Therapeutic enhancing potential of piracetam with diethylstilbestrol in prevention of grand-mal seizures in rats: inhibition of PI3K/Akt/mTOR signaling pathway and IL-1ß, IL-6, TNF-α cytokines levels.

OBJECTIVE: Epilepsy, a neurodegenerative disorder, continues to throw challenges in the therapeutic management. The current study sought to ascertain if the therapeutic interactions between piracetam and diethylstilbestrol may prevent grand-mal seizures in rats. MATERIALS AND METHODS: Piracetam (PIR; 10 and 20 mg/kg) and diethylstilbestrol (DES; 10 and 20 mg/kg) alone as a low-dose combination were administered to rats for 14 days. The electroshock (MES; 180 mA, 220 V for 0.20 s) was delivered via auricular electrodes on the last day of treatment and rats were monitored for convulsive behavior. To elucidate the mechanism, hippocampal mechanistic target of rapamycin (mTOR) and interleukin (IL)-1ß, IL-6 and tumor necrotic factor-alpha (TNF-α) levels were quantified. Hippocampal histopathology was conducted to study the neuroprotective effect of drug/s. In vitro studies and in silico studies were conducted in parallel. RESULTS: To our surprise, the low dose of the combination regimen of PIR (10 mg/kg) and DES (10 mg/kg) unfolded synergistic anti-seizure potential, with brimming neuroprotective properties. The mechanism could be related to a significant reduction in the levels of hippocampal mTOR and proinflammatory cytokines. The docking scores revealed higher affinities for phosphatidylinositol 3-kinase (PI3K) in co-bound complex, and when docking DES first, while better affinities for protein kinase B (Akt) were revealed when docking PIR first (both drugs bind cooperatively as well). This indicated that the entire PI3K/Akt/mTOR signaling pathway is intercepted by the said combination. In addition, the % of cell viability of HEK-293 cells [pre-exposed to pentylenetetrazol (PTZ)] was increased by 327.29% compared to PTZ-treated cells (toxic control; 85.16%). CONCLUSIONS: We are the first to report the promising efficacy of the combination (PIR 10 mg/kg + DES 10 mg/kg) to restrain seizures and epileptogenic changes induced by electroshock by a novel mechanism involving inhibiting the PI3K/Akt/mTOR signaling.

Lack of association between B 12 and Body Mass Index among Saudi multiple sclerosis patients.

Background: Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system. It is an autoimmune disease of multifactorial etiology, linked to a variety of genetic and well-defined environmental factors. It typically affects more women than men and more frequently affects adults aged 20-45 years. Besides, vitamin B12 deficiency and obesity are associated with exacerbating central nervous system inflammation and a higher clinical disability. Objective: The study aims to determine the association of the vitamin B12 serum concentration with the Body Mass Index BMI, thyroid-stimulating hormone serum levels and MS clinical features in Saudi MS patients. Methods and results: This is a retrospective cohort study, and data were collected from the MS database at the King Fahad Medical City Multiple Sclerosis Clinic, from December 2015 to December 2019. Data were entered and analyzed using the Statistical Package for Social Sciences (SPSS ver. 20, Chicago, IL, USA). Cobalamin, also known as vitamin B12, has a reference concentration that ranges from 138 to 652 Pmol/L in adults. The patient's BMI was calculated by dividing the weight (in kilograms) by the square of the height (in square meters), expressed in kg/m2.Data for 169 MS subjects were collected. A total 83 of them, with a mean age of 36.2 ± 9.57 years, had vitamin B12 results. Of all patients, 16.6% had vitamin B12 deficiency (<138 pmol/L) and 9.52% of them were overweight, BMI kg/m2 = (25-29.9). The mean vitamin B12 level in all MS subjects was 240 ± 117 pmol/L. Moreover, 58.33% of the MS patients had high BMIs (BMI >25). However, no significant correlation was found between vitamin B12 deficiency neither with the BMI nor TSH concentration in MS cases (r = 0.03, p = 0.64), (r = 0.00, P = 0.9) respectively. Conclusion: These findings revealed no association between serum vitamin B12 concentration and TSH, BMI in MS clinical parameters, however, further studies are required to validate these results.