Cell Protective Activity Research Articles

Beta cell protective effect of Curcuma longa and Piper nigrum in cytokine cocktail induced apoptosis in Min6 pancreatic beta cells

Background: Type 1 diabetes mellitus is characterized by loss of beta cell mass and insulin insufficiency. Beta cell apoptosis and destruction by cytotoxic T lymphocytes and pro-inflammatory cytokines like IL-1β, TNF-α and IFN-γ appears to represent a key event in pathogenesis of type 1 diabetes mellitus. Purpose: The aim of present study was to evaluate the prophylactic effect of aqueous extract of Curcuma longa and Piper nigrum (CL-PN) in ratio 2:1 on pancreatic beta cell apoptosis and dysfunction induced by cytokine cocktail of IL-1β (50 ng/ml), TNF-α (25 ng/ml) and IFN-γ (25 ng/ml). Methods: We used Min6 beta cell line as an in vitro model and the cells were pre-treated with herbal extract of CL-PN and then exposed to cytokine cocktail for 6 hrs to mimic beta cell destruction as in type 1 diabetes. Beta cell apoptosis and protective activity were assessed by flow cytometer based assay of annexin-V FITC and propidium iodide staining. Results: Aqueous extract of the combination of CL-PN at 1 μg/ml concentration can effectively inhibit apoptosis in Min6 beta cell line by 1.6 fold as compared to cytokine cocktail. It also demonstrated 2.6 fold lesser production of nitric oxide, 1.4 fold reduced ROS and increasing mitochondrial membrane potential which are major markers of beta cell damage. The caspase 3 mRNA expression is also reduced by 4.7 fold despite cytokine challenge. Conclusion: Therefore, the present study paves the way for establishing herbal oral treatment for protection of beta cells in type 1 diabetes mellitus.

Aspalathus linearis infusion affects hole-board test behaviour and amino acid concentration in the brain

Rooibos tea, brewed using Aspalathus linearis leaves, is a popular South African herbal infusion, but its everyday intake is not fully described in terms of the neuropsychopharmacological outcomes. The cell-protective activity of A. linearis is connected with the ability of reducing glycaemia, inflammation as well as oxidative stress. It was already shown that “fermented” rooibos herbal tea (FRHT), which is rich in phenolic compounds, improves the cognitive performance of rats in the water maze and impacts dopaminergic striatal transmission. The present research was taken to extend the knowledge about the feasible behavioural and neurochemical implications of sustained oral FRHT consumption. We hypothesized that it might affect brain amino acid content and thus induce behaviour and neuroprotection.FRHTs of different leaf to water ratios (1:100, 2:100 and 4:100), analysed by chromatographic methods as regards their flavonoid characteristics, were given to rats as only liquid for 3 months. Their behaviour was evaluated in the hole-board test (HBT). Brain amino acids concentration was analysed in the striatum, hippocampus and prefrontal cortex by HPLC-ECD. The rats drinking rooibos tea presented increased motor activity defined as time spent on moving in the HBT. Their exploration measured by head-dipping and rearing was enhanced. Longer time of the testing-box central zone occupation indicated to reduction in anxiety-related behaviour. Excitatory amino acids (aspartate and glutamate) content was decreased in the striatum of animals drinking the infusions whereas taurine level was increased both in the striatum and hippocampus. In conclusion we suggest that long-term FRHT intake affects exploration and anxiety-related behaviour of the rats as well as exerts biochemical outcomes in the brain that support the neuroprotective impact of rooibos tea.

Surface display of p75, a\xa0Lactobacillus rhamnosus GG derived protein, on Bacillus subtilis spores and its antibacterial activity against Listeria monocytogenes

Lactobacillus rhamnosus p75 protein with peptidoglycan hydrolase (PGH) activity is one of the key molecules exhibiting anti-apoptotic and cell-protective activity for human intestinal epithelial cells. In this study, with the goal of developing new probiotics, the p75 protein was displayed on the surface of Bacillus subtilis spores using spore coat protein CotG as an anchoring motif. The PGH activity, stability, and the antibacterial activity of the spore-displayed p75 (CotG-p75) protein were also investigated. The PGH activity of the CotG-p75 against peptidoglycan extracted from B. subtilis was confirmed by the ninhydrin test. Under various harsh conditions, compared to the control groups, the PGH activities of CotG-p75 were very stable in the range of pH 3–7 and maintained at 70% at 50 °C. In addition, the antibacterial activity of CotG-p75 against Listeria monocytogenes was evaluated by a time-kill assay. After 6 h incubation in phosphate-buffered saline, CotG-p75 reduced the number of viable cells of L. monocytogenes by up to 2.0 log. Scanning electron microscopy analysis showed that the cell wall of L. monocytogenes was partially damaged by the treatment with CotG-p75. Our preliminary results show that CotG-p75 could be a good candidate for further research to develop new genetically engineered probiotics.

Poly(gallic acid)-coated polycaprolactone inhibits oxidative stress in epithelial cells.

Enzymatic mediated poly (gallic acid) (PGAL), a stable multiradical polyanion with helicoidal secondary structure and high antioxidant capacity, was successfully grafted to poly(ε-caprolactone) (PCL) using UV-photo induction. PCL films were prepared with several levels of roughness and subsequently grafted with PGAL (PCL-g-PGAL). The results on the full characterization of the produced materials by mechanical tests, surface morphology, and topography, thermal and crystallographic analyses, as well as wettability and cell protection activity against oxidative stress, were adequate for tissue regeneration. The in vitro biocompatibility was then assessed with epithelial-like cells showing excellent adhesion and proliferation onto the PCL-g-PGAL films, most importantly, PCL-g-PGAL displayed a good ability to protect cell cultures on their surface against reactive oxygen species. These biomaterials can consequently be considered as novel biocompatible and antioxidant films with high-responsiveness for biomedical or tissue engineering applications.

Two distinct ubiquitin-binding motifs in A20 mediate its anti-inflammatory and cell-protective activities.

Protein ubiquitination regulates protein stability and modulates the composition of signaling complexes. A20 is a negative regulator of inflammatory signaling, but the molecular mechanisms involved are ill understood. Here, we generated Tnfaip3 gene-targeted A20 mutant mice bearing inactivating mutations in the zinc finger 7 (ZnF7) and ZnF4 ubiquitin-binding domains, revealing that binding to polyubiquitin is essential for A20 to suppress inflammatory disease. We demonstrate that a functional ZnF7 domain was required for recruiting A20 to the tumor necrosis factor receptor 1 (TNFR1) signaling complex and to suppress inflammatory signaling and cell death. The combined inactivation of ZnF4 and ZnF7 phenocopied the postnatal lethality and severe multiorgan inflammation of A20-deficient mice. Conditional tissue-specific expression of mutant A20 further revealed the key role of ubiquitin-binding in myeloid and intestinal epithelial cells. Collectively, these results demonstrate that the anti-inflammatory and cytoprotective functions of A20 are largely dependent on its ubiquitin-binding properties.

Correction to: Protein-rich extract of Musca domestica larvae alleviated metabolic disorder in STZ-induced type 2 diabetic rat model via hepatoprotective and pancreatic β-cell protective activities

In the December 2018 issue of the Journal of Biosciences, in the article titled ‘‘Protein-rich extract of Musca domestica larvae alleviated metabolic disorder in STZ-induced type 2 diabetic rat model via hepatoprotective and pancreatic β -cell protective activities’’ by Hanfang Mei et al. (DOI: 10.1007/s12038-018-9804-z; Vol. 43, No. 5, pp. 969–983), the affiliations of Hanfang Mei have been incompletely mentioned as:

Binding to carboxypeptidase M mediates protective effects of fibrinopeptide Bβ15-42

During fibrinolysis a 28-amino-acid peptide is generated besides other degradation products of fibrin. This peptide, called Bβ15-42, which is cleaved by plasmin from the end of the fibrin Bβ-chain, is protective in myocardial and renal ischemia/reperfusion injury and improves the outcome in experimental sepsis. Bβ15-42 has been shown to mediate different beneficial effects in endothelial cells through binding to vascular endothelial-cadherin. Here, we provide in vitro and in vivo evidence that Bβ15-42 has additional cell protective activity in tubular cells, which is caused by a distinct mechanism. As vascular endothelial-cadherin is not expressed by tubular cells we used ligand-receptor capture technology LRC-TriCEPS to search for tubular cell surface receptors and identified carboxypeptidase M (CBPM) as a novel binding partner of Bβ15-42. Silencing CBPM with siRNA reduced the protective potential of Bβ15-42 against tubular cell stress. Bβ15-42 inhibited the enzymatic activity of CBPM and modified the impact of CBPM on bradykinin signaling. We conclude that beneficial properties of Bβ15-42 are not restricted to endothelial cells but are also active in epithelial cells where cytoprotection depends on CBPM binding.

Dissection of mechanisms of Chinese medicinal formula Si-Miao-Yong-an decoction protects against cardiac hypertrophy and fibrosis in isoprenaline-induced heart failure.

Si-Miao-Yong-An decoction (SMYAD) is a traditional Chinese herbal formulation. SMYAD first appeared in the Eastern Han Dynasty according to the "Shen Yi Mi Zhuan". Then the formula was recorded in the "Yan Fang Xin Bian" edited by medical scientist Bao Xiangao in the Qing Dynasty. This well-known prescription has been traditionally used for gangrene and vascular vasculitis. It is mainly used for cardiovascular and endocrine diseases in current clinical applications and research. In this study, the potential mechanisms of SMYAD against cardiac fibrosis and hypertrophy in the β-adrenoceptor agonist isoprenaline induced heart failure model were investigated. The heart failure animal model was established via injected isoprenaline in rats. Echocardiography was used to detect the structure and function of the heart. HE staining and Masson's trichrome staining was performed to assess myocardial tissue morphology. The serum biochemical indexes were detected by dedicated biochemical kit. BNP was tested by ELISA kit. The levels of mRNA were detected by RT-qPCR. Cardiomyocyte morphology was assessed by immunofluorescence. Phosphorylated and total p38, Akt were analyzed by Western blot. The production of reactive oxygen species (ROS) was tested by CM-H2DCFDA probe. Formula identification of chemical constituents of SMYAD in plasma was disclosed through ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS). SMYAD was able to improve the heart function in ISO induced heart failure rat model via protecting rat from developing cardiac hypertrophy and fibrosis. SMYAD also decreased plasma expression of these biochemical indexes. It was found that SMYAD could regulate cardiac hypertrophy and fibrosis makers' mRNA levels in vitro and vivo. In addition, SMYAD inhibited the phosphorylation of p38 and Akt, which are key mediators in the pathological process of ISO-induced cardiac hypertrophy and myocardial fibrosis. It also showed that the components of SMYAD in rat plasma exerted myocardial cell protective activity. In summary, SMYAD may comprise more than one active ingredient to the pursuit of combination therapies instead of specifically target a single disease-causing molecule. These experimental results suggest that SMYAD may be a potential drug candidate in diseases of cardiac hypertrophy and myocardial fibrosis caused by β-adrenoceptor abnormalities.

Long-term administration of Aspalathus linearis infusion affects spatial memory of adult Sprague-Dawley male rats as well as increases their striatal dopamine content

Ethnopharmacological relevanceEveryday use of the herbal tea rooibos, produced from Aspalathus linearis (Brum.f) Dahlg. (Fabaceae) is customary in South Africa, a continuation of its historical use by indigenous people. Although evidence of its traditional indications is anecdotal, rooibos tea is regarded as a general health tea. Aims of the studyAvailable contemporary research indicates to broad cell protective activity of rooibos focusing on its antioxidative, anti-inflammatory, anti-hyperglycaemic and antithrombotic features affecting metabolic syndrome, cardiovascular risk and neuroprotection. Nevertheless little is known about its impact on brain functions. The present experiment aimed to evaluate the possible behavioural and neurochemical effects of long-term oral administration of “fermented”” rooibos herbal tea (FRHT) infusions to adult male Sprague-Dawley rats. Materials and methodsInfusions, prepared using 1, 2 and 4 g of “fermented”” (oxidised) A. linearis leaves for 100 ml of hot water, were characterised in terms of flavonoid content by ultra-high and high performance liquid chromatography (UHPLC-qTOF-MS, HPLC-DAD) and administered to rats as sole drinking fluid for 12 weeks. Spatial memory behaviour was assessed in a modified version of the Morris water maze. Dopamine, noradrenaline, serotonin and their metabolite levels (DOPAC, 3-MT, HVA, MHPG, 5-HIAA) were quantified in prefrontal cortex, hippocampus and striatum by HPLC-ECD. Body weight and blood glucose level were additionally estimated. ResultsAll FRHT-treated rats showed improvement of long-term spatial memory defined as increased number of crossings over the previous platform position in SE quadrant of the water maze. It was not accompanied by excessive motor activity. Striatal dopamine and its metabolite 3-MT (3-methoxytyramine) levels were increased in treated rats. There were no differences in body weight gain between control and treated animals but blood glucose level was significantly lower in the latter ones. ConclusionThe improvement of long-term memory in FRHT-treated rats and stimulating impact of FRHT on their dopaminergic striatal transmission support the wellness enhancing effect of rooibos tea, contributing to a better understanding of the neurological background of traditional habitual consumption of this herbal tea.

Simultaneous quantitative analysis of 11 flavonoid derivatives with a single marker in persimmon leaf extraction and evaluation of their myocardium protection activity.

An improved, reliable and comprehensive method for assessing the quality of the ethyl acetate extract from persimmon leaves (EAPL) and its commercial preparation, Naoxinqing (Brain and Heart Clear capsules), has been developed and validated. Based on HPLC-DAD-ESI-Q-TOF-MS analysis, myricetin-3-O-β-D-galactoside (1), myricetin-3-O-glucoside (2), quercetin-3-O-β-D-galactoside (3), quercetin-3-O-β-D-glucoside (4), quercetin-3-O-(2″-O-galloyl-β-D-galactoside) (5), quercetin-3-O-(2″-O-galloyl-β-D-glucoside) (6), kaempferol-3-O-β-D-galactoside (7), kaempferol-3-O-β-D-glucoside (8), kaempferol-3-O-(2″-O-galloyl-β-D-galactoside) (9), kaempferol-3-O-(2″-O-galloyl-β-D-glucoside) (10), quercetin (11) and kaempferol (12) were identified from 15 batch samples. A HPLC fingerprint analytical method was established. All compounds, with the exception of compound 2, were simultaneously quantified by the single standard to determine multi-components (SSDMC) method, using kaempferol-3-O-β-D-glucoside as the internal standard. The rate of analysis was found to be faster with the SSDMC method than with current acid hydrolysis method (Pharmacopoeia of the People's Republic of China 2015 edition) and the results were more intuitive and reliable. Three-dimensional principal component analysis revealed that there were similar characteristics in persimmon leaf from same district. Analysis of the myocardial cell protection activity of 11 monomeric compounds showed that compounds 12, 11 and 10 were the main active ingredients that produce pharmacologic functions in EAPL. Among these compounds, the bioactive constituent of myricetin-3-O-β-D-galactoside was determined for the first time in Diospyros khaki. Thus, we have established an effective assessment method that can be applied to the comprehensive quality evaluation of EAPL extract and Naoxinqing capsule.

RA1M-035, a Physicochemically Improved Human Recombinant α1-Microglobulin, Has Therapeutic Effects in Rhabdomyolysis-Induced Acute Kidney Injury.

Aims: Human α1-microglobulin (A1M) is an endogenous reductase and radical- and heme-binding protein with physiological antioxidant protective functions. Recombinant human A1M (rA1M) has been shown to have therapeutic properties in animal models of preeclampsia, a pregnancy disease associated with oxidative stress. Recombinant A1M, however, lacks glycosylation, and shows lower solubility and stability than A1M purified from human plasma. The aims of this work were to (i) use site-directed mutagenesis to improve the physicochemical properties of rA1M, (ii) demonstrate that the physicochemically improved rA1M displays full in vitro cell protective effects as recombinant wild-type A1M (rA1M-wt), and (iii) show its therapeutic potential in vivo against acute kidney injury (AKI), another disease associated with oxidative stress.Results: A novel recombinant A1M-variant (rA1M-035) with three amino acid substitutions was constructed, successfully expressed, and purified. rA1M-035 had improved solubility and stability compared with rA1M-wt, and showed intact in vitro heme-binding, reductase, antioxidation, and cell protective activities. Both rA1M-035 and rA1M-wt showed, for the first time, potential in vivo protective effects on kidneys using a mouse rhabdomyolysis glycerol injection model of AKI.Innovation: A novel recombinant A1M-variant, rA1M-035, was engineered. This protein showed improved solubility and stability compared with rA1M-wt, full in vitro functional activity, and potential protection against AKI in an in vivo rhabdomyolysis mouse model.Conclusion: The new rA1M-035 is a better drug candidate than rA1M-wt for treatment of AKI and preeclampsia in human patients.

Leptospermum flavescens Sm. protect pancreatic β cell function from streptozotocin involving apoptosis and autophagy signaling pathway in in vitro and in vivo case study

Ethnopharmacological importanceLeptospermum flavescens has been used traditionally in Malaysia to treat various ailments such as constipation, hypertension, diabetes and cancer. Aim of studyTo investigate the potential protective effects of L. flavescens in pancreatic β cells through inhibition of apoptosis and autophagy cell death mechanisms in in vitro and in vivo models. Materials and methodsL. flavescens leaves were extracted using solvent in increasing polarities: hexane, ethyl acetate, methanol and water. All extracts were tested for INS-1 β cells viability stimulated by streptozotocin (STZ). The extract which promotes the highest cell protective activity was further evaluated for insulin secretion, apoptosis and autophagy signaling pathways. Then, the acute toxicity of extract was carried out in SD rats according to OECD 423 guideline. The active extract was tested in diabetic rats where the pancreatic β islets were evaluated for insulin, apoptosis and autophagy protein. ResultsThe methanolic extract of L. flavescens (MELF) was found to increase INS-1 β cells viability and insulin secretion against STZ. In addition, MELF has been shown to inhibit INS-1 β cells apoptosis and autophagy activity. Notably, there was no toxicity observed in SD rats when administered with MELF. Furthermore, MELF exhibited anti-hyperglycemic activity in diabetic rats where apoptosis and autophagy protein expression was found to be suppressed in pancreatic β islets. ConclusionMELF was found to protect pancreatic β cells function from STZ-induced apoptosis and autophagy in in vitro and in vivo.

Trypsin and ascorbic acid have a synergistic effect on the quality of apple processing by protecting apple cells from oxidative damage

Flesh browning is fundamentally important to the quality of minimally processed fruit. The development of browning is associated with reactive oxygen species accumulation in the fruit tissue. The novel superoxide scavenging activity of trypsin has been shown in our previous work. In the current study, the effects of mixtures of trypsin and ascorbic acid (AsA) on the processing quality of apple homogenate were evaluated. In the presence of trypsin and AsA, the browning of apple homogenate was significantly impeded, and changes in the titration of acid and in the concentration of soluble solids were also delayed. The mixture of trypsin and AsA could significantly reduce cell membrane damage and the activity of peroxidase. Above all, trypsin could protect the cells of the fruit due to its superoxide scavenging activity. The utilization of trypsin will provide a new strategy for controlling quality in the processing of apples or other fruits and vegetables. Practical applications Pure, natural fruit juice with fruit flavor, especially pure, cloudy juice with a variety of natural nutrients, is increasingly widely accepted. Eliminate or inhibit the free radical accumulation is one of the most efficient methods to control the browning of apple tissue. Trypsin had a significant effect on the superoxide anion scavenging activity and showed a good protective effect on the cells. This article investigates the antioxidant and cell protective activity of trypsin–ascorbic acid (AsA) mixture during the processing of apple homogenate. The application of trypsin provides a new strategy for the quality control of fruit juice. Food additives with trypsin have a broad market and development potential.

OP-33 - A1M-035, an improved human recombinant alpha-1-microglobulin, has therapeutic effects in rhabdomyolysis-induced acute kidney injury

Human A1M (alpha-1-microglobulin) is a reductase and radical- and heme-binding protein with physiological antioxidant protective functions. Recombinant human A1M has been shown to have therapeutic properties in vitro and in vivo, but lacks glycosylation, and shows lower solubility and stability than A1M purified from human plasma. The aims of this work were to 1) use site-directed mutagenesis to improve the physicochemical properties of A1M, 2) demonstrate that the physicochemically improved A1M displays full in vitro cell protective effects as recombinant wild-type A1M (A1M-wt), and 3) show its therapeutic potential in vivo against acute kidney injury (AKI), a disease associated with oxidative stress, using a mouse rhabdomyolysis glycerol-injection model. A novel recombinant A1M-variant (A1M-035) with three amino acid substitutions was constructed, successfully expressed and purified. A1M-035 had improved solubility and stability compared to A1M-wt, and had intact in vitro heme-binding, reductase, antioxidation, and cell protective activities. Both A1M-035 and A1M-wt showed, for the first time, potential in vivo protective effects on acute kidney injury (AKI). It is concluded that the new A1M-035 is a better drug candidate than A1M-wt for treatment of AKI in human patients.

Antioxidant nitroxides protect hepatic cells from oxidative stress-induced cell death.

Oxidative stress causes cell death and induces many kinds of disease, including liver disease. Nitroxides are known to react catalytically with free radicals. In this study, the cell protective activities of nitroxides were compared with those of other antioxidants. Nitroxides showed much greater inhibition of hydrogen peroxide-induced cell death than other antioxidants in a hepatic cell line and in primary hepatocytes. The intracellular oxidative stress level at 24 h after hydrogen peroxide stimulation was significantly decreased by nitroxides, but not by other antioxidants. To clarify the mechanism of cell protection by nitroxides, we investigated whether nitroxides inhibited DNA damage and mitogen-activated protein kinase pathway activation. We found that nitroxides reduced caspase-3 activation and may have ultimately inhibited cell death. In conclusion, nitroxides are very useful for attenuating cell damage due to oxidative stress. Nitroxides are thus a potential therapeutic agent for oxidative stress-related diseases.

Ellagic acid protects against neuron damage in ischemic stroke through regulating the ratio of Bcl-2/Bax expression.

An oxygen-glucose deprivation and reoxygenation model in primary cultured rat cortical neurons was developed for this study to investigate the effects of ellagic acid (EA), a low-molecular-weight polyphenol, on neuron cells and their function, and to evaluate whether EA can be safely utilized by humans as a functional food or therapeutic agent. Administration of EA significantly decreased the volume of cerebrum infarction and the neurological deficit scores of the rats; EA treatment also increased the number of Bcl-2-positive cells and the ratio of Bcl-2-positive to Bax-positive neurons in the semidarkness zone near the brain ischemic focus in the photothrombotic cerebral ischemia model. Treatment of EA resulted in increased neuron viability, cell nuclear integrity, and the ratio of Bcl-2/Bax expression in the primary cultured neuron model; EA treatment also lead to a decrease in the number of apoptotic cells. Our results therefore suggest a specific mechanism for the beneficial effects of EA, providing new insights into how it provides neuroprotection. To the best of our knowledge, these results represent new insights on the mechanisms of the brain cell protective activity of EA. Thus, EA may be used in functional foods or medicines to help treat nerve dysfunction, neurodegenerative disease, and aging.

Effects of oxyresveratrol and its derivatives on cultured P19-derived neurons

Purpose: To modify the structure of oxyresveratrol and evaluate the obtained derivatives for effects on neuronal cells. Methods: Electron-withdrawing groups were selectively introduced to the aromatic ring of the core stilbene structure. Oxyresveratrol and derivatives were then evaluated for their ability to enhance the survival of P19 derived neuronal cells by XTT method, in comparison with the widely known antioxidants, Trolox and ascorbic acid. Phase-contrast microscopic images of the neurons under various conditions were also taken and analyzed. Results: Oxyresveratrol, at a very low concentration (1 ng/mL), enhanced the survival of neurons in both normal and serum-deprivation conditions. Higher activity was observed for the 5-formylated and t 5-carboxylated products. The potencies of these polyoxygenated stilbenes were far greater than those of Trolox and ascorbic acid. These observations were supported by results from the examination of the phase-contrast micrographs of the neuronal cells. Conclusion: Oxyresveratrol and some derivatives prepared in this study demonstrate significant cell protective activity and may be of therapeutic value, but further investigations in animals are required to verify their neuroprotective potentials. Keywords: Oxyresveratrol, Artocarpus lacucha , Artocarpus lakoocha , Polyoxygenated stilbenes, Cell protection, Neuron

Ellagic acid improves endogenous neural stem cells proliferation and neurorestoration through Wnt/β-catenin signaling in vivo and in vitro.

The aim of this study is to research the effects of the polyphenol ellagic acid (EA) on brain cells and to explore its mechanism of action, and to evaluate whether EA can be safely utilized by humans as a functional food or therapeutic agent. A photothrombosis-induced model of brain injury in rats was created, and EA was administered intragastrically to rats on 7 consecutive days post-venous ischemia. An oxygen-glucose deprivation and re-perfusion model was established in neural stem cells in order to research the effects on proliferation after 2 days of EA treatment in vitro. The administration of EA improved the rats' nerve-related abilities, remedied infarct volumes and morphological changes in the brain, and enhanced the content of nestin protein in the brain semidarkness zone. The proliferation of NSCs and the expression of β-catenin and Cyclin D1 genes were also increased in primary cultured NSCs. EA administration can improve brain injury outcomes and increase the proliferation of NSCs through the Wnt/β-catenin signaling pathway. The presented results represent new insights on the mechanisms of the brain cell protective activity of EA. Thus, EA may be used in functional foods or medicines to help treat nerve dysfunction, neurodegenerative disease and aging.

Microbial transformation of Pseudoprotodioscin by Chaetomium olivaceum

Pseudoprotodioscin (1) is a steroidal saponin from plants with diverse biological activities. In order to obtain new and useful derivatives for further study, Chaetomium olivaceum was used to transform pseudoprotodioscin. Five new (4, 5, 6, 8, and 9) and three known (2, 3 and 7) steroidal saponins were obtained in this study. These reactions were hard to be achieved through chemical modification. The structures of these metabolites were elucidated on the basis of spectroscopic data. The possible biotransformation pathway of 1 by Chaetomium olivaceum was also proposed. Protective effects against H2O2-induced H9c2 cardiomyocyte injury of compounds 1–9 were evaluated. Compounds 1, 4 and 6 showed some cell-protection activity.

Simplified Bryostatin Analogues Protect Cells from Chikungunya Virus-Induced Cell Death.

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus showing a recent resurgence and rapid spread worldwide. While vaccines are under development, there are currently no therapies to treat this disease, except for over-the-counter (OTC) analgesics, which alleviate the devastating arthritic and arthralgic symptoms. To identify novel inhibitors of the virus, analogues of the natural product bryostatin 1, a clinical lead for the treatment of cancer, Alzheimer’s disease, and HIV eradication, were investigated for in vitro antiviral activity and were found to be among the most potent inhibitors of CHIKV replication reported to date. Bryostatin-based therapeutic efforts and even recent anti-CHIKV strategies have centered on modulation of protein kinase C (PKC). Intriguingly, while the C ring of bryostatin primarily drives interactions with PKC, A- and B-ring functionality in these analogues has a significant effect on the observed cell-protective activity. Significantly, bryostatin 1 itself, a potent pan-PKC modulator, is inactive in these assays. These new findings indicate that the observed anti-CHIKV activity is not solely mediated by PKC modulation, suggesting possible as yet unidentified targets for CHIKV therapeutic intervention. The high potency and low toxicity of these bryologs make them promising new leads for the development of a CHIKV treatment.