Chronotherapeutic neuroprotective effect of verapamil against lipopolysaccharide-induced neuroinflammation in mice through modulation of calcium-dependent genes.

BACKGROUND: Neuroinflammation is a major mechanism in neurodegenerative diseases such as Alzheimer's disease (AD), which is a major healthcare problem. Notwithstanding of ample researches figured out possible molecular mechanisms underlying the pathophysiology of AD, there is no definitive therapeutics that aid in neuroprotection. Therefore, searching for new agents and potential targets is a critical demand. We aimed to investigate the neuroprotective effect of verapamil (VRP) against lipopolysaccharide (LPS)-induced neuroinflammation in mice and whether the time of VRP administration could affect its efficacy. METHODS: Forty male albino mice were used and were divided into normal control, LPS only, morning VRP, and evening VRP. Y-maze and pole climbing test were performed as behavioral tests. Hematoxylin and eosin together with Bielschowsky silver staining were done to visualize neuroinflammation and phosphorylated tau protein (pTAU); respectively. Additionally, the state of mitochondria, the levels of microglia-activation markers, inflammatory cytokines, intracellular Ca2+, pTAU, and Ca2+-dependent genes involving Ca2+/ calmodulin dependent kinase II (CAMKII) isoforms, protein kinase A (PKA), cAMP response element-binding protein (CREB), and brain-derived neurotrophic factor (BDNF), with the level of VRP in the brain tissue were measured. RESULTS: LPS successfully induced neuroinflammation and hyperphosphorylation of tau protein, which was indicated by elevated levels of microglia markers, inflammatory cytokines, and intracellular Ca2+ with compromised mitochondria and downregulated CAMKII isoforms, PKA, CREB and BDNF. Pretreatment with VRP showed significant enhancement in the architecture of the brain and in the behavioral tests as indicated by the measured parameters. Moreover, morning VRP exhibited better neuroprotective profile compared to the evening therapy. CONCLUSIONS: VRP highlighted a multilevel of neuroprotection through anti-inflammatory activity, Ca2+ blockage, and regulation of Ca2+-dependent genes. Furthermore, chronotherapy of VRP administration should be consider to achieve best therapeutic efficacy.

Protective role of vitamin D3 in a rat model of hyperthyroid-induced cardiomyopathy.

Background and aim: Several studies have reported the cardioprotective effect of vitamin D. Thus, this study aimed to investigate the possible cardioprotective effect of vitamin D3 in hyperthyroid-induced cardiomyopathy rat model. Experimental procedure: Rats were divided into 3 groups: control group; hyperthyroid group, rats were administrated l-thyroxine sodium daily for 4 weeks; and hyperthyroid + vitamin D3 treated group, rats were treated with l-thyroxine sodium for 4 weeks daily, and received the vitamin D3 for the same duration. After 4 weeks, electrocardiogram (ECG) was recorded. Then, blood samples were collected for biochemical analysis. After that, the final body weight was measured, and the rats were sacrificed. Finally, the hearts were excised, weighed and were prepared for histological examination by hematoxylin and eosin, and immunohistochemistry assessment of caspase-3 and proliferating cell nuclear antigen (PCNA). Results: Hyperthyroid rats showed significant ECG changes, increased serum levels of cardiac biomarkers, fibroblast growth factor-23 (FGF23), malondialdehyde, antioxidant enzymes, tumor necrosis factor-alpha (TNF-α) and relative heart weight compared with the control rats. Vitamin D3 coadministration with l-thyroxine resulted in significant improvement in thyroid profile, ECG parameters, significant decrease of cardiac biomarkers, FGF23, malondialdehyde, TNF-α and relative heart weight, and significant decrease of the antioxidant enzymes compared with the hyperthyroid rats. The histological study was consistent with the biochemical results. Hyperthyroid rats showed upregulation of caspase-3 and PCNA in the myocardium compared with control group. Vitamin D3 treated rats showed downregulation of caspase-3 and PCNA. Conclusion: Vitamin D3 provides cardioprotective effects in hyperthyroid rats.

Combination of magnesium supplementation with treadmill exercise improves memory deficit in aged rats by enhancing hippocampal neurogenesis and plasticity: a functional and histological study.

This study aimed to investigate the possible ameliorative effects of co-supplementation with Mg2+ and treadmill exercise on memory deficit in aged rats. Fifty male albino rats (10 young and 40 aged rats) were divided into 5 groups (10 rats/group): young, aged sedentary, aged exercised, aged Mg2+-supplemented, and aged exercised and Mg2+-supplemented. Memory was assessed using the Y-maze and novel object recognition tests. Plasma samples were collected for measurement of C-reactive protein (CRP). Subsequently, brain malondialdehyde and catalase levels were measured. Histological and immunohistochemical analyses of the hippocampi were performed. Our results showed impaired memory in aged sedentary rats, with significantly elevated plasma CRP and brain malondialdehyde levels and decreased brain catalase. The hippocampus of aged sedentary rats showed cellular degeneration, downregulation of synaptophysin (SYP) and proliferating cell nuclear antigen (PCNA), and upregulation of glial fibrillary acidic protein (GFAP) and caspase-3. Mg2+ supplementation and/or treadmill exercise significantly improved memory tests in aged rats, which could be explained by the upregulation of hippocampal SYP and PCNA expression and downregulation of GFAP and caspase-3 expression with antioxidant and anti-inflammatory mechanisms. The combined therapy had a better effect than both treatments alone, confirming the role of Mg2+ supplementation with physical exercise in enhancing age-related memory deficit. Novelty: Magnesium supplementation with treadmill exercise improves memory deficit in aged rats. The possible mechanisms are upregulation of the hippocampal synaptophysin and PCNA, downregulation of GFAP and caspase-3, the antioxidant and anti-inflammatory mechanisms.