Bone-Targeted Supramolecular Nanoagonist Assembled by Accurate Ratiometric Herbal-Derived Therapeutics for Osteoporosis Reversal.

Developing novel strategies for defeating osteoporosis has become a world-wide challenge with the aging of the population. In this work, novel supramolecular nanoagonists (NAs), constructed from alkaloids and phenolic acids, emerge as a carrier-free nanotherapy for efficacious osteoporosis treatment. These precision nanoagonists are formed through the self-assembly of berberine (BER) and chlorogenic acid (CGA), utilizing noncovalent electrostatic, π-π, and hydrophobic interactions. This assembly results in a 100% drug loading capacity and stable nanostructure. Furthermore, the resulting weights and proportions of CGA and BER within the NAs are meticulously controlled with strong consistency when the CGA/BER assembly feed ratio is altered from 1:1 to 1:4. As anticipated, our NAs themselves could passively target osteoporotic bone tissues following prolonged blood circulation, modulate Wnt signaling, regulate osteogenic differentiation, and ameliorate bone loss in ovariectomy-induced osteoporotic mice. We hope this work will open a new strategy to design efficient herbal-derived Wnt NAs for dealing with intractable osteoporosis.

Chlorogenic acid intake guidance: Sources, health benefits, and safety.

Chlorogenic acid (CGA) is widely present in plant foods and has attracted much attention due to biological activities such as those which are antioxidant, anti-inflammatory, antibacterial, and antiviral. It plays a role in regulating glucose and lipid metabolism, improving insulin resistance, and reducing the risk of type 2 diabetes and cardiovascular diseases. The estimated dietary intake of CGA is 5 to 1000 mg/d. Based on the data from population intervention studies, daily oral doses of CGA at 13.5mg to 1200 mg can reduce fasting blood glucose (FBG), improve glucose tolerance, enable weight loss /prevent weight gain, and improve blood pressure in hypertensive patients. Daily intake of 200 mg or more may reduce FBG, with a dose-effect relationship in the range 13.5-500 mg/d. Therefore, a specific proposed level (SPL) of CGA to improve FBG could be ≥200 mg/d. Data insufficiency does not allow formulation of a tolerable upper intake level (TUIL) for CGA.

Chlorogenic Acid-Loaded Mesoporous Silica Nanoparticles Modified with Hexa-Histidine Peptides Reduce Skin Allergies by Capturing Nickel.

Nickel-induced contact dermatitis is a severe allergic reaction to objects or environments that contain nickel. Many nanomaterials have been developed to reduce skin allergies by capturing nickel, but few agents are effective and safe. In this work, mesoporous silica nanoparticles (MSN) were synthesized and decorated with hexa-histidine peptides (denoted as MSN-His6), making it a strong nickel chelator. Subsequently, a dietary polyphenol, chlorogenic acid, was loaded into the mesopores of MSN (denoted as MSN-His6@CGA), realizing the potential of its anti-inflammatory properties. In vitro and in vivo experiments revealed that the synthesized MSN-His6@CGA nanoparticles exhibited more stable and stronger chelation, better biocompatibility, and ideal allergy-relieving ability, whether for environmental metal contamination or for allergic contact dermatitis caused by prolonged nickel exposure. Thus, the application of mesoporous silica-based nanoparticles may represent an ideal approach to alleviate skin allergies by capturing nickel, which would benefit people who suffer from metal-induced contact dermatitis.

Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application.

Hepatocellular carcinoma or hepatoma is a primary malignant neoplasm that responsible for 75-90% of all liver cancer in humans. Nanotechnology introduced the dual drug nanodelivery method as one of the initiatives in nanomedicine for cancer therapy. Graphene oxide (GO) loaded with protocatechuic acid (PCA) and chlorogenic acid (CA) have shown some anticancer activities in both passive and active targeting. The physicochemical characterizations for nanocomposites were conducted. Cell cytotoxicity assay and lactate dehydrogenase were conducted to estimate cell cytotoxicity and the severity of cell damage. Next, nanocomposite intracellular drug uptake was analyzed using a transmission electron microscope. The accumulation and localization of fluorescent-labelled nanocomposite in the human hepatocellular carcinoma (HepG2) cells were analyzed using a fluorescent microscope. Subsequently, Annexin V- fluorescein isothiocyanate (FITC)/propidium iodide analysis showed that nanocomposites induced late apoptosis in HepG2 cells. Cell cycle arrest was ascertained at the G2/M phase. There was the depolarization of mitochondrial membrane potential and an upregulation of reactive oxygen species when HepG2 cells were induced by nanocomposites. In conclusion, HepG2 cells treated with a graphene oxide-polyethylene glycol (GOP)-PCA/CA-FA dual drug nanocomposite exhibited significant anticancer activities with less toxicity compared to pristine protocatechuic acid, chlorogenic acid and GOP-PCA/CA nanocomposite, may be due to the utilization of a folic acid-targeting nanodrug delivery system.

Chlorogenic Acid Enhances Doxorubicin-Mediated Cytotoxic Effect in Osteosarcoma Cells.

Despite the recurring outbreak of resistance mechanisms and adverse reactions, doxorubicin (Doxo) still remains the standard-of-care for several cancers, including osteosarcoma (OS). As an appealing source of phytochemical compounds, naturally occurring molecules have extensively been reported to overcome Doxo limitations in preclinical models. Unlike other dietary polyphenols, only few studies recognize chlorogenic acid (CGA) as a potential partner in combination therapy, while, conversely, its anticancer evidence is steadily growing, ultimately in OS. On this basis, herein we examine the cooperating effects between CGA and Doxo in U2OS and MG-63 human OS cells. With respect to Doxo alone, the concomitant administration of CGA further decreased cell viability and growth, promoting cell death potentially via apoptosis induction. Furthermore, a longer-lasting reduction in clonogenic potential deeply supported the CGA ability to improve Doxo efficacy in those cells. Remarkably, CGA treatment ameliorated Doxo-induced cytotoxicity in H9c2 rat cardiomyocyte cells instead. Although inactivation of p44/42 MAPK was detected in response to CGA plus Doxo, PD98059-mediated p44/42 MAPK impairment enhanced the combination outcome in OS cells. These findings firstly propose CGA as a promising chemosensitizer and cardioprotective agent in OS therapy, suggesting the p44/42 MAPK pathway as relevantly involved in CGA-mediated Doxo susceptibility.

Chlorogenic acid ameliorates obesity by preventing energy balance shift in high-fat diet induced obese mice.

BACKGROUND: Chlorogenic acid is a type of phenolic acid found in many plants. Chlorogenic acid has an anti-obesity effect with an unclear mechanism. The present study aimed to investigate the regulatory effect of chlorogenic acid on energy balance in high-fat diet (HFD) induced obese C57BL/6J mice administrated 100 mg kg-1 chlorogenic acid for 13 weeks. RESULTS: The consumption of chlorogenic acid ameliorated HFD induced obesity. Chlorogenic acid did not change the physical activity but significantly decreased food intake and increased body temperature, thermal dissipation and brown adipose tissue activity. Moreover, chlorogenic acid improved glucose tolerance but had a moderate impact on other blood indices. Additionally, chlorogenic acid failed to restore the microbiota change associated with HFD induced obesity, but modified the gut bacterial composition in a unique way. CONCLUSION: Supplementation with chlorogenic acid can improve HFD induced obesity and associated glucose intolerance mainly via regulating food intake and energy expenditure. © 2020 Society of Chemical Industry.

A new method of ultra-performance liquid chromatography/tandem mass spectrometry for determination of chlorogenic acid in human plasma and urine.

RATIONALE: Chlorogenic acid (CA) is well known for its various biological activities. Here, a clinical study was performed in patients with advanced malignant cancer to explore its therapeutic effects. We aimed to develop a method to quantify CA in human plasma and urine to assist the clinical pharmacokinetic study. METHODS: Ultra-performance liquid chromatography (UPLC) coupled with a triple quadruple mass spectrometry was used to separate and detect CA, with puerarin serving as the internal standard. RESULTS: The method presents an excellent linearity ranging from 5 to 2000 ng/mL for plasma analysis and 50 to 20 000 ng/mL for urine analysis. Intra- and inter-day precision and accuracy were both less than 15% for plasma and urine. CONCLUSIONS: These results showed that the novel UPLC method was robust and sensitive, and fulfilled the requirements for a clinical pharmacokinetic study of CA in patients with advanced cancer.

N-(9-Fluorenylmethoxycarbonyl)-L-Phenylalanine/nano-hydroxyapatite hybrid supramolecular hydrogels as drug delivery vehicles with antibacterial property and cytocompatibility.

The intrinsic fragility of hydroxyapatite (HAP) restricts its wider applications for local delivery of antibiotics. The composites formed by integrating HAP with hydrogels can improve the properties of HAP. However, these reported composites not only require tedious preparation and employ organic solvent and toxic reagents, but also hardly have inherent antimicrobial property. In this study, N-(9-Fluorenylmethoxycarbonyl)-L-Phenylalanine/nano-hydroxyapatite (Fmoc-L-Phe/nHAP) hybrid supramolecular hydrogels with antibacterial property and cytocompatibility was prepared by integrating nHAP as reinforcement with Fmoc-L-Phe supramolecular hydrogels. The results showed that nHAP bounds in the chamber of the gel network and adheres to the fiber of Fmoc-L-Phe due to intermolecular interaction, remarkably improving the mechanical strength of Fmoc-L-Phe supramolecular hydrogels. The results of inhibition zone experiment and MTT experiment showed that the Fmoc-L-Phe/nHAP hybrid supramolecular hydrogels possess antimicrobial property and cytocompatibility. In vitro release experiment of chlorogenic acid (CGA) from the hybrid supramolecular hydrogels was performed. The study of the release kinetics indicated that the release behavior of CGA from the hybrid supramolecular hydrogels is following Weibull model and release mechanism involved Fickian diffusion and erosion of the surface of hydrogel matrix. The release of CGA shows a good inhibition effect on S. aureus. The results show that the Fmoc-L-Phe/nHAP hybrid hydrogels with antibacterial property and cytocompatibility have promising applications as drug delivery carrier. Due to the intrinsic fragility of hydroxyapatite (HAP), the properties of HAP could be improved by incorporation into hydrogels. However, these reported composites not only require tedious preparation and employ organic solvent and toxic reagents, but also hardly have inherent antimicrobial property. We prepared N-(9-Fluorenylmethoxycarbonyl)-L-Phenylalanine/nano-hydroxyapatite (Fmoc-L-Phe/nHAP) hybrid supramolecular hydrogels by integrating nHAP as reinforcement with Fmoc-L-Phe supramolecular hydrogels. The results showed that nHAP bounds in the chamber of the gel network and adheres to the fiber of Fmoc-L-Phe due to intermolecular interaction, remarkably improving the mechanical strength of Fmoc-L-Phe supramolecular hydrogels. The results of inhibition zone experiment and MTT experiment showed that the Fmoc-L-Phe/nHAP hybrid supramolecular hydrogels possess antibacterial property and cytocompatibility. In vitro release experiment of chlorogenic acid (CGA) from the hybrid supramolecular hydrogels was performed. The study of the release kinetics indicated that the release behavior of CGA from the hybrid supramolecular hydrogels is following Weibull model and release mechanism involved Fickian diffusion and erosion of the surface of hydrogel matrix. The release of CGA shows a good inhibition effect on S. aureus. The results show that the Fmoc-L-Phe/nHAP hybrid hydrogels with antibacterial property and cytocompatibility have promising applications as drug delivery carrier.

CGA restrains the apoptosis of Aß25-35-induced hippocampal neurons.

Background: Chlorogenic acid (CGA) has anti-oxidant and anti-inflammatory effects, but the study on its role in Alzheimer's disease (AD) models remains rare. Here, the effects of CGA on ß-amyloid protein (Aß)-induced cell models were investigated, aiming to provide a direction for Aß-induced AD.Material and methods: Hippocampal neurons were separated from newborn Sprague-Dawley (SD) rats and identified by immumofluorescence method. Hippocampal neurons were processed with Aß25-35 after pre-treatment CGA. MTT assay was used for detecting viability of treated cells. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and lactate dehydrogenase (LDH) of treated hippocampal neurons were determined by corresponding kits. Flow cytometry analysis assessed the apoptosis and mitochondrial membrane potential (MMP) in hippocampal neurons after treatment. The expressions of proteins related to apoptosis and endoplasmic reticulum stress (ERS) were measured by western blot (WB) analysis.Results: Immumofluorescence method showed that the Aß25-35 induction models were successfully constructed. CGA increased the viability and decreased the apoptosis rate of Aß25-35-induced hippocampal neurons. Decreasing activities of LDH and MDA, and raised contents of SOD and GSH-Px were appeared in Aß25-35-induced cells that pre-treated with CGA. Moreover, CGA also enhanced MMP intensity of hippocampal neurons induced by Aß25-35. In WB analysis, CGA reversed the promoting effect of Aß25-35 on the expressions of proteins related to pro-ERS and pro-apoptosis.Conclusion: CGA restrained the apoptosis of Aß25-35-induced hippocampal neurons via improving the anti-oxidant capacity, mitochondrial injury and ERS state of cells, which may provide a direction for AD.

Green coffee nanoparticles: optimisation, in vitro bioactivity and bio-release property.

Nanoencapsulation by spray drying was performed to protect and preserve antioxidant rich dietary polyphenols from green coffee beans. Nano-encapsulation of green coffee was done using maltodextrin as wall material. The nanoparticles were further characterised by zetasizer, differential scanning colorimetry, X-ray diffraction, In vitro gastric intestinal studies and storage stability. Optimal nanoparticles were obtained at a drying temperature of 125 °C and 2:1 Mwall/Mcore ratio (10% w/w maltodextrin), provided better encapsulation yield (40% w/w) and 70 ± 5% (w/w) encapsulation efficiency with 82.34 nm particle size, -28.8 mV zeta-potential. The In-vitro bioactivity of nanoparticles ensured 80 ± 2% (w/w) of chlorogenic acid availability in a controlled release in the intestine. Storage stability of nanoparticles under varied temperature was remarkably improved compared to non-encapsulated green coffee extract. However, the results indicated that the potential benefits of using maltodextrin coated green coffee nanoparticles for controlled release of Chlorogenic acid and sufficient antioxidative protection during prolonged period.

Nephroprotective effects of chlorogenic acid against sodium arsenite-induced oxidative stress, inflammation, and apoptosis.

BACKGROUND: Chronic exposure to arsenic (As) leads to serious renal disorders. Chlorogenic acid (CGA), a phenolic compound, has several well known physiological benefits, including antioxidant and anti-inflammatory activities. The present study investigated the potential renoprotective effects of CGA on sodium arsenite (NaAsO2 )-induced kidney damage in mice. The mice were randomly allocated into five groups to receive daily treatment with CGA (200 mg kg-1 ), NaAsO2 (5 mg kg-1 ), NaAsO2 + CGA (100 mg kg-1 ), NaAsO2 + CGA (200 mg kg-1 ), or a control for 28 days. RESULTS: In the NaAsO2 -treated group, NaAsO2 induced significant renal dysfunction, oxidative damage, inflammation, and apoptosis, as demonstrated by marked increases in urea and creatinine levels accompanied by a decrease in the kidney index. Considerable increases in malondialdehyde and nitric oxide levels and parallel decreases in various antioxidant markers (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione) levels were also detected in the renal tissues of NaAsO2 -treated mice. NaAsO2 exposure was associated with marked increases in renal inflammatory markers (interleukin-1ß and tumor necrosis factor-α) and apoptosis indicators including Bax and caspase-3 levels contaminant, with a marked decrease in Bcl-2, an anti-apoptotic protein, in the NaAsO2 -treated group compared with the control group. However, pretreatment with CGA substantially mitigated the renal injury and dysfunction associated with NaAsO2 exposure by reducing tissue inflammation and apoptosis and improving the antioxidant status. The CGA pretreatment also alleviated the NaAsO2 -induced histological alterations in renal tissues. CONCLUSION: Taken together, our results suggest the efficacy of CGA in alleviating As-mediated renal tissue damage. © 2020 Society of Chemical Industry.

Chlorogenic acid attenuates kidney fibrosis via antifibrotic action of BMP-7 and HGF.

BACKGROUND: Kidney fibrosis, characterised by tubulointerstitial fibrosis, is a histological landmark of chronic kidney disease. The body attempts to compensate for progressive detrimental process of kidney fibrosis by producing antifibrotic substances, such as bone morphogenetic protein-7 (BMP-7) and hepatocyte growth factor (HGF). Chlorogenic acid is known to have renoprotective and antifibrotic properties. This study aims to evaluate the effect of chlorogenic acid on unilateral ureteral obstruction (UUO)-induced kidney fibrosis mice model. METHODS: This study was a quasi-experimental with posttestonly control group design. Twenty-five adult male Swiss Webster mice were randomly divided into five groups: shamoperated group (SO), UUO-control day-7 (U7), UUO-control day-14 (U14), UUO-chlorogenic acid day-7 (UC7), and UUOchlorogenic acid day 14 (UC14). Myofibroblasts were identified by immunohistochemical staining of alphasmooth muscle actin (α-SMA) while collagen fibers were identified by Sirius Red staining. Both data were presented as area fraction. BMP-7 and HGF mRNA expressions were assessed by reverse transcription PCR (RT-PCR). Data were quantified using ImageJ software. RESULTS: UUO-control groups (U7 and U14) showed higher α- SMA-immunopositive (6.52±1.33, 18.24±1.39 vs. 0.22±0.01; p<0.05) and Sirius Red-positive area fractions (6.61±0.8, 12.98±2.31 vs. 0.62±0.10; p<0.05), lower BMP-7 (1.02±0.47, 1.18±0.65 vs. 2.09±0.87; p<0.05) and HGF mRNA expressions (1.06±0.31, 0.89±0.14 vs. 1.88±0.81; p<0.05) compared to SO group. UUO-chlorogenic acid groups (UC7 and UC14) showed lower α-SMA-immunopositive (1.24±0.37, 4.58±0.61; p<0.05) and Sirius Red-positive area fractions (4.76±1.03, 3.72±0.54; p<0.05), higher BMP-7 (1.84±0.49, 2.19±0.43; p<0.05) and HGF (1.58±0.38; p>0.05, 1.84±0.42; p<0.05) mRNA expressions compared to UUO-control groups. UUOchlorogenic acid groups showed BMP-7 and HGF mRNA expressions that were not significantly different from the SO group. CONCLUSION: Chlorogenic acid administration prevents kidney fibrosis in UUO mice model through modulating antifibrotic pathway.

Coffee consumption and disease correlations.

Coffee is one of the most widely consumed beverages in the world. It has primarily consumed due to its stimulant effect and unique taste since the ancient times. Afterwards, its consumption has been historically associated with a lower risk of some diseases such as type 2 diabetes mellitus, obesity, cardiovascular disease and some type of cancer and thus it has also consumed due to health benefits. It contains many bioactive compounds such as caffeine, chlorogenic acids and diterpenoid alcohols which have so far been associated with many potential health benefits. For example, caffeine reduces risk of developing neurodegenerative disease and chlorogenic acids (CGA) and diterpene alcohols have many health benefits such as antioxidant and chemo-preventive. Coffee also have harmful effects. For example, diterpenoid alcohols increases serum homocysteine and cholesterol levels and thus it has adverse effects on cardiovascular system. Overall, the study that supports the health benefits of coffee is increasing. But, it is thought-provoking that the association with health benefits of coffee consumption and frequency at different levels in each study. For this reason, we aimed to examine the health effect of the coffee and how much consumption is to investigate whether it meets the claimed health benefits.

Changes of porcine gut microbiota in response to dietary chlorogenic acid supplementation.

Chlorogenic acids (CGA), the most abundant natural polyphenol present in human diet and plants, have attracted considerable research interest because of their broad bioactivities including the antimicrobial activity. However, little is known about their influences on intestinal bacterial communities. Here, we described a response in intestinal microbiome to CGA using a porcine model. Twenty-four weaned pigs were allotted to two groups and fed with a basal diet or a basal diet containing 1000 mg/kg CGA. Results showed that CGA significantly increased the length of the small intestine (P < 0.05) and enhanced the activity of diamine oxidase (DAO) and the concentration of MHC-II in the jejunal and ileal mucosa (P < 0.05). Moreover, the acetate concentration in ileum and cecum digesta, and the propionate and butyrate concentrations in the cecum digesta, were significantly elevated by CGA (P < 0.05). Interestingly, CGA significantly increased the total 16S rRNA gene copies and bacterial alpha diversity in the cecum (P < 0.05). The relative abundance of bacteria from phyla Firmicutes and Bacteroidetes was increased in the cecum digesta (P < 0.05), whereas the abundance of bacteria from phylum Protebacteria was decreased by CGA (P < 0.05). Importantly, pigs on CGA-containing diet had higher abundance of Lactobacillus spp., Prevotella spp., Anaerovibrio spp., and Alloprevotella spp. in the cecum (P < 0.05). Not only did our study suggest a synergic response of intestinal barrier function and microbiota to the CGA, but the result will also contribute to understanding of the mechanisms behind the CGA-modulated gut health.

Pharmacokinetics and brain penetration study of chlorogenic acid in rats.

1. The present study is designed to investigate the brain distribution and plasma pharmacokinetics profiles of chlorogenic acid (CGA) after intranasal administration in Charles-Foster rats to evaluate whether the CGA molecules are transported directly via the nose-to-brain path. 2. The CGA is administered intravenously (IV) and intranasally (IN) at the dose of 10 mg/kg. Further, its concentration in the plasma, cerebrospinal fluid (CSF) and the whole brain is analyzed by HPLC-UV method. 3. The study observes that CGA is rapidly absorbed in plasma with tmax of 1 min similar to IV route after IN administration. The peak plasma concentration and AUC0-24 are higher by 3.5 and 4.0 times respectively in IV administration, compared to IN delivery that represents the significant less systemic exposure of CGA in IN route. 4. However, the concentration of CGA in the brain is 4, 6.5, 5.3, 5.2 and 4.5 times higher at 30, 60, 120, 240 and 360 min, respectively in IN administration compared to IV administration. The exposure of CGA in the brain after IN administration (AUCbrain, IN) was significantly greater (4 times) as compared to the exposure of CGA in the brain (AUCbrain, IV) after IV administration reflecting significant brain uptake of CGA through nasal route. Therefore, IN delivery of CGA can be a promising approach for the treatment of stroke and neurodegenerative disorders.

Protective Effects of Kuding Tea (Ilex kudingcha C. J. Tseng) Polyphenols on UVB-Induced Skin Aging in SKH1 Hairless Mice.

In this study, the protective effects of Kuding tea polyphenols (KTPs) on ultraviolet B (UVB)-induced skin injury of SKH1 hairless mice were studied. The ion precipitation method was used for extraction of polyphenols from Kuding tea. High-performance liquid chromatography showed that KTPs contains chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C. SKH1 hairless mice were induced skin aging using 2.0 mW/s intensity of 90 mJ/cm² UV light once a day for seven weeks. The 2.5% and 5% KTPs solution was smeared on 2 cm² of back skin of skin aging mice twice a day. Mouse experiments showed that KTP strongly increased the serum levels of total superoxide dismutase (T-SOD) and catalase (CAT) and reduced those of malondialdehyde, interleukin 6 (IL-6), IL-1ß, and tumor necrosis factor alpha (TNF-α) in mice with UVB-induced skin damage. KTP also increased the levels of type 1 collagen (Col I), hydroxyproline, and hyaluronic acid and reduced those of Col III and hydrogen peroxide in the damaged skin tissues of mice. Pathological observations of tissues stained with H & E, Masson's trichrome, Verhoeff, and toluidine blue showed that KTPs could protect skin cells, collagen, and elastin and decrease the number of mast cells, thus inhibiting skin damage. Quantitative PCR and western blot assays showed that KTP upregulated the mRNA and protein expression of tissue inhibitor of metalloproteinase 1 (TIMP-1), TIMP-2, copper/zinc-SOD, manganese-SOD, CAT, and glutathione peroxidase and downregulated the expression of matrix metalloproteinase 2 (MMP-2) and MMP-9. In addition, the same concentration of KTP had stronger protective effects than vitamin C. The results of this study demonstrate that KTPs have good skin protective effects, as they are able to inhibit UVB-induced skin damage.

Protective effect of chlorogenic acid on lipopolysaccharide-induced inflammatory response in dairy mammary epithelial cells.

Mastitis is a major disease of dairy cattle. Given the recent emergence of antibiotics resistance to mastitis, new intramammary treatments are urgently required. In the present study, we investigated whether lipopolysaccharide (LPS) could induce the increase in the proinflammatory cytokines in bovine mammary epithelial cells (MECs), and whether a natural antimicrobial compound Chlorogenic acid (CGA) could attenuate the inflammatory responses induced by LPS and thus could be a potential therapeutic compound for bovine mastitis. Our results indicated that LPS could induce the expression of proinflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukine (IL)-1ß and IL-6, and the activation of NF-κB p65 and p-p65 in primary bovine MECs. Furthermore, CGA significantly inhibited not only the protein expression of NF-κB p65 and p-p65 but also the mRNA expression of TNF-α, IL-1ß and IL-6 after LPS treatment in primary bovine MECs. These results suggested that CGA had anti-inflammatory role by inhibiting NF-κB activation. In conclusion, CGA could be possibly used as a potential therapeutic compound for bovine mastitis.

Effect of Green Coffee Consumption on Resting Energy Expenditure, Blood Pressure, and Body Temperature in Healthy Women: A Pilot Study.

KEY TEACHING POINTS: • There are several findings, and chlorogenic acid may have antihypertensive effects. • There is a significant correlation between green coffee chlorogenic acid amount and REE. • Intracellular and extracellular liquid amounts decrease following green coffee consumption (215-280 mg caffeine), and these changes are observed in individuals whose usual dietary caffeine intake is relatively both low and high. • The body temperature values of individuals whose usual dietary caffeine intake was in the 50th percentile or greater (relatively high) increased after green coffee consumption.

Chlorogenic acid improves the quality of boar semen subjected to cooled storage at 15°C.

The aim of this study was to investigate the effect of chlorogenic acid (ChA) in boar semen stored at 15°C. Twelve ejaculates were processed into insemination doses at different concentrations of ChA (0.0, 1.5, 3.0, 4.0 and 6.0 mg/ml) or vitamin E (200 µl/ml) as positive control. Semen was analysed after 0, 24 and 72 hr of storage. ChA improved (p < .05) sperm motility, acrosome integrity and mitochondrial activity in all periods of storage. Furthermore, after 24 hr of storage, ChA above 1.5 mg/ml supported the sperm viability until 120 min after reheating (p < .05). Both ChA and vitamin E were similarly efficient in increasing the antioxidant capacity of semen, reducing the malondialdehyde levels before and after 72 hr of storage (p < .05). However, with 72 hr of storage, ChA at 3.0 mg/ml improved the mitochondrial activity over vitamin E (p < .05). In conclusion, results suggest that the concentration of 3.2 mg/ml of ChA is the best for semen stored for up to 24 hr. However, for semen stored for a longer period, 6.0 mg/ml or more should be used.

Influence of 2-Weeks Ingestion of High Chlorogenic Acid Coffee on Mood State, Performance, and Postexercise Inflammation and Oxidative Stress: A Randomized, Placebo-Controlled Trial.

This study measured the influence of 2-weeks ingestion of high chlorogenic acid (CQA) coffee on postexercise inflammation and oxidative stress, with secondary outcomes including performance and mood state. Cyclists (N = 15) were randomized to CQA coffee or placebo (300 ml/day) for 2 weeks, participated in a 50-km cycling time trial, and then crossed over to the opposite condition with a 2-week washout period. Blood samples were collected pre- and postsupplementation, and immediately postexercise. CQA coffee was prepared using the Turkish method with 30 g lightly roasted, highly ground Hambela coffee beans in 300 ml boiling water, and provided 1,066 mg CQA and 474 mg caffeine versus 187 mg CQA and 33 mg caffeine for placebo. Plasma caffeine was higher with CQA coffee versus placebo after 2-weeks (3.3-fold) and postexercise (21.0-fold) (interaction effect, p < .001). Higher ferric reducing ability of plasma (FRAP) levels were measured after exercise with CQA coffee versus placebo (p = .01). No differences between CQA coffee and placebo were found for postexercise increases in plasma IL-6 (p = .74) and hydroxyoctadecadienoic acids (9 + 13 HODEs) (p = .99). Total mood disturbance (TMD) scores were lower with CQA coffee versus placebo (p = .04). 50-km cycling time performance and power did not differ between trials, with heart rate and ventilation higher with CQA coffee, especially after 30 min. In summary, despite more favorable TMD scores with CQA coffee, these data do not support the chronic use of coffee highly concentrated with chlorogenic acids and caffeine in mitigating postexercise inflammation or oxidative stress or improving 50-km cycling performance.