Calorie restriction potentiates epigallocatechin-3-gallate-mediated Nrf2 activation in hepatocytes of aged rats

Objective: To explore the combinatorial effect of epigallocatechin-3-gallate (EGCG) and calorie restriction on activation of nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor involved in the antioxidant defense system of aged rats. Methods: Aged male Wistar rats were calorie-restricted and treated with EGCG orally for 45 days. The initial body weight of aged rats was recorded, and the final body weight was measured at the end of the experimental period. Serum lipid and lipoprotein status, oxidative stress markers such as free radicals and malondialdehyde levels, and reduced glutathione were assessed. In addition, RT-PCR and Western blotting analyses were performed. Results: Calorie restriction potentiated the effect of EGCG on enhancing antioxidant status, improving the levels of serum lipid and lipoproteins, upregulating Nrf2 and Bcl2, and downregulating Keap1, cullin3, Bax and cytochrome c in aged rats. Conclusions: Calorie restriction can promote EGCG-mediated Nrf2 activation in aged rats. This preliminary finding paves the way for a combinatory approach to replenishing the antioxidant status during aging, thereby reducing the risk for age-associated degenerative diseases

Homeopathic preparation of Allium sativum abrogates OxLDL mediated atherogenic events in macrophages: An in vitro and in silico approach

Background: Oxidized LDL (OxLDL), the key player in atherogenesis modulates endothelial dysfunction, initiates monocyte recruitment, accentuates foam cell formation, and flares up inflammatory and apoptotic events. Even though homeopathic preparation of Allium sativum has been proved to be an anti-inflammatory, anti-apoptotic and anti-atherogenic agent, its mechanism of action on abrogating OxLDL mediated foam cell formation is yet to be explored. Objective: This study was designed to bring out the role of homeopathic preparation of Allium sativum in curbing OxLDL mediated cellular inflammation in IC-21 cells exposed with OxLDL. Materials and methods: OxLDL was used to induce oxidative damage in the IC-21 macrophage cells. Assessment of inflammatory cytokines, localization of NFκB, detection of apoptosis and the in silico analysis were performed in this study. Results: The current study portrays the efficacy of homeopathy medicine as an anti-inflammatory agent, in reducing the levels of inflammatory cytokines and its mRNA expression, suppressing the activity of NFκB and preventing apoptosis in OxLDL treated IC-21 cells. Conclusion: To conclude, homeopathic preparation of Allium sativum 6C and 30C potencies are capable of controlling the transcriptional activity of NFκB and apoptosis in IC-21 cells exposed to OxLDL. These results implicate that Allium sativum homeopathic drug can be used as anti-inflammatory agent in reducing atherogenic events as it is capable of preventing OxLDL-mediated injury to macrophages

A rare genetic mutation in a stone former

A 30-year-old woman with history of passage of stones since childhood presented with oliguria and pedal edema for 10 days. She had hypertension with a creatinine of 4.1 mg/dL. Evaluation showed presence of bilateral multiple renal calculi with features of chronicity of kidney disease. Metabolic work-up for nephrolithiasis turned out to be negative and eventually renal biopsy revealed features of chronic interstitial nephritis with greenish brown refractile crystals in the tubular lumen and interstitium. The possibility of dihydroxy adenine crystalline nephropathy was considered. Spectrophotometry of RBC lysates revealed decreased activity of Adenine phosphoribosyl-transferase enzyme. Gene amplification by PCR and sequential analysis identified a missense mutation in exon 3 region of APRT gene in the patient and her family members. This case report highlights the need to contemplate the diagnosis of DHA crystalline nephropathy in young patients with nephrolithiasis and the identification of a rare genetic mutation, which is being reported for the first time in India

Homoeopathic preparation of Berberis vulgaris as an inhibitor of Calcium oxalate crystallization: An in vitro evidence

Background: Berberis vulgaris is the most widely used drug in Homoeopathy for treating urolithiasis. However, its mechanism of action in alleviating its consequences remains uncertain. Objective: To explicate the potential role of Homoeopathic preparation of B. vulgaris on in vitro Calcium oxalate (CaOx) crystallization. Materials and Methods: Spectrophotometric crystallization assay was carried out, and the slopes of the nucleation (till the maximum) and aggregation (after the peak) phases were calculated using linear regression analysis, and the percentage inhibition exerted by the modifiers was calculated. Light microscopic observation of CaOx crystals formed in the presence or absence of modifiers was carried out to support the outcome with spectrophotometric crystallization assays and to ascertain the potential role of B. vulgaris in CaOx crystallization. Results: The crystallization studies performed so far signifies B. vulgaris to be a potent drug against CaOx crystallization both at the level of nucleation and aggregation. Conclusion: Our present findings add up to the experimental evidence to support the efficacy of the homeopathic preparation of the B. vulgaris in modulating the primary events of stone formation

Targeting the Nrf2/ARE Signalling Pathway to Mitigate Isoproterenol-Induced Cardiac Hypertrophy: Plausible Role of Hesperetin in Redox Homeostasis

Cardiac hypertrophy is the underlying cause of heart failure and is characterized by excessive oxidative stress leading to collagen deposition. Therefore, understanding the signalling mechanisms involved in excessive extracellular matrix deposition is necessary to prevent cardiac remodelling and heart failure. In this study, we hypothesized that hesperetin, a flavanone that elicits the activation of Nrf2 signalling and thereby suppresses oxidative stress, mediated pathological cardiac hypertrophy progression. A cardiac hypertrophy model was established with subcutaneous injection of isoproterenol in male Wistar rats. Oxidative stress markers, antioxidant defense status, and its upstream signalling molecules were evaluated to discover the impacts of hesperetin in ameliorating cardiac hypertrophy. Our results implicate that hesperetin pretreatment resulted in the mitigation of oxidative stress by upregulating antioxidant capacity of the heart. This curative effect might be owing to the activation of the master regulator of antioxidant defense system, known as Nrf2. Further, analysis of Nrf2 revealed that hesperetin enhances its nuclear translocation as well as the expression of its downstream targets (GCLC, NQO1, and HO-1) to boost the antioxidative status of the cells. To support this notion, in vitro studies were carried out in isoproterenol-treated H9c2 cells. Immunocytochemical analysis showed augmented nuclear localization of Nrf2 implicating the action of hesperetin at the molecular level to maintain the cellular redox homeostasis. Thus, it is conceivable that hesperetin could be a potential therapeutic candidate that enhances Nrf2 signalling and thereby ameliorates pathological cardiac remodelling

Morinda citrifolia and Its Active Principle Scopoletin Mitigate Protein Aggregation and Neuronal Apoptosis through Augmenting the DJ-1/Nrf2/ARE Signaling Pathway

Given the role of oxidative stress in PD pathogenesis and off-target side effects of currently available drugs, several natural phytochemicals seem to be promising in the management of PD. Here, we tested the hypothesis that scopoletin, an active principle obtained from Morinda citrifolia (MC), efficiently quenches oxidative stress through DJ-1/Nrf2 signaling and ameliorates rotenone-induced PD. Despite reducing oxidative stress, the administration of MC extract (MCE) has lessened protein aggregation as evident from decreased levels of nitrotyrosine and α-synuclein. In vitro studies revealed that scopoletin lessened rotenone-induced apoptosis in SH-SY5Y cells through preventing oxidative injury. Particularly, scopoletin markedly upregulated DJ-1, which then promoted the nuclear translocation of Nrf2 and transactivation of antioxidant genes. Furthermore, we found that scopoletin prevents the nuclear exportation of Nrf2 by reducing the levels of Keap1 and thereby enhancing the neuronal defense system. Overall, our findings suggest that scopoletin acts through DJ-1-mediated Nrf2 signaling to protect the brain from rotenone-induced oxidative stress and PD. Thus, we postulate that scopoletin could be a potential drug to treat PD