RESUMO
Decades of research provide evidence that certain phytochemicals in tea (Camellia sinensis) and other herbal beverages are protective against the development of sporadic types of dementia in later life. Since tea drinking is an economical and widely adopted social-cultural practice across all age groups, it is an ideal product to target in designing low-cost dietary interventions for Alzheimer's Disease (AD), the most prevalent form of dementia. In this review, we focus on the protective roles of tea-derived polyphenols and other phytochemicals on mood, the stress response, attention, and sleep, in keeping with the perspective that many early neuropathological events in AD may stem, in part, from allostatic overload. This approach aligns with the perspective that many forms of dementia, including AD, begin to take root in the brain decades prior to symptom onset, underscoring the need for early uptake of accessible and viable lifestyle interventions. The findings reviewed here suggest that consuming green and oolong tea can improve mood and reduce overall stress. However, given the caffeine content in tea and its association with stress reactivity, the effects of daily whole tea consumption on the emotional state are likely dose-dependent with an inverted-U relationship to wellbeing. Plant-based beverages that are to be consumed in high daily quantities for health purposes and which are naturally free of caffeine, such as Rooibos, may be more appropriate as a dietary supplement for managing emotional regulation over the lifetime.
Assuntos
Doença de Alzheimer , Cafeína , Humanos , Cafeína/farmacologia , Afeto , Homeostase , CháRESUMO
The brain responds to diabetic stress by inducing the inflammatory response. Under normal circumstances this process is tightly regulated. However, uncontrolled inflammatory responses lead to compromised function and eventual neurodegeneration. The microRNA (miR)-200 family, specifically miR-141, is differentially expressed in diseased states including cognitive decline, thereby triggering changes in downstream genes. We hypothesised that Metformin (MF) regulates the miR-141/protein phosphatase 2A (PP2A) axis, and associated NF-ĸB-mediated inflammasome expression in diabetic mice brain. Diabetes was induced by intraperitoneal injection of Streptozotocin (STZ), thereafter mice were treated with MF (20 mg/kg BW). Whole brain tissue was harvested for further analysis. In silico analysis showed that Sirt1 and PP2A are prediction targets of miR-141. Selected protein and gene expressions were established through western blotting and qPCR, respectively. Diabetic mice brain tissue demonstrated overexpression of miR-141 and related pro-inflammatory factors as well as decreased PP2A gene expression. MF was able to counteract this by regulating expression of miR-141, PP2A, and p-tau at Ser396 protein expressions. Further experimentation revealed MF's inhibitory action on the inflammasome system by regulating the expression of the upstream controller NLRP3, related cytokines and NF-κB signalling pathway. Collectively, we demonstrate that MF promotes neuroprotection in diabetic mice by dampening inflammatory responses through its inhibitory effects on various signalling pathways. CATEGORIES: Inflammation and Immunopharmacology, Metabolic Disorders and Endocrinology, Neuropharmacology.
Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Inflamassomos/metabolismo , Inflamação/prevenção & controle , Metformina/farmacologia , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Biologia Computacional , Diabetes Mellitus Experimental/metabolismo , Inflamação/metabolismo , Masculino , Metformina/uso terapêutico , Camundongos Endogâmicos C57BL , MicroRNAs/genética , NF-kappa B/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Proteína Fosfatase 2/genética , Proteína Fosfatase 2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/genética , Sirtuína 1/metabolismo , Estreptozocina , Proteínas tau/metabolismoRESUMO
There is a paucity of knowledge surrounding the SFC purification of human insulin. The current conventional method of insulin purification involves traditional RP-HPLC that utilises copious amounts of toxic solvents. In this study, we envisaged the development of an environmentally friendly SFC method for biosynthesized human insulin purification. Various commercially available SFC columns derived with silica, 2'ethyl pyridine, diol-HILIC, and the PFP functionalities were evaluated to determine the optimal stationary phase for purification. The PFP column gave the best results with respect to efficiencies of this important biologic that yielded average recoveries of 84%. LC-MS was used to initially detect and quantify the SFC purified standard sample of insulin (purchased) as well as the biosynthesized version. Protein sequencing was employed to verify the amino acid sequencing of the insulins; as such, the standard had a 90% probability to human insulin from the database, whereas the biosynthesized version had a 96% probability. The biological activities of both versions of the SFC purified proteins were assessed in vitro using a MTT assay. The results indicated that the biological activities of both samples were retained subsequent to SFC purification. This study successfully proposes a greener and more efficient method for the purification of insulin derivatives.
Assuntos
Cromatografia com Fluido Supercrítico/métodos , Insulina/química , Insulina/isolamento & purificação , Sobrevivência Celular , Cromatografia Líquida , Células Hep G2 , Humanos , Insulina/análise , Espectrometria de Massas , Análise de Sequência de ProteínaRESUMO
Insulin has captured researchers' attention worldwide. There is a rapid global rise in the number of diabetic patients, which increases the demand for insulin. Current methods of insulin production are expensive and time-consuming. A PCR-based strategy was employed for the cloning and verification of human insulin. The human insulin protein was then overexpressed in E. coli on a laboratory scale. Thereafter, optimisation of human insulin expression was conducted. The yield of human insulin produced was approximately 520.92 (mg/L), located in the intracellular fraction. Human insulin was detected using the MALDI-TOF-MS and LC-MS methods. The crude biosynthesised protein sequence was verified using protein sequencing, which had a 100% similarity to the human insulin sequence. The biological activity of human insulin was tested in vitro using a MTT assay, which revealed that the crude biosynthesised human insulin displayed a similar degree of efficacy to the standard human insulin. This study eliminated the use of affinity tags since an untagged pET21b expression vector was employed. Tedious protein renaturation, inclusion body recovery steps, and the expensive enzymatic cleavage of the C-peptide of insulin were eliminated, thereby making this method of biosynthesising human insulin a novel and more efficient method.
RESUMO
Diabetes is a metabolic disorder associated with mitochondrial (mt) dysfunction and oxidative stress. The molecular mechanisms involved in diabetes-associated neurological complications remain elusive. This study aims to investigate the protective effect of metformin (MF) on regulatory networks and integrated stress responses in brain tissue of Streptozotocin (STZ)-induced diabetic mice. STZ-induced diabetic mice were treated with MF (20 mg/kg BW), and whole brain tissue was harvested for further analysis. Protein carbonylation was measured as a marker of neuronal oxidative stress. Protein expression of mt chaperones, maintenance proteins, and regulators of the unfolded protein response (UPR) were measured by Western blot. Transcript levels of antioxidant enzyme GSTA4; mt biogenesis markers, ER stress regulators, and miR-132 and miR-148a were analysed using qPCR. The results showed that MF efficiently reduced protein carbonylation and oxidation. Mt function was improved by MF-treatment through upregulation of chaperone proteins (HSP60, HSP70 and LonP1). MF elicits the UPR to attenuate ER stress through a miR-132 repression mechanism. Additionally, MF was found to elevate deacetylases- Sirt1, Sirt3; and mt biogenesis marker PGC-1α through miR-148a repression. This is the first study to demonstrate the epigenetic regulation of mt maintenance by MF in diabetic C57BL/6 mouse whole brain tissue. We thus conclude that MF, beyond its anti-hyperglycaemic role, mediates neuroprotection through epigenomic and integrated stress responses in diabetic mice.
Assuntos
Encéfalo/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Metformina/farmacologia , Mitocôndrias/efeitos dos fármacos , Proteases Dependentes de ATP/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Encéfalo/patologia , Chaperonina 60/metabolismo , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/patologia , Estresse do Retículo Endoplasmático/genética , Epigênese Genética/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Masculino , Metformina/uso terapêutico , Camundongos , MicroRNAs/metabolismo , Mitocôndrias/patologia , Proteínas Mitocondriais/metabolismo , Biogênese de Organelas , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Carbonilação Proteica/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Estreptozocina/toxicidade , Resposta a Proteínas não Dobradas/efeitos dos fármacosRESUMO
The use of statins as a potential cancer drug has been investigated; however the molecular mechanisms involved in their anti-oxidant, anti-proliferative and anti-cancer effects remain elusive. In our study, we investigated the involvement of downstream mevalonate products that mediate the anti-oxidant and anti-proliferative effects of Atorvastatin (Ato), and its effect on microRNA-145 expression in HepG2 hepatocellular carcinoma cells. An amorphous soluble form of Ato was prepared and found to be cytotoxic in vitro [IC50 (1.2â¯mM); 48â¯h]. Atorvastatin induced a dose-dependent increase in cell mortality with a concomitant depletion of intracellular ATP levels (pâ¯=â¯0.005); significantly increased extracellular nitrite levels (pâ¯=â¯0.001) and decreased lipid peroxidation (pâ¯=â¯0.0097) despite a decrease in GSH. The intrinsic apoptotic pathway was activated via increased caspase -9 (pâ¯<â¯0.0001) and -3/7 (pâ¯=â¯0.0003) activities. Increased protein expression of pGSK3-(α/ß) (pâ¯=â¯0.0338), p53 (pâ¯=â¯0.0032), Mdm2 (pâ¯<â¯0.0001), with significantly diminished levels of PI3K (pâ¯=â¯0.0013), pAKT (pâ¯=â¯0.0035), and Akt (pâ¯=â¯0.0077), indicated that Ato-mediated cell death occurred via inhibition of the PI3K/Akt pathway. Additionally, the expression of PI3K (pâ¯=â¯0.0001) and c-myc (pâ¯=â¯0.0127) were also downregulated, whilst and miRNA-145 (pâ¯=â¯0.0156) was upregulated. In conclusion our data strongly indicates a plausible mechanism involved in the cytotoxic effects of Ato and is the first study to show that Ato modulates miR-145 expression in hepatocytes.â¯≤â¯.
Assuntos
Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Atorvastatina/toxicidade , MicroRNAs/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Caspases/metabolismo , Regulação para Baixo/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Glicogênio Sintase Quinase 3 beta/metabolismo , Células Hep G2 , Humanos , Malondialdeído/análise , Estresse Oxidativo/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Fator de Necrose Tumoral alfa/análise , Proteína Supressora de Tumor p53/metabolismoRESUMO
BACKGROUND: Psoriasis and psoriatic arthritis (PsA) are inflammatory associated autoimmune disorders. MicroRNA (miR)-146a plays a crucial role in regulating inflammation. A single nucleotide polymorphism in the miR-146a gene (rs2910164), aberrantly alters its gene expression and linked with the pathogenesis of several disorders, including psoriasis and PsA. In South Africa, psoriasis and PsA are extremely rare in the indigenous African population and most common in both the Indian and Caucasian population. The aim of this study was to investigate whether the miR-146a rs2910164 contributes towards psoriasis and PsA development in South African Indian and Caucasian patients. METHODS: South African Indian (n = 84) and Caucasian (n = 32) PsA patients (total n = 116) and healthy control subjects (Indian: n = 62 and Caucasian: n = 38; total n = 100) were recruited in the study. DNA was extracted from whole blood taken from all subjects, and genotyped for the miR-146a rs2910164 using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Data for laboratory parameters were obtained from pathology reports. The consulting rheumatologist collected all other clinical data. RESULTS: Unstratified data (Caucasians + Indians): A significant decrease in C-reactive protein (CRP) levels in PsA patients was observed (CRP monitored at inclusion vs. after 6 months of treatment) (18.95 ± 2.81 mg/L vs. 9.68 ± 1.32 mg/L, p = 0.0011). The miR-146a rs2910164 variant C-allele frequency in PsA patients was significantly higher vs. healthy controls (35.78% vs. 26% respectively, p = 0.0295, OR = 1.59 95% CI 1.05-2.40). Stratified data (Indians): The variant C-allele frequency in Indian PsA patients was significantly higher vs. healthy Indian controls (35.71% vs. 22.58%, p = 0.0200, OR = 1.91 95% CI 1.13-3.22). Stratified data (Caucasians): The variant C-allele frequency distribution between Caucasian PsA patients and healthy Caucasian controls was similar. CONCLUSION: The rs2910164 variant C-allele may play a role in the progression of PsA in the South African Indian population. The main limitation in this study was the small sample size in the case-control cohorts, with a low overall statistical power (post-hoc power analysis = 19%).