Evaluation of Lippia scaberrima Sond. and Aspalathus linearis (Burm.f.) R. Dahlgren extracts on human CYP enzymes and gold nanoparticle synthesis: implications for drug metabolism and cytotoxicity.

BACKGROUND: Metabolism is an important component of the kinetic characteristics of herbal constituents, and it often determines the internal dose and concentration of these effective constituents at the target site. The metabolic profile of plant extracts and pure compounds need to be determined for any possible herb-drug metabolic interactions that might occur. METHODS: Various concentrations of the essential oil of Lippia scaberrima, the ethanolic extract of Lippia scaberrima alone and their combinations with fermented and unfermented Aspalathus linearis extract were used to determine the inhibitory potential on placental, microsomal and recombinant human hepatic Cytochrome P450 enzymes. Furthermore, the study investigated the synthesis and characterization of gold nanoparticles from the ethanolic extract of Lippia scaberrima as a lead sample. Confirmation and characterization of the synthesized gold nanoparticles were conducted through various methods. Additionally, the cytotoxic properties of the ethanolic extract of Lippia scaberrima were compared with the gold nanoparticles synthesized from Lippia scaberrima using gum arabic as a capping agent. RESULTS: All the samples showed varying levels of CYP inhibition. The most potent inhibition took place for CYP2C19 and CYP1B1 with 50% inhibitory concentration (IC50) values of less than 0.05 µg/L for the essential oil tested and IC50-values between 0.05 µg/L-1 µg/L for all the other combinations and extracts tested, respectively. For both CYP1A2 and CYP2D6 the IC50-values for the essential oil, the extracts and combinations were found in the range of 1 - 10 µg/L. The majority of the IC50 values found were higher than 10 µg/L and, therefore, were found to have no inhibition against the CYP enzymes tested. CONCLUSION: Therefore, the essential oil of Lippia scaberrima, the ethanolic extract of Lippia scaberrima alone and their combinations with Aspalathus linearis do not possess any clinically significant CYP interaction potential and may be further investigated for their adjuvant potential for use in the tuberculosis treatment regimen. Furthermore, it was shown that the cytotoxic potential of the Lippia scaberrima gold nanoparticles was reduced by twofold when compared to the ethanolic extract of Lippia scaberrima.

Metabolomic profiling of wild rooibos (Aspalathus linearis) ecotypes and their antioxidant-derived phytopharmaceutical potential.

INTRODUCTION: Aspalathus linearis (commonly known as rooibos) is endemic to the Cape Floristic Region of South Africa and is a popular herbal drink and skin phytotherapeutic ingredient, with health benefits derived primarily from its unique phenolic content. Several, seemingly habitat-specific ecotypes from the Cederberg (Western Cape) and Northern Cape have morphological, ecological, genetic and biochemical differences. OBJECTIVES AND METHODS: Despite the commercial popularity of the cultivated variety, the uncultivated ecotypes are largely understudied. To address gaps in knowledge about the biochemical constituency, ultra-performance liquid chromatography-mass spectrometry analysis of fifteen populations was performed, enabling high-throughput metabolomic fingerprinting of 50% (v/v) methanolic extracts. Antioxidant screening of selected populations was performed via three assays and antimicrobial activity on two microbial species was assessed. The metabolomic results were corroborated with total phenolic and flavonoid screening of the extracts. RESULTS AND DISCUSSION: Site-specific chemical lineages of rooibos ecotypes were confirmed via multivariate data analyses. Important features identified via PLS-DA disclosed higher relative abundances of certain tentative metabolites (e.g., rutin, aspalathin and apiin) present in the Dobbelaarskop, Blomfontein, Welbedacht and Eselbank sites, in comparison to other locations. Several unknown novel metabolites (e.g., m/z 155.0369, 231.0513, 443.1197, 695.2883) are responsible for metabolomic separation of the populations, four of which showed higher amounts of key metabolites and were thus selected for bioactivity analysis. The Welbedacht and Eselbank site 2 populations consistently displayed higher antioxidant activities, with 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities of 679.894 ± 3.427 µmol Trolox/g dry matter and 635.066 ± 5.140 µmol Trolox/g dry matter, respectively, in correlation with a high number of phenolic and flavonoid compounds. The contribution of the individual metabolites to the pharmacological effectiveness of rooibos remains unknown and as such, further structural elucidation and phytopharmacological testing is thus urgently needed.

Rooibos tea waste binary oxide composite: An adsorbent for the removal of nickel ions and an efficient photocatalyst for the degradation of ciprofloxacin.

In this study, rooibos tea waste (RTW) incorporated with a binary oxide (BO; Fe2O3-SnO2) has been reported for the first time as a highly efficient adsorbent material for the elimination of Ni(II) ions. The as-synthesised rooibos tea waste-binary oxide (RWBO) composite adsorbent was characterised using miscellaneous techniques such as FTIR, XRD, SEM, EDX, TGA, BET, and XPS. The RWBO was then tested for the removal of Ni(II) in a batch adsorption experiment. The composite adsorbent showed a great removal efficiency of about 99.75% for Ni(II) ions at 45 °C, 180 min agitation time, pH 7, and dosage of 250 mg. The adsorption process was found to be endothermic and spontaneous. Also, the spent adsorbent [RWBO-Ni(II)] was found to be solar light active with a narrow band gap of 1.4 eV. It was further used as a photocatalyst for the photocatalytic abatement of 10 mg/L ciprofloxacin with an extent of degradation of 83% obtained after 150 min. In addition, the extent of mineralisation of the ciprofloxacin by the spent adsorbent as obtained from the TOC data was found to be 64%. Overall, the RWBO composite adsorbent lends itself as an efficient, eco-friendly and promising material for environmental remediation.

Fermented Rooibos tea (Aspalathus linearis) Ameliorates Sodium Fluoride-Induced Cardiorenal Toxicity, Oxidative Stress, and Inflammation via Modulation of NF-κB/IκB/IκKB Signaling Pathway in Wistar Rats.

High dose of fluoride intake is associated with toxic effects on kidney and cardiac tissues. This study evaluated the potential protective effect of fermented rooibos tea (RTE) on sodium fluoride (NaF)-induced cardiorenal toxicity in rats. Male Wistar rats (n = 56) were randomly allocated into one of seven equal groups: control, NaF (100 mg/kg orally), NaF + RTE (2%, w/v), NaF + RTE (4%, w/v), NaF + lisinopril (10 mg/kg orally), 2% RTE, and 4% RTE. The experiment lasted for 14 days and RTE was administered to the rats as their sole source of drinking fluid. NaF induced cardiorenal toxicity indicated by elevated level of urea, creatinine, LDH, creatinine kinase-MB, and cardiac troponin I in the serum, accompanied by altered histopathology of the kidney and heart. Furthermore, levels of H2O2, malondialdehyde, and NO were elevated, while GSH level was depleted in the kidney and heart due to NaF intoxication. Protein levels of c-reactive protein, TNFα, IL-1B, and NF-κB were increased by NaF in the serum, kidney, and heart. RTE at 2% and 4% (w/v) reversed cardiorenal toxicity, resolved histopathological impairment, attenuated oxidative stress and inhibited formation of pro-inflammatory markers. RTE at both concentrations down-regulates the mRNA expression of NF-κB, and upregulates the mRNA expression of both IκB and IκKB, thus blocking the activation of NF-κB signaling pathway. Taken together, these results clearly suggest that the protective potential of rooibos tea against NaF-induced cardiorenal toxicity, oxidative stress, and inflammation may be associated with the modulation of the NF-κB signaling pathway.

Green rooibos (Aspalathus linearis) promotes gut health: insight into mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: Paralleling the increasing incidence of gastrointestinal disorders world-wide, therapeutic investigations of nutraceuticals to promote gastrointestinal health are gaining popularity. Although anecdotally well-known for its gut health promoting potential, sparse scientific evidence supports this action of Aspalathus linearis (Burm.f.) R. Dahlgren - or rooibos - at the gastrointestinal epithelial level. AIM OF THE STUDY: Traditionally, rooibos is considered to exert antispasmodic, anti-inflammatory, and anti-nociceptive effects in the gut. However, the direct effect on intestinal epithelium is unknown. Thus, to assess the validity of anecdotal claims, two larval zebrafish models were utilized to evaluate effects of rooibos on intestinal health. MATERIALS AND METHODS: Firstly, a larval zebrafish model of gastrointestinal inflammation (2-day TNBS-exposure) was employed. Co-administration of 6α-methylprednisolone served as an internal treatment control. Assessments included live imaging techniques and post-mortem immunofluorescent staining of epithelial tight junction proteins. In addition, whole body H2O2 and prostaglandin E2 assays were performed. Secondly, a gastrointestinal motility assay was performed, with known pro- and anti-kinetic mediators to assess the effect of rooibos to alter functional outcome in vivo. RESULTS: Aqueous and ethanol extracts of green rooibos rescued TNBS-induced reductions in neutral red stained length of larval mid-intestines. Subsequent experiments confirmed the rescue capacity of the aqueous green rooibos extract regarding whole body oxidative and inflammatory status. Concerning tight junction proteins, only the aqueous green rooibos extract - and not prednisolone - normalized both zona occludens-1 and occludin expression levels when compared the TNBS group. In terms of gastrointestinal motility, the aqueous green rooibos extract significantly reduced the extent of gut motility dysregulation achieved by kinetic modulators. CONCLUSIONS: Data indicates the potential of a 2 mg/ml aqueous extract of green rooibos to improve gastrointestinal integrity and functionality in vivo, suggesting beneficial effects of rooibos may already occur at the level of the gut. This provides some evidence to support indigenous knowledge.

Reaction kinetics of aspalathin degradation and flavanone isomer formation in aqueous model solutions: Effect of temperature, pH and metal chelators.

The poor stability of aspalathin in aqueous solutions is a major challenge in delivering a shelf-stable ready-to-drink (RTD) green rooibos iced tea. The kinetics of aspalathin degradation and the formation of eriodictyol glucoside isomers [(S/R)-6-ß-D-glucopyranosyleriodictyol and (S/R)-8-ß-D-glucopyranosyleriodictyol] in aqueous buffers were modeled to understand and predict aspalathin losses during heat processing. The effects of temperature and pH on the rate constants of aspalathin degradation and eriodictyol glucoside isomer formation were determined in a 0.1 M phosphate buffer with 5.7 mM citric acid. The zero-order model best described the reaction kinetics of aspalathin degradation and eriodictyol glucoside isomer formation. Increasing the temperature and pH increased the reaction rate constants. The activation energies of the reactions were much lower at pH 6 than at pH 4, indicating that pH affected the temperature dependence of the reactions. The 8-C-glucosyl eriodictyol derivatives (RE8G and SE8G) formed at much lower rates than the 6-C-glucosyl eriodictyol derivatives (RE6G and SE6G). The metal chelators, citric acid, citrate and EDTA, drastically reduced the reaction rate constants, indicating the catalytic role of metal ions in aspalathin autoxidation. The results of the study could assist manufacturers to improve the shelf life of rooibos RTD beverages by changing the formulation and adjusting heat processing conditions.

Soil water dynamics and biomass production of young rooibos (Aspalathus linearis) plants.

Rooibos (Aspalathus linearis) is endemic to certain regions of the Western- and Northern Cape of South Africa, where it is also commercially grown. Being low-rainfall regions, information on the soil water balance of rooibos is essential, but such data is limited. Consequently, the effect of inorganic fertilisation and soil depth on soil water dynamics in a young rooibos plantation at Nardouwsberg, Western Cape were studied. Soil water content of plots planted to unfertilised and fertilised plants as well as that of bare soil were determined over the duration of the 2016/17 season. All treatments were replicated on shallow and deep soils sites and plant growth was determined at the end of the season. At the end of the study, the profile soil water content and evapotranspiration of the bare and planted plots were similar which prove that fallowing (water harvesting) is not an option in the sandy soils of this region. With the exception of the 20-30 cm root zone of the planted plots at the deep site, the water content decreased to levels below the permanent wilting point in the soil profile during summer. It was concluded that rooibos plants could survive through an adapted root system. A further survival method was proposed, involving moisture moved through evaporation from the deeper soil layers into the drying-front in the ~ 10-30 cm soil layer where a condensation-evaporation cycle enables rooibos to harvest small amounts of water. The highest shoot biomass with the longest taproot resulted from the unfertilised treatment on the deep soil thanks to higher soil water content, whereas the shoot and root biomass of the fertilised treatment at both sites were low due to high P soil concentration. This study revealed that unfertilised plants on deeper soils result in higher rooibos production under drought conditions.

A simple microwave-assisted synthesis of cobalt ferrite nanoparticles and its application for the determination of lead ions in rooibos (Aspalathus linearis) tea.

In this study, a magnetic sorbent assisted dispersive solid phase extraction (DSPE) method was used to preconcentrate lead ions from rooibos tea samples for determination by slotted quartz tube-flame atomic absorption spectrometry (SQT-FAAS). Cobalt ferrite magnetic nanoparticles (CoFe2O4 MNPs) were synthesized by microwave assisted digestion. Limits of detection and quantification were calculated as 5.3 and 17.6 µg/L, respectively, in a linear dynamic range of 20-800 µg/L. The enhancement factor of the developed method was found to be 80-folds when compared to the detection limit of the regular FAAS system. The percent recoveries obtained for rooibos tea samples spiked at different concentrations were in the range of 77 - 125%, with high repeatability as indicated by low standard deviations. The findings of the study demonstrated that the CoFe2O4 MNPs-based extraction method is a straightforward, fast, affordable, safe, and eco-friendly approach to qualifying/quantifying lead with high precision in the selected beverage sample.

Rooibos (Aspalathus linearis) and its constituent quercetin can suppress ovarian cell functions and their response to FSH.

Rooibos (Aspalathus linearis Brum. f) can directly influence female reproduction, but whether rooibos can influence the response of ovarian cells to FSH and whether the rooibos effects are due to the presence of quercetin remain unknown. We compared the influence of rooibos extract and quercetin (both at 10 µg/ml-1) on porcine ovarian granulosa cells cultured with and without FSH (0, 1, 10 or 100 ng/ml-1). The expression of intracellular proliferation (PCNA, cyclin B1) and apoptosis (bax, caspase 3) markers in the cells was detected by immunocytochemistry. The release of progesterone (P), testosterone (T) and estradiol (E) were evaluated with ELISAs. Administration of both rooibos and quercetin reduced the accumulation of proliferation markers and promoted the accumulation of apoptosis markers and the release of T and E. Rooibos stimulated, but quercetin inhibited, P output. Administration of FSH increased the accumulation of proliferation markers, decreased the accumulation of apoptosis markers, promoted the release of P and T, and had a biphasic effect on E output. The addition of both rooibos and quercetin mitigated or prevented the main effects of FSH. The present observations suggest a direct influence of both rooibos and quercetin on basic ovarian functions - proliferation, apoptosis, steroidogenesis and response to FSH. The similarity in the major effects of rooibos and its constituent quercetin indicates that quercetin could be the molecule responsible for the main rooibos effects on the ovary. The potential anti-reproductive effects of rooibos and rooibos constituent quercetin, should be taken into account in animal and human nutrition.

Biocontrol of Fusarium Species Utilizing Indigenous Rooibos and Honeybush Extracts.

Mycotoxins produced by several Fusarium species have a significant effect on reducing maize yield and grain quality and have led to food safety concerns. The antifungal activities of rooibos (Aspalathus linearis) and honeybush (Cyclopia species) tea extracts reduced the growth of plant pathogen Botrytis cinerea, but their efficacy against Fusarium spp. is unknown. In this study, we examined the effects of fermented and unfermented rooibos (A. linearis) and honeybush (Cyclopia subternata) aqueous extracts as well as green tea (Camellia sinensis) against 10 Fusarium species. Conidial viability was assessed by fluorescence microscopy dyes, ATP production was determined using the BacTiter-Glo assay, the mode of action was analyzed by scanning electron microscopy (SEM), and quantification of polyphenols was done using high-performance liquid chromatography with diode array detection (HPLC-DAD). Fermented rooibos extract demonstrated the highest antifungal activity (P < 0.0001) against Fusarium verticillioides MRC 826-E, Fusarium subglutinans MRC 8553, Fusarium proliferatum MRC 8549, and Fusarium globosum MRC 6647, with only 9.53%, 9.26%, 11.0%, and 12.7% ATP production, respectively, followed by antifungal activity of the fermented C. subternata extract against F. subglutinans MRC 8553, F. subglutinans MRC 8554, F. proliferatum MRC 8550, and F. verticillioides MRC 826-E with 3.79%, 6.04%, 6.04%, and 8.40% ATP production, respectively. Extract-treated conidia examined by SEM exhibited disruption of conidial hyphae and collapsed spores. Overall, the fermented rooibos and C. subternata extracts showed higher antifungal activity against the Fusarium species than the unfermented extracts. IMPORTANCE In maize subsistence farming areas in South Africa, daily consumption of maize contaminated by high level of mycotoxins contributes to long-term health effects such as immune deficiency and cancer. Biocontrol methods that are safe and cost-effective are critical to addressing this public health problem. Plant extracts known as biocides or green pesticides are alternatives to chemical pesticides due to their safety and eco-friendly properties. In South Africa, rooibos (Aspalathus linearis) and honeybush (Cyclopia species) contain polyphenols with significant antioxidant and antimicrobial properties. These indigenous herbal teas are widely available and consumed in South Africa and have potential as an innovative approach to reduce mycotoxin levels and, subsequently, human and animal exposure to these toxins. This study evaluates the efficacy of the antifungal activities of several aqueous extracts prepared from fermented and unfermented rooibos (A. linearis), honeybush (Cyclopia subternata), and green tea (Camellia sinensis) on 10 Fusarium strains.

Aspalathus linearis (Rooibos) Targets Adipocytes and Obesity-Associated Inflammation.

Excess weight and obesity are the fifth leading cause of death globally, and sustained efforts from health professionals and researchers are required to mitigate this pandemic-scale problem. Polyphenols and flavonoids found in Aspalathus linearis-a plant widely consumed as Rooibos tea-are increasingly being investigated for their positive effects on various health issues including inflammation. The aim of our study was to examine the effect of Rooibos extract on obesity and the associated low-grade chronic inflammatory state by testing antioxidant activity, cytokine secretions, macrophage polarization and the differentiation of human adipocytes through the development of adipospheroids. Rooibos extract significantly decreased ROS production and the secretion of pro-inflammatory cytokines (IFN-γ, IL-12, IL-2 and IL-17a) in human leukocytes. Additionally, Rooibos extract down-regulated LPS-induced macrophage M1 polarization, shown by a significant decrease in the expression of pro-inflammatory cytokines: TNFα, IL-8, IL-6, IL-1ß and CXCL10. In addition, Rooibos inhibited intracellular lipid accumulation and reduced adipogenesis by decreasing the expression of PPARγ, Ap2 and HSL in adipospheroids. A significant decrease in leptin expression was noted and this, more interestingly, was accompanied by a significant increase in adiponectin expression. Using a co-culture system between macrophages and adipocytes, Rooibos extract significantly decreased the expression of all studied pro-inflammatory cytokines and particularly leptin, and increased adiponectin expression. Thus, adding Rooibos tea to the daily diet is likely to prevent the development of obesity associated with chronic low-level inflammation.

Rooibos tea-in the cross fire of ROS, mitochondrial dysfunction and loss of proteostasis-positioned for healthy aging.

Impaired mitochondrial function and loss of cellular proteostasis control are key hallmarks of aging and are implicated in the development of neurodegenerative diseases. A common denominator is the cell's inability to handle reactive oxygen species (ROS), leading to major downstream oxidative damage that exacerbates neuronal dysfunction. Although we have progressed in understanding the molecular defects associated with neuronal aging, many unanswered questions remain. How much ROS is required to serve cellular function before it becomes detrimental and how does the cell's oxidative status impact mitochondrial function and protein degradation through autophagy? How does ROS regulate autophagy? Aspalathus linearis, also commonly known as rooibos, is an endemic South African plant that is gaining globally acclaim for its antioxidant properties and its role as functional medicinal beverage. In this article we dissect the role of rooibos in the context of the cell's ROS handling capacity, mitochondrial function and autophagy activity. By addressing the dynamic relationship between these critical interconnected systems, and by evaluating the functional properties of rooibos, we unravel its position for preserving cell viability and promoting healthy aging.

Pharmacological activity of Aspalathus linearis extracts: pre-clinical research in view of prospective neuroprotection.

OBJECTIVES: Rooibos tea, a very popular everyday beverage made of Aspalathus linearis plant material and containing multiple polyphenolic compounds, reveals an expectation to positively affect various processes observed in the pathogenesis of neurodegenerative diseases as in the case of consumption of other polyphenol-abundant food products. METHODS: This review is based on available data from pre-clinical in vitro and in vivo studies and presents a broad report on the pharmacological activity of the A. linearis extracts relevant for neurodegenerative diseases. RESULTS: Flavonoids present in herbal infusions are absorbed from gastro-intestinal tract and may affect the central nervous system. The experimental investigations yield the results indicating to supporting role of A. linearis in the prevention of neurodegeneration, primarily owing to anti-oxidative and anti-inflammatory properties, anti-hyperglycaemic and anti-hyperlipidaemic effects as well as favourable impact on neurotransmission with following cognitive and behavioural after-math. DISCUSSION: The multiple pharmacological activities and safety of Aspalathus linearis extracts are commented in the manuscript. The continuous rooibos tea consumption seems to be safe (despite anecdotal liver irritation); however, there is a risk of herbal-drug interactions.

Effect of dietary n-3 fatty acid and rooibos (Aspalathus linearis) supplementation on semen quality, sperm fatty acids and reproductive performance of aged male broiler breeders.

The purpose of this study was to assess the effects of dietary fish oil (FO) and rooibos supplementation on semen quality, fatty acids composition and reproductive performance of aged male broiler breeders. Seventy-two 47-week-old Ross broiler breeder roosters were randomly assigned to a 2 × 3 factorial arrangements to include two FO concentrations (0% and 2%) and 3 rooibos concentrations (0%, 1.5% and 3%) for 13 weeks consecutive. The different diets affected semen parameters significantly (p < 0.05), except for the semen concentration and abnormality of the sperm. The sperm of the FO and 3% rooibos-treated group showed better motility and viability when compared to the other groups (p < 0.05). The susceptibility of semen to lipid peroxidation was increased in roosters fed the rooibos-free diets (p < 0.05), but it was reduced (p < 0.05) when the diet was supplemented with 1.5% and 3% rooibos. In addition, at 64 weeks, the highest concentration of testosterone was observed in the roosters fed a diet that included 2% FO and 3% rooibos (p < 0.05); however, the difference in testosterone levels between Week 52 and Week 64 was not significant (p > 0.05). The fertility rate of collected eggs from the FO and 3% rooibos group was higher (p < 0.05) than that of the other groups at the end of the experiment. In conclusion, dietary inclusion of FO along with rooibos improved seminal quality and reproduction performance in aged roosters.

The toxic contaminants of Aspalathus linearis plant material as well as herb-drug interactions may constitute the health risk factors in daily rooibos tea consumers.

Rooibos tea is brewed using Aspalathus linearis plant material sensitive to environmental contamination. This review covers the safety data from preclinical experiments as well as human studies and delivers a report on its hepatic activity. In vitro tea investigation reveals antioxidative and anti-mutagenic features and ability to modulate microsomal enzymes. In rodent research, it exerts protective or neutral impact on liver functions and morphology, yet several human case reports suggest possible acute hepatic damage. Summarizing rooibos consumption seems to be safe in terms of hepatotoxicity; however, there may be designated a group of consumers with higher risk of liver irritation. The contamination of plant material may contribute to herb-induced liver injury. Due to the impact on CYPs, there is a possible risk of herb-drug interactions affecting bioavailability of some co-administered medicines. Caution should be exercised in patients receiving the treatment with allopathic medicines to avoid untoward alteration of drug plasma concentration.

Aspalathin and Other Rooibos Flavonoids Trapped α-Dicarbonyls and Inhibited Formation of Advanced Glycation End Products In Vitro.

The excessive dietary intake of simple sugars and abnormal metabolism in certain diseases contribute to the increased production of α-dicarbonyls (α-DCs), such as methylglyoxal (MGO) and glyoxal (GO), the main precursors of the formation of advanced glycation end products (AGEs). AGEs play a vital role, for example, in the development of cardiovascular diseases and diabetes. Aspalathus linearis (Burman f.) R. Dahlgren (known as rooibos tea) exhibits a wide range of activities beneficial for cardio-metabolic health. Thus, the present study aims to investigate unfermented and fermented rooibos extracts and their constituents for the ability to trap MGO and GO. The individual compounds identified in extracts were tested for the capability to inhibit AGEs (with MGO or GO as a glycation agent). Ultra-high-performance liquid chromatography coupled with an electrospray ionization mass spectrometer (UHPLC-ESI-MS) was used to investigate α-DCs' trapping capacities. To evaluate the antiglycation activity, fluorescence measurement was used. The extract from the unfermented rooibos showed a higher ability to capture MGO/GO and inhibit AGE formation than did the extract from fermented rooibos, and this effect was attributed to a higher content of dihydrochalcones. The compounds detected in the extracts, such as aspalathin, nothofagin, vitexin, isovitexin, and eriodictyol, as well as structurally related phloretin and phloroglucinol (formed by the biotransformation of certain flavonoids), trapped MGO, and some also trapped GO. AGE formation was inhibited the most by isovitexin. However, it was the high content of aspalathin and its higher efficiency than that of metformin that determined the antiglycation and trapping properties of green rooibos. Therefore, A. linearis, in addition to other health benefits, could potentially be used as an α-DC trapping agent and AGE inhibitor.

The effects of a green Rooibos (Aspalathus linearis) extract on metabolic parameters and adipose tissue biology in rats fed different obesogenic diets.

Current pharmaceutical treatments addressing obesity are plagued by high costs, low efficacy and adverse side effects. Natural extracts are popular alternatives, but evidence for their anti-obesity properties is scant. We assessed the efficacy of a green (minimally-oxidized) Rooibos (Aspalathus linearis) extract (GRT) to ameliorate the effects of obesogenic feeding in rats, by examining body weight, metabolic measures, adipose tissue cellularity and tissue-resident adipose stem cells (ASCs). Furthermore, we performed statistical correlations to explore the relationships and interactions between metabolic and adipose tissue measures. Using an in vivo/ex vivo study design, male Wistar rats were maintained for 17 weeks on one of 3 diets: CON (laboratory chow), OB1 (high-sugar, medium fat) or OB2 (high-fat, high-cholesterol) (n = 24 each). From weeks 11-17, half of the animals in each group received oral GRT supplementation (60 mg per kg body weight daily). Blood and tissue samples were collected, and ASCs from each animal were cultured. Diets OB1 and OB2 induced divergent metabolic profiles compared to CON, but metabolic measures within dietary groups were mostly unaffected by GRT supplementation. Notably, diets OB1 and OB2 uncoupled the positive association between visceral adiposity and insulin resistance, while GRT uncoupled the positive association between elevated serum cholesterol and liver damage. Obesogenic feeding and GRT supplementation induced adipocyte enlargement in vivo, but lipid accumulation in cultured ASCs did not differ between dietary groups. Larger adipocyte size in subcutaneous fat was associated with favourable glucose metabolism measures in all GRT groups. In conclusion, GRT affected the associations between systemic, adipose tissue-level and cellular measures against the background of obesogenic diet-induced metabolic dysregulation.

Rooibos (Aspalathus linearis) alters secretome trace amine profile of probiotic and commensal microbes in vitro.

ETHNOPHARMACOLOGY RELEVANCE: Aspalathus linearis (Burm.f.) R. Dahlgren (rooibos) tea is anecdotally renowned for its calming effect in the context of gastrointestinal discomfort, but little scientific support is available to elucidate potential mechanisms of action. Enhancement of dietary polyphenol content to improve gut health via prebiotic-like modulation of the gut microbiota has gained significant research interest. Given the known high polyphenol content of rooibos, rooibos tea may potentially exert a prebiotic effect in the gut to facilitate an improvement in chronic inflammatory gastrointestinal conditions. AIM OF THE STUDY: This study aimed to determine the prebiotic or health-modulating potential of rooibos tea in terms of its effect on gut microbial growth and secretome trace amine composition, as well as to determine how differential rooibos processing alters this activity. METHODS: Three rooibos preparations (green and fermented leave aqueous extracts, as well as a green leaf ethanol extract) were compared in terms of their phenolic composition (qTOF-LC/MS). Moreover, the effect of rooibos exposure on growth and secretome trace amine levels of probiotic and commensal microbes were assessed (LC/MS). In addition, given the known female bias prevalent for many gastrointestinal disorders, experiments were conducted in the absence and presence of estradiol. RESULTS: Polyphenolic composition of rooibos was drastically reduced by fermentation. Aqueous extracts of both green and fermented rooibos improved microbial growth, although fermented rooibos had the most pronounced effect (p < 0.01). In terms of secretome trace amine profile, both aqueous extracts of rooibos seemed to facilitate increased putrescine secretion (p < 0.0001) and decreased tryptamine production (p < 0.0001). Estradiol seemed to suppress trace amine secretion by bacteria (Lactobacillus plantarum, Lactobacillus reuteri and Enterococcus mundtii) but increased it in yeast (Saccharomyces boulardii). CONCLUSION: Rooibos altered gut probiotic and commensal microbial growth and secretome trace amine profiles in vitro, suggesting it has potential to modulate gut microbial composition and functionality as a prebiotic. Current data suggest that these effects are highly dependent on raw material processing. Finally, rooibos may be able to prevent estradiol-associated alterations in trace amine profile, which may have important implications for patient management in female-predominant gastrointestinal disorders.

Aspalathus linearis suppresses cell survival and proliferation of enzalutamide-resistant prostate cancer cells via inhibition of c-Myc and stability of androgen receptor.

Enzalutamide, a nonsteroidal antiandrogen, significantly prolonged the survival of patients with metastatic castration-resistant prostate cancer (CRPC). However, patients receiving enzalutamide frequently develop drug resistance. Rooibos (Aspalathus linearis) is a shrub-like leguminous fynbos plant endemic to the Cedarberg Mountains area in South Africa. We evaluated the possibility of using a pharmaceutical-grade green rooibos extract (GRT, containing 12.78% aspalathin) to suppress the proliferation and survival of enzalutamide-resistant prostate cancer (PCa) cells. Treatment with GRT dose-dependently suppressed the proliferation, survival, and colony formation of enzalutamide-resistant C4-2 MDV3100r cells and PC-3 cells. Non-cancerous human cells were more resistant to GRT treatment. GRT suppressed the expression of proteins involved in phosphoinositide 3-kinase (PI3K)-Akt signaling, androgen receptor (AR), phospho-AR (Ser81), cyclin-dependent kinase 1 (Cdk1), c-Myc and Bcl-2 but increased the expression of apoptotic proteins. Overexpression of c-Myc antagonized the suppressive effects of GRT, while knockdown of c-Myc increased the sensitivity of PCa cells to GRT treatment. Expression level of c-Myc correlated to resistance of PCa cells to GRT treatment. Additionally, immunofluorescence microscopy demonstrated that GRT reduced the abundance of AR proteins both in nucleus and cytoplasm. Treatment with cycloheximide revealed that GRT reduced the stability of AR. GRT suppressed protein expression of AR and AR's downstream target prostate specific antigen (PSA) in C4-2 MDV3100r cells. Interestingly, we observed that AR proteins accumulate in nucleus and PSA expression is activated in the AR-positive enzalutamide-resistant PCa cells even in the absence of androgen. Our results suggested that GRT treatment suppressed the cell proliferation and survival of enzalutamide-resistant PCa cells via inhibition of c-Myc, induction of apoptosis, as well as the suppression of expression, signaling and stability of AR. GRT is a potential adjuvant therapeutic agent for enzalutamide-resistant PCa.

Optimising the Polyphenolic Content and Antioxidant Activity of Green Rooibos (Aspalathus linearis) Using Beta-Cyclodextrin Assisted Extraction.

Antioxidant activity associated with green rooibos infusions is attributed to the activity of polyphenols, particularly aspalathin and nothofagin. This study aimed to optimise ß-cyclodextrin (ß-CD)-assisted extraction of crude green rooibos (CGRE) via total polyphenolic content (TPC) and antioxidant activity assays. Response surface methodology (RSM) permitted optimisation of ß-CD concentration (0−15 mM), temperature (40−90 °C) and time (15−60 min). Optimal extraction conditions were: 15 mM ß-CD: 40 °C: 60 min with a desirability of 0.985 yielding TPC of 398.25 mg GAE·g−1, metal chelation (MTC) of 93%, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging of 1689.7 µmol TE·g−1, ferric reducing antioxidant power (FRAP) of 2097.53 µmol AAE·g−1 and oxygen radical absorbance capacity (ORAC) of 11,162.82 TE·g−1. Aspalathin, hyperoside and orientin were the major flavonoids, with quercetin, luteolin and chrysoeriol detected in trace quantities. Differences (p < 0.05) between aqueous and ß-CD assisted CGRE was only observed for aspalathin reporting the highest content of 172.25 mg·g−1 of dry matter for extracts produced at optimal extraction conditions. Positive, strong correlations between TPC and antioxidant assays were observed and exhibited regression coefficient (R2) between 0.929−0.978 at p < 0.001. These results demonstrated the capacity of ß-CD in increasing polyphenol content of green rooibos.