Chemoselective Preparation of New Families of Phenolic-Organoselenium Hybrids-A Biological Assessment.

Being aware of the enormous biological potential of organoselenium and polyphenolic compounds, we have accomplished the preparation of novel hybrids, combining both pharmacophores in order to obtain new antioxidant and antiproliferative agents. Three different families have been accessed in a straightforward and chemoselective fashion: carbohydrate-containing N-acylisoselenoureas, N-arylisoselenocarbamates and N-arylselenocarbamates. The nature of the organoselenium framework, number and position of phenolic hydroxyl groups and substituents on the aromatic scaffolds afforded valuable structure-activity relationships for the biological assays accomplished: antioxidant properties (antiradical activity, DNA-protective effects, Glutathione peroxidase (GPx) mimicry) and antiproliferative activity. Regarding the antioxidant activity, selenocarbamates 24-27 behaved as excellent mimetics of GPx in the substoichiometric elimination of H2O2 as a Reactive Oxygen Species (ROS) model. Isoselenocarbamates and particularly their selenocarbamate isomers exhibited potent antiproliferative activity against non-small lung cell lines (A549, SW1573) in the low micromolar range, with similar potency to that shown by the chemotherapeutic agent cisplatin (cis-diaminodichloroplatin, CDDP) and occasionally with more potency than etoposide (VP-16).

Decreased Human Platelet Activation and Mouse Pulmonary Thrombosis by Rutaecarpine and Comparison of the Relative Effectiveness with BAY11-7082: Crucial Signals of p38-NF-κB.

Platelets play a critical role in arterial thrombosis. Rutaecarpine (RUT) was purified from Tetradium ruticarpum, a well-known Chinese medicine. This study examined the relative activity of RUT with NF-κB inhibitors in human platelets. BAY11-7082 (an inhibitor of IκB kinase [IKK]), Ro106-9920 (an inhibitor of proteasomes), and RUT concentration-dependently (1-6 µM) inhibited platelet aggregation and P-selectin expression. RUT was found to have a similar effect to that of BAY11-7082; however, it exhibits more effectiveness than Ro106-9920. RUT suppresses the NF-κB pathway as it inhibits IKK, IκBα, and p65 phosphorylation and reverses IκBα degradation in activated platelets. This study also investigated the role of p38 and NF-κB in cell signaling events and found that SB203580 (an inhibitor of p38) markedly reduced p38, IKK, and p65 phosphorylation and reversed IκBα degradation as well as p65 activation in a confocal microscope, whereas BAY11-7082 had no effects in p38 phosphorylation. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay shows that RUT and BAY11-7082 did not exhibit free radical scavenging activity. In the in vivo study, compared with BAY11-7082, RUT more effectively reduced mortality in adenosine diphosphate (ADP)-induced acute pulmonary thromboembolism without affecting the bleeding time. In conclusion, a distinctive pathway of p38-mediated NF-κB activation may involve RUT-mediated antiplatelet activation, and RUT could act as a strong prophylactic or therapeutic drug for cardiovascular diseases.

Targeting Production of Reactive Oxygen Species as an Anticancer Strategy.

Cancer remains the second leading cause of death worldwide. Research is currently focused on finding novel anticancer therapies and elucidating their mechanisms of action. Cellular redox balance is a promising target for new therapies, as cancer cells already have elevated levels of oxidizing agents due to hypermetabolism and genetic instability. Although free radicals are actively involved in vital cellular signaling pathways, they have also been implicated in certain diseases, including cancer. The aim of this review was to highlight the involvement of oxidative stress in the mechanism of action of anticancer agents. The difference in cellular redox balance between normal and cancer cells is discussed as a potential anticancer target, along with various examples of approved or experimental drugs that may alter the redox state. These drugs are presented in relation to their pro-oxidant or antioxidant mechanisms, with the consequent goal of underscoring the importance of such mechanisms in the overall efficacy of anticancer drugs.

Anthocyanins from Pomegranate (Punica granatum L.) and Their Role in Antioxidant Capacities in Vitro.

As phytochemicals, anthocyanins are not only responsible for the diverse colors in nature, but are associated with broad-spectrum health-promoting effects for human beings. Pomegranate is abundant in anthocyanins which possess high antioxidant capacities. However, the pomegranate anthocyanins profile and their contributions to antioxidant capacities are not fully depicted. The purpose of this article is to review anthocyanins from pomegranate as important antioxidants. Total anthocyanin content (TAC) and six major components vary greatly with intrinsic and extrinsic factors. In pomegranate, anthocyanins mainly acted as primary antioxidants, while their action as secondary antioxidants were not conclusive. The antioxidant potentials of anthocyanins were significantly affected by factors especially chemical structure and detection assays in vitro. The current knowledge may provide insights into potential applications for pomegranate anthocyanins based on their antioxidant activities.

Purification and Identification of Novel Antioxidant Peptides from Enzymatically Hydrolysed Samia ricini Pupae.

The emergence of excessive free radicals leads to the destruction of various systems within the body. These free radicals also affect nutritional values, color, taste, and emit an odor akin to rancid food. Most food industries use synthetic antioxidants, such as BHT (butylated hydroxytoluene) or BHA (butylated hydroxy anisole). However, high doses of these can be harmful to our health. Therefore, an antioxidant compounds, such as bioactive peptides from edible animals or plants, have emerged to be a very promising alternative as they reduce potential side effects. This study focused on the purification and identification of antioxidant peptides from protein hydrolysates of wild silkworm pupae (Samia ricini). Antioxidant peptides were purified from the hydrolysate by ultrafiltration and RP-HPLC. The results showed that protein hydrolysate from S. ricini pupae by trypsin with a molecular weight lower than 3 kDa and highly hydrophobic property, exhibited strong DPPH radical scavenging activity and chelating activity. Further identification of peptides from the fraction with the highest antioxidant activity was carried out using LC-MS/MS. Three novel peptides, i.e., Met-Ley-Ile-Ile-Ile-Met-Arg, Leu-Asn-Lys-Asp-Leu-Met-Arg, and Glu-Asn-Ile-Ile-Leu-Phe-Arg, were identified. The results of this study indicated that the protein hydrolysate from S. ricini pupae possessed potent biological activity, and the novel antioxidant peptides could be utilized to develop health-related antioxidants in food industry.

The mechanism and prevention of mitochondrial injury after exercise.

With the development of society, physical activity has come to be an effective means by which people pursue good health to improve the quality of life. However, with the increase of intensity and the passage of time, exercise injury has become a hazard that can no longer be ignored. It is imperative to find effective ways to inhibit or reduce the negative effects of exercise. Mitochondria are important organelles involved in exercise and play an important role in exercise injury and prevention. Studies have found that exercise preconditioning and increased mitochondrial nutrition can effectively decrease mitochondrial damage after exercise. Against this background, some of the newest developments in this important field are reviewed here. The results discussed indicate that exercise preconditioning and supplement mitochondrial nutrition need to be increased to prevent exercise-related injuries.

Synthesis, Characterization and Biological Evaluation of Indole-Pyrazole Amalgamated α-Cyano Substituted Chalcones.

BACKGROUND: Indole and pyrazole constitute a major class of biologically active scaffolds. The amalgamation of two or more pharmacophores would generate novel molecular templates that are likely to unveil remarkable biological properties. OBJECTIVE: An efficient and high yielding synthesis of indole-pyrazole integrated α-cyano substituted chalcones and their in vitro anti-breast cancer and antioxidant evaluation. METHODS: The synthesis of a series of indole-pyrazole amalgamated α-cyano substituted chalcones (6a-o) was achieved by reacting substituted 3-cyanoacetyl indole 2 with substituted pyrazole aldehyde 5 in the presence of piperidine. All the newly synthesized compounds have been characterized by IR, 1H NMR and HRMS spectroscopy. RESULTS: Anti-breast cancer evaluation of the synthesized compounds in vitro against MCF-7 cell line revealed high anti-breast cancer activities. Amongst the compounds screened 6f, 6g, 6h, 6c, 6d, 6e, 6i and 6k unveiled excellent activity against breast carcinoma (GI50 <0.1µM) as good as adriamycin (GI50 <0.1µM). The compounds were also screened against the normal Vero monkey cell line and the results demonstrated more selectivity against MCF-7. On the other hand, compounds 6b, 6c, 6d, 6h and 6i have shown moderate DPPH and NO radical scavenging activity. CONCLUSION: Most of the synthesized compounds exhibited significant antitumor activities. These results further support its safety margin by studying the activity on normal Vero monkey cell line. These results acclaim the possible use of these compounds for the design and development of potent anti-breast cancer agents.

Cascade Reactions by Nitric Oxide and Hydrogen Radical for Anti-Hypoxia Photodynamic Therapy Using an Activatable Photosensitizer.

Organelle-targeted activatable photosensitizers are attractive to improve the specificity and controllability of photodynamic therapy (PDT), however, they suffer from a big problem in the photoactivity under both normoxia and hypoxia due to the limited diversity of phototoxic species (mainly reactive oxygen species). Herein, by effectively photocaging a π-conjugated donor-acceptor (D-A) structure with an N-nitrosamine substituent, we established a unimolecular glutathione and light coactivatable photosensitizer, which achieved its high performance PDT effect by targeting mitochondria through both type I and type II (dual type) reactions as well as secondary radicals-participating reactions. Of peculiar interest, hydrogen radical (H•) was detected by electron spin resonance technique. The generation pathway of H• via reduction of proton and its role in type I reaction were discussed. We demonstrated that the synergistic effect of multiple reactive species originated from tandem cascade reactions comprising reduction of O2 by H• to form O2•-/HO2• and downstream reaction of O2•- with •NO to yield ONOO-. With a relatively large two-photon absorption cross section for photoexcitation in the near-infrared region (166 ± 22 GM at 800 nm) and fluorogenic property, the new photosensitizing system is very promising for broad biomedical applications, particularly low-light dose PDT, in both normoxic and hypoxic environments.

Antimutagenic, Antiproliferative and Antioxidant Properties of Sea Grape Leaf Extract Fractions (Coccoloba uvifera L.).

BACKGROUND: Cancer is a disease characterized by the invasion and uncontrolled growth of cells. One of the best ways to minimize the harmful effects of mutagens is through the use of natural antimutagens. In this regard, the search for new antimutagens that act in the chemoprevention could represent a promising field in this area. OBJECTIVE: In this study biological potential of 11 fractions from Coccoloba uvifera L. leaf hexane extract was evaluated by several in vitro tests. METHODS: Leaves were lyophilized and hexane extraction was performed. The extract was fractionated by column chromatography with hexane, ethyl acetate, and methanol. The antimutagenic (Ames test), antiproliferative (MTT test), and antioxidant capacity (DPPH, ABTS, and ferrous ion chelation) of the fractions were evaluated. RESULTS: Fractions 4, 6, 8, and 9 have antimutagenic activity (against sodium azide in strain TA100), fraction 11 showed antiproliferative capacity (IC50 of 24 ± 9 µg/mL in cells of HCT 116). The fractions with the highest activity were analyzed by HPLC-MS and lupeol, acacetin, and ß-sitosterol were identified. CONCLUSION: This study demonstrates, for the first time, the bioactivity of C. uvifera leaf as a new source of High Biological Value Compounds (HBVC), which can be of interest to the food and pharmaceutical industries.

The diterpene Manool extracted from Salvia tingitana lowers free radical production in retinal rod outer segments by inhibiting the extramitochondrial F1 Fo ATP synthase.

Uncontrolled oxidative stress production, especially in the outer retina is one of the causes of retinal degenerations. Mitochondria are considered the principal source of oxidative stress. However, a Reactive Oxygen Intermediates (ROI) production in the retinal photoreceptor layer seems to depend also on the expression of an extramitochondrial oxidative phosphorylation (OxPhos) machinery in the rod outer segments (OS). In fact, OS conduct aerobic metabolism, producing ATP through oxygen consumption, although it is devoid of mitochondria. As diterpenes display an antioxidant effect, we have evaluated the effect Manool, extracted from Salvia tingitana, on the extramitochondrial OxPhos and the ROI production in the retinal rod OS. Results confirm that the OxPhos machinery is ectopically expressed in the OS and that F1 Fo -ATP synthase is a target of Manool, which inhibited the OS ATP synthesis, binding the F1 moiety with high affinity, as analysed by molecular docking. Moreover, the overall slowdown of OxPhos metabolism reduced the ROI production elicited in the OS by light exposure, in vitro. In conclusion, data are consistent with the antioxidant properties of Salvia spp., suggesting its ability to lower oxidative stress production, a primary risk factor for degenerative retinal diseases. SIGNIFICANCE OF THE STUDY: Here we show that Manool, a diterpene extracted from Salvia tingitana has the potential to lower the free radical production by light-exposed rod outer segments in vitro, by specifically targeting the rod OS F1 Fo -ATP synthase belonging to the extramitochondrial OxPhos expressed on the disk membrane. The chosen experimental model allowed to show that the rod OS is a primary producer of oxidative stress linked to the pathogenesis of degenerative retinal diseases. Data are also consistent with the antioxidant and anti-inflammatory action of Salvia spp., suggesting a beneficial effect also in vivo.

A new pyrano coumarin from Clausena excavata roots displaying dual inhibition against α-glucosidase and free radical.

A new pyrano coumarin, identified as excavatin A (1) together with two known compounds nordentatin (2) and binorpocitrin (3) was isolated from the 95% EtOH extract of Clausena excavata. All structures were elucidated by using spectroscopy methods such as extensive NMR and HR-FAB-MS spectrometry. All the isolated compounds were tested on antidiabetes activity by using α-glucosidase inhibition assay and the antioxidant activity by DPPH assay. Compounds 1-3 showed antioxidant activity with IC50 values 0.286, 0.02, 0.278 mM. Among them, 2 exhibited inhibition activity against maltase (IC50 5.45 µM) and sucrase (IC50 43.57 µM). However, compounds (1) and (3) displayed inhibition on yeast α-glucosidase with IC50 values 1.92 and 5.58 mM.[Figure: see text].

Chemical Profiling of Chlorophytum comosum (Thunb.) Jaques by GC-MS/LC-ESIMS and its Antiproliferative Effects on Human Carcinoma Cell Lines.

BACKGROUND: Chlorophytum comosum, popularly known as Spider Ivy, is used as a medicinal plant in traditional Chinese medicine, however, its detailed chemical composition and biological activity are yet unexplored. OBJECTIVE: To carry out the phytochemical investigation on different parts of Chlorophytum comosum using GCMS/ LC-ESI-MS and evaluation of its antioxidant, hemolytic and antiproliferative potential on breast cancer (MCF-7), lung cancer (A549, H1299) and normal lung (L-132) cell lines. METHODS: Chemical constituents from aqueous roots and leaves extracts were identified using LC-ESI-MS/GCMS. The identified compounds were annotated based on the match of mass spectra with the literature using NIST 14 and METLIN databases. Antioxidant activity was studied using DPPH, FRAP and TPC assays. The antiproliferative effects of ethanolic roots and leaves extracts of Chlorophytum comosum were measured by MTT assay on breast cancer (MCF-7), lung cancer (A549 & H1299) and normal lung (L-132) cell lines. The toxicity studies of the extracts were carried out using Hemolysis assay. RESULTS: GC-MS analysis identified 34 metabolites in roots and 17 from leaves, while 17 compounds from roots and 7 from leaves were detected by LC-ESI-MS. Significant antiproliferative effects were observed on the A549 and MCF-7 cancer cell lines with IC50 values ranging from 56.86 µg/ml to 68.68 µg/ml while no marked response was observed against normal cell line L-132. CONCLUSION: Our study represents the first report on the detailed chemical composition and antiproliferative potential of Chlorophytum comosum against lung and breast cancer cell lines.

Crystal Guava (Psidium guajava L. "Crystal"): Evaluation of In Vitro Antioxidant Capacities and Phytochemical Content.

Free radicals can cause many diseases, such as cancer. Antioxidant is a compound that could scavenge free radicals. One of the natural antioxidants is guava. The goals of this research were to investigate the antioxidant activity of leaves and fruit of crystal guava by determining the value of the Antioxidant Activity Index (AAI) using DPPH, CUPRAC, and FRAP; evaluate the total phenolic content (TPC) and total flavonoid content (TFC); analyse the correlation between the TPC and TFC with AAI DPPH, CUPRAC, and FRAP, and analyse the correlation between the 3 methods. Extraction was performed by the reflux method using n-hexane, ethyl acetate, and ethanol. Determination of AAI DPPH, CUPRAC, FRAP, the TPC, and the TFC was performed by UV-visible spectrophotometry. The correlation of the TPC and TFC with AAI DPPH, CUPRAC, and FRAP and, also, the correlation of the 3 methods were investigated by Pearson's method. The antioxidant activity of leaves and fruit extracts of crystal guava showed AAI DPPH in the range of 0.33-56.46, CUPRAC 0.20-7.31, and FRAP 1.65-59.89. The highest TPC was given by ethanol leaf extracts (49.55 ± 1.45 g GAE/100 g), while the highest TFC was for n-hexane leaf extracts (9.68 ± 0.210 g QE/100 g). The TPC of leaves extract had a significantly positive correlation with AAI DPPH, CUPRAC, and FRAP. AAI DPPH, AAI CUPRAC, and AAI FRAP of leaves and fruit extract of crystal guava showed a significantly positive correlation. In general, leaves extract had strong antioxidant activity by the three methods. For the highest antioxidant activity, ethanol was the best solvent for extraction leaves and ethyl acetate for extraction fruit of crystal guava. The TPC in leaves extract contributed to the antioxidant activity by DPPH, CUPRAC, and FRAP methods. The Antioxidant activity of leaves and fruit extracts of crystal guava was linear by the three methods.

Tackle the free radicals damage in COVID-19.

COVID-19 is a severe pandemic which has caused a devastating amount of loss in lives around the world, and yet we still don't know how to appropriately treat this disease. We know very little about the pathogenesis of SARS-CoV-2, the virus which induces the COVID-19. However, COVID-19 does share many similar symptoms with SARS and influenza. Previous scientific discoveries learned from lab animal models and clinical practices shed light on possible pathogenic mechanisms in COVID-19. In the past decades, accumulated scientific findings confirmed the pathogenic role of free radicals damage in respiratory virus infection. Astonishingly very few medical professionals mention the crucial role of free radical damage in COVID-19. This hypothesis aims to summarize the crucial pathogenic role of free radical damage in respiratory virus induced pneumonia and suggest an antioxidative therapeutic strategy for COVID-19.

Response surface methodology for the fermentation of polysaccharides from Auricularia auricula using Trichoderma viride and their antioxidant activities.

Fermentation technology was used to improve the antioxidant activities of Auricularia auricula polysaccharide (AAP). Response surface methodology (RSM) was used to optimize the fermentation conditions. The effects of 4 independent factors: water content (X1: 40-80%), inoculation amount (X2: 2-20%), temperature (X3: 24-32 °C), and time (X4: 4-6 d) on the biological degradation efficiency were evaluated. The RSM results showed that the optimal fermentation conditions were: X1: 61.7%, X2: 12.4%, X3: 31.0 °C, X4: 5.5 d. Verification tests showed no significant differences between the practical and the predictive values for each response. Under the optimal conditions, the degradation rate was 26.89 ± 0.14%, without significant differences with the predicted value (27.03%). The degradation products were classified to different molecular weight (Mw) polysaccharide fragments using membrane separation technology. The FT-IR analysis and monosaccharide composition analysis of degraded AAP (D-AAP-VI) showed that D-AAP-VI was a furan type polysaccharide, which was different from the total AAP (pyran type). In addition, compared to total AAP, the antioxidant activities in vitro of D-AAP-VI were significantly improved (p < 0.05) and D-AAP-VI showed the strongest antioxidant activity. These results indicated that biological degradation may be a suitable way to improve the antioxidant activities of natural polysaccharides.

Minimalistic Principles for Designing Small Molecules with Multiple Reactivities against Pathological Factors in Dementia.

Multiple pathogenic elements, including reactive oxygen species, amyloidogenic proteins, and metal ions, are associated with the development of neurodegenerative disorders. We report minimalistic redox-based principles for preparing compact aromatic compounds by derivatizing the phenylene moiety with various functional groups. These molecular agents display enhanced reactivities against multiple targets such as free radicals, metal-free amyloid-ß (Aß), and metal-bound Aß that are implicated in the most common form of dementia, Alzheimer's disease (AD). Mechanistic studies reveal that the redox properties of these reagents are essential for their function. Specifically, they engage in oxidative reactions with metal-free and metal-bound Aß, leading to chemical modifications of the Aß peptides to form covalent adducts that alter the aggregation of Aß. Moreover, the administration of the most promising candidate significantly attenuates the amyloid pathology in the brains of AD transgenic mice and improves their cognitive defects. Our studies demonstrate an efficient and effective redox-based strategy for incorporating multiple functions into simple molecular reagents.

Antioxidant Content of Frozen, Convective Air-Dried, Freeze-Dried, and Swell-Dried Chokecherries (Prunus virginiana L.).

Chokecherry (Prunus virginiana L.) is rich in bioactive molecules as phenolics, which can act as antioxidants, anti-inflammatory, anticancer, among others; however, due to its high perishability, most of this fruit is wasted. Freezing and sun drying have been the most adopted techniques to avoid its postharvest deterioration. Nevertheless, both processes have presented some drawbacks as high storage costs and losses of bioactive molecules. Therefore, to preserve these molecules, this study compared the impact of convective airflow drying (CAD), freezing (FR), freeze drying (FD), and swell drying (SD). Total phenolics content (TPC), total flavonoids content (TFC), kuromanin concentration (KC), and antioxidant activity (antiradical activity (ARA) and Trolox equivalent antioxidant capacity assay (TEAC)) of chokecherries were measured. "Swell drying" is a drying process coupling convective airflow drying to the Instant Controlled Pressure Drop (DIC) expansion. A central composite rotatable design was applied to optimize the DIC variables and responses. Results showed that both freezing and swell drying effectively preserve the TPC, TFC, KC, and ARA. Moreover, SD samples also presented the highest TEAC. Contrary, in the case of CAD, it caused the highest losses of both antioxidant content and activity. Swell drying remedies the shrinkage and collapsing of dried food structure, which results in a better antioxidants extraction.

Participation of Free-Radical Processes in Structural and Metabolic Disturbances in the Lung Tissues Caused by Exposure to Coal-Rock Dust and their Adaptogenic Correction.

Adaptive correction of structural and metabolic disturbances in the lungs caused by longterm exposure to coal-rock dust were studied in experiments on rats. It was shown that the complex antioxidant preparation containing dihydroquercetin compensated disturbances in the redox balance in the lung tissue, prevented the formation of dust granulomas, and reduced the severity of degenerative changes in the bronchopulmonary system.

The apoptotic and free radical-scavenging abilities of the protein hydrolysate obtained from chicken feather meal.

This study examined the antioxidant capabilities of peptides derived from chicken feather meal (CFM) protein hydrolysates which were produced using 3 different microbial proteases (Neutrase, Alcalase, and flavourzyme) and tested at varying concentrations, namely 1, 2, and 5% by weight. The highest levels of 2,2-diphenyl-1-picrylhydrazl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activities were presented by CFM hydrolysate derived using 5 wt% Neutrase and digested for 4 h. Fractionation of this particular hydrolysate was then performed by applying 10, 5, 3, and 0.65 kDa molecular weight cutoff membranes. It was then determined that the molecular weight (MW) < 0.65 kDa fraction achieved the greatest level of free radical scavenging activity in the context of DPPH and ABTS. The MW < 0.65 kDa fraction then underwent additional fractionation using reverse-phase high-performance liquid chromatography to derive 3 main fractions designated as F1, F2, and F3. All of these fractions presented a high level of activity in DPPH radical scavenging, although no significant ABTS scavenging was observed. Quadrupole time-of-flight tandem mass spectrometry was used in determining the peptide contents of the fractions as Phe-Asp-Asp-Arg-Gly-Arg-X for F1 (FDDRGRX, 875 Da), Val-Thr-Leu-Ala-Val-Thr-Lys-His for F2 (VTLAVTKH, 868 Da), and Val-Ser-Glu-Ile-X-Ser-Ile-Pro-Ile-Ser for F3 (VSEIXSIPIS, 1,055 Da). Moreover, the F2 fraction was shown to be capable of preventing DNA damage induced by hydroxyl radicals, as indicated in tests using the plasmids pKS, pUC19, and pBR322 via the Fenton reaction. This outcome was demonstrated through in vitro antiproliferative activity in human cell lines based on SW620 colon cancer, using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. The F2 fraction at 0.5 wt.% was also shown to be capable of inducing weak early apoptosis, which could be measured by using the Fluorescein isothiocyanate Annexin V Apoptosis Detection Kit with Propidium Iodide Solution. Furthermore, an increase in caspase-3 and caspase-8 activity was observed in SW620 cells following exposure for 24 h and 48 h.

Inhibition effect of thiol-type antioxidants on protein oxidative aggregation caused by free radicals.

This study was aimed to investigate the inhibition effect of thiol-type antioxidants on protein oxidative aggregation caused by free radicals and the underlying mechanisms using six different thiol-type antioxidants (N-acetyl-L-cysteine, methionine, taurine, alpha-lipoic acid, glutathione and thioproline), Cu2+-H2O2 as a free radical generator (mainly a hydroxyl radical generator) and bovine serum albumin as the model protein. The inhibition effect of these antioxidants on protein oxidative aggregation and protective effect against oxidative damage in mouse brain tissues were investigated using SDS-PAGE, intrinsic fluorescence, simultaneous fluorescence, thioflavin T fluorescence, Congo red absorbance and inverted microscope. The results showed that all six antioxidants could inhibit protein oxidative aggregation by scavenging free radicals. In addition, alpha-lipoic acid could also bind to proteins via hydrophobic interactions and thioproline could bind to proteins via hydrogen bonds and van der Waals forces, thereby showing much stronger inhibition effect than others. Moreover, alpha-lipoic acid and thioproline could effectively prevent oxidative damage of mouse brain tissues. These results suggest that alpha-lipoic acid and thioproline can effectively inhibit free radical-induced protein aggregation and brain damage, which are worth testing for further anti-Alzheimer properties.