Danhong injection in cardiovascular and cerebrovascular diseases: Pharmacological actions, molecular mechanisms, and therapeutic potential.

Cardiovascular and cerebrovascular diseases are the main cause of mortality worldwide, currently with less than optimum therapeutic options. Danhong injection (DHI) is a medicinal preparation based on two eminent Chinese herbal medicines, Salviae Miltiorrhizae (Dan Shen; family: Lamiaceae) and Flos Carthami (Hong Hua; family: Compositae/Asteraceae). DHI has been mainly used in the clinical therapy of cardiovascular (such as acute coronary syndrome and angina pectoris) and cerebrovascular diseases (such as stroke) in China for many years. The pharmacological properties of DHI include anti-inflammatory, anti-oxidant, anti-coagulatory, hypolipidemic, anti-apoptotic, vasodilatory, and angiogenesis-promoting actions. DHI offers a safe and effective therapeutic agent against cardiovascular and cerebrovascular diseases by modulating multiple disease-relevant signaling pathways and molecular targets. Herein, we provide a comprehensive review of the phytochemistry, therapeutic effects, molecular mechanisms, and adverse reactions of DHI in cardiovascular and cerebrovascular diseases. We also highlight the latest pharmacological advances and therapeutic potential of this promising herb-derived cardiovascular drug preparation.

Differential activation of pregnane X receptor by carnosic acid, carnosol, ursolic acid, and rosmarinic acid.

Pregnane X receptor (PXR) regulates the expression of many genes, including those involved in drug metabolism and transport, and has been linked to various diseases, including inflammatory bowel disease. In the present study, we determined whether carnosic acid and other chemicals in rosemary extract (carnosol, ursolic acid, and rosmarinic acid) are PXR activators. As assessed in dual-luciferase reporter gene assays, carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, activated human PXR (hPXR) and mouse PXR (mPXR), whereas carnosol and ursolic acid, but not carnosic acid or rosmarinic acid, activated rat PXR (rPXR). Dose-response experiments indicated that carnosic acid, carnosol, and ursolic acid activated hPXR with EC50 values of 0.79, 2.22, and 10.77µM, respectively. Carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, transactivated the ligand-binding domain of hPXR and recruited steroid receptor coactivator-1 (SRC-1), SRC-2, and SRC-3 to the ligand-binding domain of hPXR. Carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, increased hPXR target gene expression, as shown by an increase in CYP3A4, UGT1A3, and ABCB1 mRNA expression in LS180 human colon adenocarcinoma cells. Rosmarinic acid did not attenuate the extent of hPXR activation by rifampicin, suggesting it is not an antagonist of hPXR. Overall, carnosic acid, carnosol, and ursolic acid, but not rosmarinic acid, are hPXR agonists, and carnosic acid shows species-dependent activation of hPXR and mPXR, but not rPXR. The findings provide new mechanistic insight on the effects of carnosic acid, carnosol, and ursolic acid on PXR-mediated biological effects.

Effects of the dose of administration, co-antioxidants, food matrix, and digestion-related factors on the in vitro bioaccessibility of rosmarinic acid - A model study.

This study aimed to determine the effect of the administration dose, combinations with co-antioxidants (vitamin C, caffeic acid, chlorogenic acid, catechin, rutin), and different food matrices (cooked and lyophilized hen eggs, chicken breast, soybean seeds, potatoes) on the potential bioaccessibility of rosmarinic acid (RA) in simulated digestion conditions, depending on the digestion stage (gastric and intestinal) and the contribution of physicochemical and biochemical digestion factors. The in vitro bioaccessibility of RA depended on the digestion stage and conditions. The physicochemical factors were mainly responsible for the bioaccessibility of RA applied alone. The higher RA doses improved its bioaccessibility, especially at the intestinal stage of digestion. Furthermore, the addition of vitamin C and protein-rich food matrices resulted in enhanced intestinal bioaccessibility of RA. In the future, the knowledge of factors influencing the bioaccessibility of RA can help enhance its favorable biological effects and therapeutic potential.

Exploring volatile aroma and non-volatile bioactive compounds diversity in wild populations of rosemary (Salvia rosmarinus Schleid.).

Ninety-five rosemary (Salvia rosmarinus Schleid.) genotypes, representing 24 wild populations, collected in different geographical areas in Italy and then cultivated under homogeneous environmental conditions, were characterised for volatile organic compounds (VOCs) and three main non-volatile phenolic diterpenes (carnosic acid, carnosol) and acids (rosmarinic acid). Cluster analysis of chemical data highlighted the occurrence of three main groups of populations: one with high levels of verbenone, α-pinene, bornyl acetate, carnosic acid and carnosol, a second one with relatively high levels of camphor and a third one with high levels of 1,8-cineole, ß-pinene and sesquiterpenes. This clustering was consistent with geographic origin and previous genetic characterisation of the same populations. Correlation analysis suggested that levels of the key antioxidant carnosic acid were associated to levels of the key aroma compound verbenone. The observed diversity may be exploited in breeding programs to develop lines and designing products with improved flavour quality and functional properties.

Insights into the interaction mechanism of acid phosphatase from Lateolabrax japonicus livers and rosmarinic acid using multispectroscopy and molecular docking.

Acid phosphatase (ACP) is a key enzyme that hydrolyzes inosinic acid. The mechanisms underlying the interaction between rosmarinic acid (RA) and ACP and the inhibition of the enzyme were investigated using inhibition kinetics, UV-visible and fluorescence spectroscopy, circular dichroism, and molecular docking. The results showed that RA was a reversible inhibitor of ACP and that the inhibition mechanism was uncompetitive. The ACP fluorescence was quenched by RA, and the quenching mode was static. The interaction of ACP with RA was driven by H bonds and van der Waals forces. The addition of RA increased the α-helix content and decreased the ß-sheet, ß-turn, and random coil contents in ACP, thereby altering the secondary structure of the enzyme. This study enriched our understanding of inhibitory and interaction mechanisms involving ACP and RA.

In vitro bioaccessibility and activity of basil (Ocimum basilicum L.) phytochemicals as affected by cultivar and postharvest preservation method - Convection drying, freezing, and freeze-drying.

The effects of convection drying, freezing, and freeze-drying, on phytochemicals content, in vitro activity and bioaccessibility of sweet basil, cinnamon basil, red rubin basil, and lemon basil were investigated. For evaluation of bioaccessibility, rosmarinic acid content, phenolic content, and antioxidant potential of samples before and after gastric and intestinal steps of digestion were determined. Results showed that the content, activity and bioaccessibility of basil phytochemicals varied depending on the cultivar as well as the applied postharvest preservation. It was found that the frozen and freeze-dried plant materials were characterized by a higher phenolic level and antioxidant activity compared to the convection dried. However, in general, convection drying allows obtaining samples with higher bioaccessibility of phytochemicals compared to the low-temperature processed samples. These findings highlight the need for evaluation of basil cultivars and postharvest preservation methods regarding phytochemicals bioaccessibility, which might help in the classification in terms of pro-health quality.

Involvement of the adenosine A1 receptor in the hypnotic effect of rosmarinic acid.

Insomnia, the most common sleep disorder, is characterized by a longer sleep latency, greater sleep fragmentation, and consequent excessive daytime fatigue. Due to the various side effects of prescribed hypnotics, demand for new drugs is still high. Recent studies have suggested the adenosine receptor (AR) as a potential therapeutic target for insomnia, however, clinically useful hypnotics targeting AR are not yet available. In the present study, we evaluated the hypnotic effect of rosmarinic acid, a phenolic compound widely found in medicinal plants, through pentobarbital-induced sleep test, electroencephalography/electromyography (EEG/EMG), and immunohistochemistry in mice. The underlying mechanisms were assessed by pharmacological approach using 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and SCH5826, antagonists for A1R and A2AR, respectively. Receptor-binding assay and functional agonism were also performed. Our study provides a new evidence that rosmarinic acid has a direct binding activity (Ki = 14.21 ± 0.3 µM) and agonistic activity for A1R. We also found that rosmarinic acid significantly decreased sleep fragmentation and onset latency to NREM sleep, and these effects were abolished by DPCPX. The results from c-Fos immunostaining showed that rosmarinic acid decreased the neuronal activity in wake-promoting brain regions, such as the basal forebrain and the lateral hypothalamus, while increasing the neuronal activity in the ventrolateral preoptic nucleus, a sleep-promoting region; all these effects were significantly inhibited by DPCPX. Taken together, this study suggests that rosmarinic acid possesses novel activity as an A1R agonist and thereby exerts a hypnotic effect, and thus it may serve as a potential therapeutic agent for insomnia through targeting A1R.

A network pharmacology-based study on the quality control markers of antithrombotic herbs: Using Salvia miltiorrhiza - Ligusticum chuanxiong as an example.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza (Danshen, DS), the dried root and rhizome of Salvia miltiorrhiza Bunge and Ligusticum chuanxiong (Chuanxiong, CX), the dried rhizomes of Ligusticum striatum DC are effective in invigorating blood circulation and eliminating stasis which is highly related with cardiovascular disease (CVD). AIM OF STUDY: The identification of activity-based chemical markers is very important, but the complex mechanism of "multi-component, multi-target, and multi-effect" within traditional Chinese medicine (TCM) poses a great challenge to this work. In this study, we combined network pharmacological prediction with experimental validation of the DS and CX to explore an effective method for discovering quality control (QC) of antithrombotic herbs by clarifying the intermediate layer "module/cluster" between the whole complex system and a single component. MATERIALS AND METHODS: Based on structural similarity analysis of compound and the thrombosis network published before, we firstly modularized two layers called chemical cluster (CC) network and functional module (FM) network respectively and linked them into one bilayer modularized compound target (BMCT) network. "Two-step" calculation was applied on identifying the significant compounds as the potential QC markers from CC. The in vitro inhibitory activity of selected QC marker compounds on thrombin was evaluated to partially verify their pharmacological activities. HPLC was used to determine contents. RESULTS: According to the network-based analysis, nine compounds with great importance in the BMCT network were identified as QC markers of DS-CX, including tanshinone I, tanshinone IIA, cryptotanshinone, salvianolic acid B, ferulic acid, salvianolic acid A, rosmarinic acid, chlorogenic acid, and coniferyl ferulate. Enzyme inhibitory test partially verified the activity of tanshinone I and tanshinone IIA. Chemical profiling indicated that the nine marker compounds are the main components in the herbal pair. CONCLUSIONS: This study identified activity-based QC markers of DS-CX herbal pair and provided a new methodology that can be used in the QC of other herbs, herbal pairs, or formulas.

Rosmarinic acid attenuates obesity and obesity-related inflammation in human adipocytes.

Chronic low-grade inflammation is a hallmark of obesity and its related metabolic disorders. At the same time signaling from pro-inflammatory factors such as transforming growth factor beta (TGF-ß) or interleukin 17A (IL-17A) are proposed as crucial for the commitment of fibroblast progenitor cells towards adipogenic differentiation. Modulation of inflammation during adipogenic differentiation is incompletely explored as a potential approach to prevent metabolic disorders. Rosmarinic acid (RA) is a caffeic acid derivative known for its anti-inflammatory effects. Experimental studies of its activity on adipogenic factors or in vivo obesity models are, however, controversial and hence insufficient. Here, we investigated the anti-adipogenic action of RA in human Simpson-Golabi-Behmel syndrome (SGBS) adipocytes. Gene expression levels of key players in adipogenesis and lipid metabolism were assessed. Furthermore, a molecular mechanism of action was proposed. The most prominent effect was found on the translation of C/EBPα, PPARγ and adiponectin, as well as on the modulation of TGF1B and IL17A. Interestingly, involvement of NRF2 signaling was identified upon RA treatment. In summary, our findings indicate that RA prevents inflammation and excessive lipid accumulation in human adipocytes. Data from the molecular analysis demonstrate that RA has potential for treatment of obesity and obesity-related inflammation.

Orange thyme: Phytochemical profiling, in vitro bioactivities of extracts and potential health benefits.

Orange thyme (Thymus fragrantissimus) is becoming widely used in food as a condiment and herbal tea, nevertheless its chemical composition and potential bioactivities are largely unknown. Thus the objective of this work is to obtain a detailed phytochemical profile of T. fragrantissimus by exhaustive ethanolic extraction and by aqueous decoction mimicking its consumption. Extracts showed high content in rosmarinic acid, luteolin-O-hexuronide and eriodictyol-O-hexuronide; these were the main phenolic compounds present in orange thyme accounting for 85% of the total phenolic compounds. Orange thyme extracts presented high scavenging activity against nitric oxide and superoxide radicals. Both extracts presented significant inhibitory effect of tyrosinase activity and moderate anti-acetylcholinesterase activity. Both extracts showed a good in vitro anti-inflammatory activity and a weak anti-proliferative/cytotoxic activity against Caco-2 and HepG2 cell lines supporting its safe use. Orange thyme is a very good source of bioactive compounds with potential use in different food and nutraceutical industries.

Copigmentation evidence of oenin with phenolic compounds: A comparative study of spectrographic, thermodynamic and theoretical data.

The copigmentation effects of polyphenol with different structures vary greatly. Therefore, the aim of this study is to investigate possible interactions in red wine model solutions between oenin and three phenolic compounds: danshensu, caffeic acid and rosmarinic acid. Our results show that the copigmentation of rosmarinic acid is the strongest among the compounds tested. The colourimetric parameters indicate that colour intensity becomes enhanced with increasing concentration of these copigments, leading to darker and more vivid bluish colours. Thermodynamic and quantum chemical investigations are performed to interpret the absorption properties in the visible range. Fluorescence spectroscopy confirms the interaction between caffeic acid and oenin, while FTIR spectroscopic results further suggest a role for hydrogen bonds in the overall process. To our knowledge, this is the first experimentally corroborated direct evidence of hydrogen bonds in copigmentation.

Symphytum officinale L.: Liquid-liquid chromatography isolation of caffeic acid oligomers and evaluation of their influence on pro-inflammatory cytokine release in LPS-stimulated neutrophils.

ETHNOPHARMACOLOGICAL RELEVANCE: Symphytum officinale L. (comfrey, Boraginaceae) has been traditionally used for millennia in joint distortions, myalgia, bone fractures and hematomas. However, key activity-determining constituents and molecular mechanisms underlying its use have not been completely elucidated. AIM OF THE STUDY: The objective of this study was to isolate and identify the major compounds from a hydroethanolic root extract of S. officinale and evaluate their antioxidant potential, alongside their effect on the cytokine production of ex vivo stimulated neutrophils, thus providing scientific support for the traditional use of comfrey root. MATERIAL AND METHODS: Four caffeic acid oligomers were isolated from comfrey roots by liquid-liquid chromatography, their structures being established by MS and NMR analyses. In vitro antioxidant evaluation was performed by DPPH and ABTS assays. The cytotoxicity of isolated compounds was established by flow cytometry. The effect on cytokine release, such as interleukin (IL)-1ß, IL-8 and tumor necrosis factor alpha (TNF-α), in lipopolysaccharide (LPS)-stimulated neutrophils was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The main constituents found in comfrey root were represented by four caffeic acid oligomers, namely globoidnan B (1), rabdosiin (2), rosmarinic acid (3) and globoidnan A (4). Rabdosiin, globoidnans A and B were isolated for the first time from S. officinale. In the in vitro antioxidant tests, compound 2 was the most active, with EC50 values in DPPH and ABTS assays of 29.14 ± 0.43 and 11.13 ± 0.39, respectively. Neutrophils' viability over the tested concentration domain of 12.5-50 µM was not altered. At 50 µM, all compounds significantly inhibited IL-1ß release, with compound 3 (45.60% release vs. LPS stimulated neutrophils) being the most active, followed by compounds 1 (53.85%), 2 (69.89%) and 4 (60.68%). CONCLUSIONS: The four caffeic acid oligomers reported in S. officinale root may contribute to the overall anti-inflammatory activity for which comfrey preparations are used in traditional medicine.

Effect of microwave blanching on antioxidant activity, phenolic compounds and browning behaviour of some fruit peelings.

Fruit peelings are increasingly being used as ingredients for the formulation of functional foods. In this study, the antioxidant activity and browning behaviour of free and bound phenolic fractions of mango, apple, orange and banana peelings microwave-blanched at 720 W for 1, 3 and 5 min were determined. The fractions were equally analyzed for their phenolic content and composition. Generally, increased microwave blanching was accompanied by a significant increase of total phenolics content in free and bound fractions. Increased antioxidant activities at 3 and 5 min of blanching varied with the type of fruit peelings. Browning rates decreased with microwave blanching time except in the case of apple peelings. Epicatechin, ferulic acid, caffeic acid, rosmarinic acid, p-coumaric acid and gallic acid were identified in fruit peelings. Generally, microwave blanching for 3 or 5 min was found to preserve antioxidant activities and reduce the browning rate.

Gynura bicolor DC. (Okinawa spinach): A comprehensive review on nutritional constituents, phytochemical compounds, utilization, health benefits, and toxicological evaluation.

Functional food plants are nature's gift to human beings to help them battle against various ailments for thousands of years. With the global pursuit of improving the quality of life, there is an increasing demand for functional plants in the world's diet. There is a long history of using Gyruna bicolor not only as a vegetable but also as a medicinal herb in East and Southeast Asia. It has been found in G. bicolor that there is a broad spectrum of nutrients and plant bioactives. G. bicolor has been acknowledged by local people as a useful source of medicine for several therapeutic treatments. This paper provides a comprehensive review of G. bicolor from updated research literature. Botanical characteristics, cultivation information, traditional uses, phytochemistry, nutrient values, as well as biological activities are discussed. The storage, processes, proposals for product development, and toxic assessment of G. bicolor are also covered. It is expected that the paper will encourage further research thereby contributing to open avenues for scientific applications of G. bicolor as a valuable plant with multiple health benefits.

Polyphenol composition and biological activity of Thymus citriodorus and Thymus vulgaris: Comparison with endemic Iberian Thymus species.

The polyphenol compositions of Thymus × citriodorus and Thymus vulgaris extracts as obtained by exhaustive hydroethanolic (HE) extraction and aqueous decoction (AD) were compared. In addition, their compositions and bioactivities were compared to those of Thymus pulegioides and Thymus mastichina, grown under the same edaphoclimatic conditions, and Thymus carnosus. Rosmarinic acid was the most abundant polyphenol followed by luteolin-hexuronide, salvianolic acids I and K. Cluster analysis suggests a similarity of the polyphenol composition of T. citriodorus and T. vulgaris. A significant antioxidant activity was observed and correlated with their polyphenol levels. The same being observed for the higher anti-proliferative activity/cytotoxicity of HE extracts on Caco-2 and HepG2 cells as compared to AD extracts. Significant association between the total phenolic compounds with the anti-proliferative activity, for both cell lines, was observed. These results support the importance of salvianolic acids levels in Thymus extracts and their in vitro anti-proliferative/cytotoxic activities.

Pharmacodynamics and pharmacokinetics of Danshen in isoproterenol-induced acute myocardial ischemic injury combined with Honghua.

ETHNOPHARMACOLOGICAL RELEVANCE: Herb pair, the most fundamental and simplest form of herb compatibility, serves as the basic building block of traditional Chinese medicine formulae. The Danshen-Honghua herb pair (DH), composed of Salviae Miltiorrhizae Radix et Rhizoma (Danshen in Chinese) and Carthami Flos (Honghua in Chinese), has remarkable clinical efficacy to cure cardio-cerebrovascular diseases. This study was designed to investigate the pharmacodynamics of DH in comparison with single herbs and pharmacokinetics of DH relative to Danshen in acute myocardial ischemic injury. MATERIALS AND METHODS: Sixty male Wistar rats were divided into control, model and drug treated groups. The acute myocardial ischemia rat model was induced by administering 85 mg/kg/d isoproterenol (ISO) subcutaneously for two consecutive days. For pharmacodynamic study, histopathological and biochemical analysis were performed to assess the anti-myocardial ischemic effects. While for pharmacokinetic study, a UPLC-MS/MS method was developed for determination of nine main active ingredients, namely danshensu, protocatechuic acid, protocatechualdehyde, caffeic acid, lithospermic acid, rosmarinic acid, salvianolic acid B, salvianolic acid A and salvianolic acid C in rat plasma. RESULTS: The histopathological and biochemical analysis revealed that DH exerted enhanced anti-myocardial ischemic effects against the ISO-induced myocardial ischemia compared with single herbs. The pharmacokinetic study indicated that DH could significantly increase the t1/2z of danshensu, Tmax, AUC0-∞ and MRT0-t of protocatechuic acid in comparison with Danshen alone in normal rats, but more importantly elevate systemic exposure level and prolong t1/2z of protocatechualdehyde, caffeic acid, Tmax of danshensu in acute myocardial ischemia rats. CONCLUSIONS: Our findings demonstrated the greater effects of DH after the compatibility in ISO-induced acute myocardial ischemia rats at pharmacodynamic and pharmacokinetic levels and provided valuable information for clinical application of herb pairs.

Ohmic heating polyphenolic extracts from vine pruning residue with enhanced biological activity.

Vine Pruning residue was submitted to conventional heating and ohmic heating (OH) for the extraction of bioactive compounds and analyzed for total phenolic content (TPC), polyphenolic profile, antioxidant activity, antimicrobial activity and anticancer activity. The OH extracts were obtained using Low electric field (496.0 V/cm) or Intermediate electric field - IEF (840.0 V/cm). The tests were performed using 45% (v/v) ethanol-water extraction solution at 80 °C at different extraction times (20-90 min). The extract that stood out among the others concerning anticancer potential was the one obtained by OH when used, IEF, where the TPC was significantly higher than in the other extracts which correlated with higher antioxidant, antimicrobial and anti-proliferative activity on different tumor cell lines (HepG2, MDA-MB-231, MCF-7 and Caco2). Vine pruning OH extracts obtained using green solvents by an eco-friendly procedure were revealed as a source of compounds with relevant antioxidant and anticancer activity.

Identification of acacia honey treated with macroporous adsorption resins using HPLC-ECD and chemometrics.

The adulteration of honey is generally a safety and quality concern for consumers and the industry as a whole. Resin technologies allow harmful substances to enter honey, creating substandard honey, which can enter the market. Thus, it is necessary to identify such illegal products quickly and easily. In this study, HPLC-ECD combined with chemometrics was used to identify raw acacia honey that had been treated with macroporous adsorption resins. The chromatography fingerprints of 46 honey samples were established, and principal component analysis (PCA) and the OPLS-DA identified that differences in some of the chromatographic peaks could be used to distinguish raw from resins-treated raw honeys. 100% correct classification was achieved with test samples, based on the chromatographic peaks identified. These results show that HPLC-ECD, combined with chemometric methods, can identify correctly resins-treated honey and can be applied for the quality control of honey.

Effects of in vitro gastrointestinal digestion and colonic fermentation on a rosemary (Rosmarinus officinalis L) extract rich in rosmarinic acid.

The potential phytochemical losses occurring throughout the sequential steps of in-vitro gastrointestinal digestion and colonic fermentation of a rosemary aqueous extract were investigated. Crude (CE), digested (DE) and fermented (FE) extracts were characterized in terms of their phenolic profile and biological activities. Rosmarinic acid was the phytochemical that underwent the most significate transformation during digestion and fermentation, which amounted to 60% compared to the 26% degradation of the total phenolics. Overall, the simulated digestion step decreased the antioxidant activity estimated by DPPH, ABTS, FRAP, ORAC and TBARS assays. Both CE and DE did not present antiproliferative potential, however, FE exhibited a pronounced cytotoxic activity (GI50 = 116 µg/mL) against HeLa cells. CE and DE showed to be moderate inhibitors of methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus (MSSA), S. aureus, Listeria monocytogenes, whilst the FE acted as a moderate inhibitor of MRSA and MSSA.

Determination of polycyclic aromatic hydrocarbon content in heat-treated meat retailed in Egypt: Health risk assessment, benzo[a]pyrene induced mutagenicity and oxidative stress in human colon (CaCo-2) cells and protection using rosmarinic and ascorbic acids.

This study was undertaken to estimate the concentrations of the formed polycyclic aromatic hydrocarbons (PAHs) in heat-treated (boiled, pan-fried and grilled) meats collected from Egypt. Dietary intakes and cancer risks of PAHs among Egyptian adults were calculated. Benzo[a]pyrene (B[a]P)-induced mutagenicity and oxidative stress in human colon (CaCo-2) cell line and mechanisms behind such effects were also investigated. Finally, protection trials using rosmarinic (RMA) and ascorbic acids (ASA) were carried out. The results indicated formation of PAHs at high levels in the heat-treated meats. Calculated incremental life time cancer risk among Egyptian adults were 7.05179E-07, 7.00604 E-06 and 1.86069 E-05 due to ingestion of boiled, pan-fried and grilled meats, respectively. B[a]P-exposed CaCo-2 cells had high abilities for mutagenicity (490.05 ±â€¯21.37 His + revertants) and production of reactive oxygen species. RMA and ASA protected CaCo-2 cells via reduction of B[a]P-induced mutagenicity and oxidative stress and upregulation of phase II detoxification enzymes and xenobiotic transporters.