and gelling properties of pea protein with chlorogenic acid were intensively probed. Molecular weight analysis illustrated the formation of pea protein aggregates by laccase-induced polymerization in the presence of chlorogenic acid and the increase of incubation time facilitated the aggregation. Particle size of pea protein-laccase-chlorogenic acid progressively enhanced increasing incubation time. Laccase-induced polymerization possessed remarkable impacts on the secondary and tertiary structure of pea protein, confirmed by circular dichroism, and fluorescence spectroscopy. Surface hydrophobicity of pea protein appreciably enhanced with laccase-induced crosslinking due to the exposure of interior hydrophobic amino acid residues. Emulsifying activity, emulsifying capacity, gel strength, and water-holding capacity of pea protein can be considerably enhanced with laccase-catalyzed crosslinking with chlorogenic acid, suggesting excellent functionalities for pea protein were accomplished after being modified by laccase with chlorogenic acid. The obtained information will widen pea protein’s application in food systems.

Effects of Extraction Methods on Phenolic Content, Antioxidant and Antiplatelet Activities of Tomato Pomace Extracts

Aqueous and ethanolic extracts of tomato pomace were examined with the aim of optimizing the extraction process of compounds with cardioprotective activity. Once the results of the ORAC response variables, total polyphenols, °Brix, and antiplatelet activity of the extracts were obtained, a multivariate statistical analysis was performed using the Statgraphics Centurion XIX software. This analysis showed that the most relevant positive effects in the inhibition of platelet aggregation were 83 ± 2% when using the agonist TRAP-6, when the working conditions were the type of tomato pomace conditioning (drum-drying process at 115 °C), phase ratio (1/8), type of solvent (ethanol 20%), and type of extraction (ultrasound-assisted solid-liquid extraction). The extracts with the best results were microencapsulated and characterized by HPLC. The presence of chlorogenic acid (0.729 mg/mg of dry sample) was found, a compound that has a potential cardioprotective effect documented in various studies, in addition to rutin (2.747 mg/mg of dry sample) and quercetin (0.255 mg/mg of dry sample). These results show that the extraction efficiency of compounds with cardioprotective activity depends largely on the polarity of the solvent, thus playing an important role in the antioxidant capacity of the extracts of tomato pomace.

Effect of Cu2+ and Al3+ on the interaction of chlorogenic acid and caffeic acid with serum albumin

Despite the phenolic acids' health benefits, their interactions with proteins are still unclear. In this study, the interactions of Bovine Serum Albumin (BSA) with chlorogenic acid (CHA), caffeic acid (CA), and their Al3+, Cu2+ complexes were studied by using circular dichroism (CD) spectroscopy, fluorescence spectroscopy, and UV/Vis spectroscopy. It was found that esterification of carboxyl group of CA with quinic acid increased the binding affinities for BSA. After chelating with Cu2+ and Al3+, both CHA and CA exhibited high binding affinities for BSA. CHA could form CHA-Cu2 and CHA-Al2 complex with Cu2+ and Al3+. The result of CD spectroscopy demonstrated that the binding of CHA and Al3+ with BSA contributed to the folding of BSA secondary structure. In addition, with the presence of CHA, binding with Al3+ could also induce changes in BSA conformation. The binding sites of both CHA and CA were closed to Trp213.

Valorization of Raw Coffee Beans (Coffea arabica and Coffea canephora) through Solvent Development and Extraction of Bioactive Compounds

Raw coffee beans are seen as valuable sources of bioactive compounds, such as alkaloids and chlorogenic acids. In this study, an efficient and eco-friendly method for the simultaneous extraction of caffeine, trigonelline, and chlorogenic acids from raw beans of two coffee species was developed, using green solvents and an unconventional method of extraction (assisted by ultrafast rotary disintegrator/homogenizer (UT-AE)). The experimental extraction conditions were optimized according to a completely randomized design (CRD), considering the following variables: solvent type (four deep eutectic solvents (DESs), water, and aqueous choline chloride solution (50 wt.%); temperature (25, 45, and 65 °C); and extraction technique (solid-liquid extraction with agitated heating and assisted by ultrafast rotary disintegrator/homogenizer). The extract obtained with choline chloride solution (50 wt.%) exhibited high total phenolic compounds and the highest antioxidant capacity. An analysis using high-performance liquid chromatography with a diode array detector (HPLC-DAD) indicated the presence of chlorogenic acids, caffeine, and trigonelline in all the extracts, in different amounts. The results obtained by the analysis of phenolic compounds and HPLC indicated that the aqueous solution of choline chloride (50% wt.%) was the most suitable solvent for the extraction of chlorogenic acids, while the water-based extracts showed high values of caffeine and trigonelline. DESs, in turn, seems to promote a protective effect on the antioxidant activity of biomolecules.

The Application of Ultrasonic Waves and Microwaves to Improve Antihyperglycaemic and Antimicrobial Activities of Marrubium vulgare Extracts.

Having scarce information about ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE) of white horehound (Marrubium vulgare L.), the idea has emerged to determine the optimal process parameters for the maximization of polyphenols and to compare the efficiency of these green extraction technologies. The optimal UAE parameters are temperature of 73.6 °C, extraction time of 40 min and ultrasound power of 30.3 W/L, while the optimal MAE parameters are 63.8% ethanol, extraction time of 15 min and microwave power of 422 W. Extract obtained at optimal UAE parameters shows the highest antihyperglycemic activity (α-amylase inhibition: 50.63% and α-glucosidase inhibition: 48.67%), which can potentially be explained by the presence of chlorogenic acid and quercetin, which were not identified in the macerates. The most sensitive bacterial strain to optimal ultrasonic extract is Bacillus cereus, whereas the most sensitive fungal strain is Saccharomyces cerevisiae.

The insecticidal effect of the botanical insecticide chlorogenic acid on Mythimna separata (Walker) is related to changes in MsCYP450 gene expression.

The oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae) can feed on the leaves of many crops, resulting in vast areas of damage and severe losses. Therefore, this insect has become a significant agricultural pest in north Asia. In this study, we fed 3rd instar larvae with artificial diets containing different concentrations of chlorogenic acid and found a significant lethal effect and the mortality increased with increasing chlorogenic acid concentration. Next, we measured the sublethal effect of chlorogenic acid at LC20 on the growth and development of M. separata larvae. The durations of the 4th and 5th instar were longer than those of the control group (prolonged by 0.8 and 0.6 days, respectively), and the 6th instar was shorter (by 1.1 days). The total survival rate, pupation rate, eclosion rate, sex ratio, and oviposition amount in the LC20 chlorogenic acid-treated group were significantly lower than those in the control group. Furthermore, transcriptome analysis of 3rd instar larvae fed various concentrations of chlorogenic acid revealed that several MsCYP450 genes were significantly up-regulated, and this finding was further validated by qRT-PCR. In addition, various concentrations of chlorogenic acid and different treatment times significantly affected the enzyme activity of CYP450 in 3rd instar larvae. Importantly, dietary ingestion of dsMsCYP450 significantly reduced the mRNA level of MsCYP450 genes and increased mortality in the presence of chlorogenic acid. Our results revealed that MsCYP6B6, MsCYP321A7, and MsCYP6B7-like play an essential role in the detoxification of chlorogenic acid by M. separata. This study provides evidence of control effect by botanical insecticide chlorogenic acid on M. separata, and potential detoxification mechanism mediated by P450 of botanical insecticide in arthropods.

A dispersive liquid-liquid microextraction followed by reverse-phase high-performance liquid chromatography for QuEChERS determination of chlorogenic acid.

The presence of chlorogenic acid (CGA) not only imparts a characteristic flavour to foods but also makes them useful against chronic diseases and metabolic syndromes, especially diabetes mellitus and asthma. The present study was designed to develop a quick, easy, cheap, effective, rugged, and safe (QuEChERS) and pragmatic method to analyse CGA in plant-based products by applying dispersive liquid-liquid microextraction (DLLME) followed by reverse-phase high-performance liquid chromatography under a diode array detector (RP-HPLC-DAD). The DLLME was carried out using Triton X-100 as a dispersant and ethanol as an extraction solvent, while method development and validation activities were performed on a Shimadzu 10-AT HPLC-DAD system equipped with C-18 columns as stationary phases. The well-resolved peak for the standard CGA was observed at 2.92 min for the mobile phase comprising 0.1% aqueous formic acid-ethanol (22:78 v/v) of pH 3.0 programmed under an isocratic flow rate of 1.0 mL/min. The developed method was found to be linear (R2 = 0.9976) over 1-500 μg/mL of CGA concentration at 328 nm. Moreover, the assay was found to be repeatable with RSD ≤ 5.0, and the limit of detection (LOD) and limit of quantification (LOQ) were 0.0281 and 0.0853 μg/mL of CGA, respectively. The DLLME offered an overall recovery rate between 97.60% and 99.54% at an acceptable level of reproducibility [relative standard deviation (RSD) ≤ 5.0]. The developed assay was found to be a QuEChERS and pragmatic choice for the high-throughput analysis of CGA in plant-based products/foods. Finally, the analysis revealed the presence of an ample level of CGA in guava fruit in addition to coffee beans and black tea.

Molecular Docking and Efficacy of Aloe vera Gel Based on Chitosan Nanoparticles against Helicobacter pylori and Its Antioxidant and Anti-Inflammatory Activities.

The medicinal administration of Aloe vera gel has become promising in pharmaceutical and cosmetic applications particularly with the development of the nanotechnology concept. Nowadays, effective H. pylori treatment is a global problem; therefore, the development of natural products with nanopolymers such as chitosan nanoparticles (CSNPs) could represent a novel strategy for the treatment of gastric infection of H. pylori. HPLC analysis of A. vera gel indicated the presence of chlorogenic acid as the main constituent (1637.09 µg/mL) with other compounds pyrocatechol (1637.09 µg/mL), catechin (1552.92 µg/mL), naringenin (528.78 µg/mL), rutin (194.39 µg/mL), quercetin (295.25 µg/mL), and cinnamic acid (37.50 µg/mL). CSNPs and A. vera gel incorporated with CSNPs were examined via TEM, indicating mean sizes of 83.46 nm and 36.54 nm, respectively. FTIR spectra showed various and different functional groups in CSNPs, A. vera gel, and A. vera gel incorporated with CSNPs. Two strains of H. pylori were inhibited using A. vera gel with inhibition zones of 16 and 16.5 mm, while A. vera gel incorporated with CSNPs exhibited the highest inhibition zones of 28 and 30 nm with resistant and sensitive strains, respectively. The minimal inhibitory concentration (MIC) was 15.62 and 3.9 µg/mL, while the minimal bactericidal concentration (MBC) was 15.60 and 7.8 µg/mL with MBC/MIC 1 and 2 indexes using A. vera gel and A. vera gel incorporated with CSNPs, respectively, against the resistance strain. DPPH Scavenging (%) of the antioxidant activity exhibited an IC50 of 138.82 μg/mL using A.vera gel extract, and 81.7 μg/mL when A.vera gel was incorporated with CSNPs. A.vera gel incorporated with CSNPs enhanced the hemolysis inhibition (%) compared to using A.vera gel alone. Molecular docking studies through the interaction of chlorogenic acid and pyrocatechol as the main components of A. vera gel and CSNPs with the crystal structure of the H. pylori (4HI0) protein supported the results of anti-H. pylori activity.

Chlorogenic Acid Prevents UVA-Induced Skin Photoaging through Regulating Collagen Metabolism and Apoptosis in Human Dermal Fibroblasts.

Skin aging is categorized as chronological aging and photo-aging that affected by intrinsic and extrinsic factors. The present study aimed to investigate the anti-aging ability and its underlying mechanism of chlorogenic acid (CGA) on human dermal fibroblasts (HDFs). In this study, CGA specifically up-regulated collagen I (Col1) mRNA and protein expressions and increased the collagen secretion in the supernatant of HDFs without affecting the cell viability, the latter was also demonstrated in BioMAP HDF3CGF system. Under ultraviolet A (UVA)-induced photoaging, CGA regulated collagen metabolism by increasing Col1 expression and decreasing matrix metalloproteinase 1 (MMP1) and MMP3 levels in UVA-irradiated HDFs. The activation of transforming growth factor-β (TGF-β)-mediated Smad2/3 molecules, which is crucial in Col1 synthesis, was suppressed by UVA irradiation and but enhanced at the presence of CGA. In addition, CGA reduced the accumulation of UVA-induced reactive oxygen species (ROS), attenuated the DNA damage and promoted cell repair, resulting in reducing the apoptosis of UVA-irradiated HDFs. In conclusion, our study, for the first time, demonstrate that CGA has protective effects during skin photoaging, especially triggered by UVA-irradiation, and provide rationales for further investigation of CGA being used to prevent or treat skin aging.

Phytochemical analysis, antioxidant, cytotoxic, and antimicrobial activities of golden chamomile (Matricaria aurea (Loefl.) Schultz Bip).

Matricaria aurea (Loefl.) Schultz Bip. (Asteraceae), known as golden chamomile, has been traditionally used for the treatment of various diseases. In this study, total phenolic, flavonoid, and tannin contents of total extract and different fractions of this plant were determined. The antioxidant, cytotoxic, and antimicrobial activities were also evaluated. Moreover, the phenolic profiles of selected fractions were determined by HPLC and LC-MS/MS analysis. Results demonstrated total phenolic contents of 37.8-57.2mg GAE/g and total flavonoid contents of 3.0-111.2mg QE/g. The ethyl acetate and methanol fractions (EF and MF) had the highest concentrations of phenolic, tannin, and flavonoid compounds. In both DPPH radical scavenging assay and phosphomolybdenum reduction assay, EF showed the best antioxidant activity, followed by MF. EF and MF indicated also the best antibacterial activities against Bacillus subtilis (MIC 1.56 and 12.5mgml-1) and Staphylococcus aureus (MIC 0.78 and 12.5mgml-1). Hexane fraction (HF) had no antibacterial effect. None of the samples had antifungal effect. MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay revealed for EF and HF the highest antiproliferative activities (IC50 values ranged from 111.8 to 294.6μgml-1). The presence of chlorogenic acid, ferulic acid, and luteolin-7-O-glucoside in MF, and p-coumaric acid in EF was confirmed and quantified.

Chemical compositions of Sideritis albiflora Hub. – Mor.

Background and Aims: Sideritis species (Lamiaceae) which are quite widespread in the Mediterranean region and represented by 46 species and 53 taxa in Turkey, are often used as a antirheumatic, anti-inflammatory, antimicrobial and diuretic remedies. Methods: In this study, the infusion (INF), essential oil (EO) and methanolic extract (ME) of chemical compounds prepared from aerial parts of Sideritis albiflora Hub.-Mor. (endemic) were investigated. Results: The presence of chlorogenic acid, verbascoside, forsythoside, apigenin glucoside, and isoscutellarein derivatives in the infusion and methanol extract of the plant was determined by LC-MS / MS analysis. In addition, the main constituents of the essential oil were found as germacrene D (23.5%), ß-caryophyllene (13.6%) and caryophylene oxide (8.0%) by GC / MS and GC / FID simultaneously. Conclusion: In this study, the essential oil, the methanolic extract and the infusion of S. albiflora species were prepared and their chemical compositions elucidated. It was determined that the essential oil chemical compound composition is rich in terms of the sesquiterpene group. Infusions of aerial parts of some Sideritis species have traditionally been used in the treatment of many diseases. Accordingly, the fact that the phenolic compounds of S. albiflora infusion have not been studied before increases the importance of this study. In addition, the therapeutic effect was shown to be related to major S. albiflora compounds, and the correlation between in vitro activity and ethnobotanical use was evaluated. The effective phenolic compounds contained in S. albiflora are thought to support the traditional uses of the plant.

Study on Chlorogenic Acid Inhibiting the Proliferation and Invasion of Fibroblast-Like Synoviocytes in Rheumatoid Arthritis Model

This paper explored Chlorogenic acid regulating the biological behavior of RA FLSs and studied the functional role of microRNAs in it. In vivo experiment: Female DBA/1 J mice were used for model establishment and grouping. HE staining was employed. The damage of ankle cartilage was analyzed in each group of mice. The levels of serum cytokines TNF-α and IL-β were measured by ELISA. In vitro experiment: The cells were counterstained with Hoechst 33342, Transwell was used to detect cell invasion. Western blotting was used to detect the expression of Akt protein. The Akt expression plasmid and miR-23b mimic were co-transfected into RA FLSs, and the luciferase activity was measured using a dual-luciferase detection system. In vivo experiments found that Chlorogenic acid can significantly reduce arthritis index and inhibit TNF-α and IL-β levels. In vitro experiments found that TNF-α-induced proliferation of RA FLSs was significantly inhibited by Chlorogenic acid. Transwell invasion test showed that TNF-α-induced cell invasion was attenuated at the presence of Chlorogenic acid, which significantly inhibited Akt protein expression and phosphorylation. The expression of miR-23b in Chlorogenic acid-treated RA-FLSs increased, and silencing miR-23b enhanced the inhibitory effect of RA FLSs on Chlorogenic acid induction. Chlorogenic acid has potential anti-rheumatoid arthritis activity. Its inhibition of RA FLSs proliferation and invasion is related to the induction of miR-23b and the down-regulation of Akt expression.

Phytochemical screening and antioxidant and antimicrobial activities of crude extracts of different filamentous fungi.

Five filamentous fungal strains that grew in different whey-based media under submerged fermentation were investigated for antioxidant properties and phytochemicals. Phytochemical analysis revealed the presence of alkaloids, tannin, flavonoids, glycosides, phenols, saponins, and terpenes in the crude intra- and extracellular ethyl acetate extracts of different strains. All fungal extracts exhibited effective antioxidant activities in terms of TPC, TFC, DPPH, FRAP, ABTS, reducing power, and metal chelating capacity. The activities of intracellular extracts were higher than the extracellular metabolites. Fermentation media with sugar and salt supplementation significantly influenced antioxidant production. Aspergillus niger in glucose-supplemented whey medium was found to exhibit the highest antioxidant properties. The antimicrobial activity of A. niger and Penicillium expansum extracts by microtiter plate assay showed a promising result against some pathogenic bacterial strains. Chromatographic analysis of the fungal extracts revealed the presence of chlorogenic acid, trans-cinnamic acid, ferulic acid quercetin, myricetin, kaempferol, and catechin which are known for their antioxidant properties. Accumulation of nutrients in fungal biomass under constraint environment produces secondary metabolites which has demonstrated efficacy towards alleviation of several degenerative diseases. The antioxidative enriched phytochemicals present in these five different fungal strains will provide a breakthrough in the utilisation of whey as inexpensive source of substrate for the growth of these fungi. Moreover, phytochemicals could be utilized as therapeutic agents in a cost-effective and environmentally friendly manner.

Pharmacognostical, phytochemical and physiochemical studies of Bridelia tomentosa Bl.

To provide a scientific basis for identification of Bridelia tomentosa Bl., the current study was carried out for pharmacognostical and preliminary phytochemical analysis. The roots, stems and leaves were used for character identification, microscopic identification and physicochemical parameters analysis which mainly included the commonly used methods, like ash value, UV absorption spectrum and thin layer chromatography (TLC) etc. Pharmacognostic studies revealed presence of non-glandular hairs in leaf midrib transverse section, calcium oxalate cluster crystals and calcium oxalate crystals in stem transverse section, fibers in root transverse section, cork cells, fibers, non-glandular trichomes, pitted vessels, calcium oxalate crystals and calcium oxalate crystal clusters in powder microscopy. Phytochemical investigation showed the presence of carbohydrates, tannins and volatile oils from water and petroleum ether soluble extract, while flavones, phytosterol and triterpene were detected from ethanol extract and alkaloids from acid-water soluble extract. The presence of chlorogenic acid in Bridelia tomentosa extract with specific Rf values was detected using TLC. The findings confirmed that Bridelia tomentosa clearly has significant pharmacognostical characters, which would be valuable for providing a dependable foundation for authenticity of plant materials.

Protective Effect of Chlorogenic Acid on Human Sperm: In Vitro Studies and Frozen-Thawed Protocol.

The study evaluated the chlorogenic acid (CGA) antioxidant potential on oxidative stress (OS) induced in vitro in human spermatozoa and during cryopreservation procedure. Swim-up selected spermatozoa were treated with 100 µM CGA, 100 µM H2O2 to induce lipid peroxidation (LPO), and with both compounds and the effects on mitochondrial membrane potential (MMP) by JC-1, DNA integrity by acridine orange (AO), and sperm ultrastructure by transmission electron microscopy (TEM), were evaluated. CGA antioxidant activity was assessed by measuring malondialdehyde (MDA) and F2-isoprostanes (F2-IsoPs) in the media. The CGA protective activity and the immunolocalization of Phospho-AMPKα (Thr172) were explored in frozen-thawed sperm. CGA was not toxic for sperm motility, DNA integrity and MMP. The increase in MDA (p < 0.05) and F2-IsoPs (p < 0.001), DNA damage (p < 0.01) and low MMP (p < 0.01) levels after H2O2 treatment were reduced in presence of CGA as well as the percentage of broken plasma membranes (p < 0.01) and altered acrosomes (p < 0.01) detected by TEM. Treated frozen-thawed spermatozoa showed increased sperm motility (p < 0.01), DNA integrity (p < 0.01), MMP (p < 0.01), reduced MDA (p < 0.01) and increased sperm percentage with Phospho-AMPKα labelling in the head (p < 0.001). CGA can be used to supplement culture media during semen handling and cryopreservation where OS is exacerbated.