Evaluation of Neuroprotective Effect of Ceftriaxone on Streptozotocin Induced Alzheimer's Disease In Wistar Rats

Abstract

BACKGROUND AND OBJECTIVE Alzheimer’s disease is a progressive neurodegenerative disorder which is irreversible in nature that gradually impairs memory and thinking aptitudes and, inevitably, the capacity to complete the daily grinds. Symptoms initially show up in vast majority in their mid-60s with this disease, especially the ones with the late onset condition. The rarest condition is when the onset appear between an individual’s 30s and mid-60s, which is termed as early onset Alzheimer’s. In 2016 43.8 million and in 2017 44.9 million were the total prevalent dementia cases. Out of overall deaths in 2016, 4.4% deaths were estimated due to dementia. The number of deaths of women were more as compared to deaths of men from dementia in 2016 (1.8 vs 0.8 million). 2016 round of GBD included 195 countries and territories where 16.8 million men and 27 million women lived with dementia in 2016. In 2017 there were total 3.2 million AD cases in India. In India the total number of deaths were 0.18 million due to AD. As compared to female (1.7 million), male (1.4 million) had less prevalent cases of AD, where as the death toll of male (0.07 million) was less than female (0.10 million). By 2050, around 100 million people would be living with dementia. Ceftriaxone (CEF) which is a cephalosporin antibiotic was approved for clinical use in 1984 by FDA as a broad spectrum antibiotic. Neuronal protective reaction of CEF were noticed in animal models of neurodegenerative disorders, where CEF prohibits motor and cognitive loss; hinders dopaminergic (DAergic) decay in the striatum and substantia nigra pars compacta (SNc); mitigate cell damage in the hippocampus; recover/improve neuronal density and activity in the striatum, SNc, and hippocampus and escalates neurogenesis in the substantia nigra and hippocampal dentate gyrus (DG). Advanced glutamate reuptake through intensified glutamate transporter-1 (GLT-1) expression, boosted neurogenesis, and co-ordination of neuronal electrical action may direct the CEF intervened neuronal and behavioural protections. In patients with neuronal disorder, the outcome reinforce the utilization of CEF. Certain neurological investigations of CEF were influenced by a revolutionary discovery reported by Rothstein et al., who screened antibiotics with the β-lactam structure from 1,040 FDA approved drugs and noticed that CEF is capable to stimulate transcription of the GLT-1 gene and to raise GLT-1 protein levels in astrocytes. In many neurological diseases CEF has been examined. Research has been carried out majorly in Parkinson’s disease. Others include pain, Alexander’s disease, seizure, amyotrophic lateral sclerosis, ischemia. The aim of this study was to evaluate the effect of Ceftriaxone (CEF) on neuroprotective effect and to find out the pathogenic mechanism for that on STZ induced AD animals. MATERIALS AND METHODS In the present study, after acclimatization and training wistar rats were divided in six groups: normal control (NC), sham control (SC), disease control (DC), standard treatment (STD), ceftriaxone treated 100 mg (CEF100), ceftriaxone treated 200 mg (CEF200). Alzheimer’s disease was induced by streptozotocin (STZ)(3 mg/kg) intracerebroventricularly. Donepezil (standard) and ceftriaxone (test) treatments were given for 28 days. Normal control animals received distilled water. Disease control animals were induced by STZ. Furthermore, rats were given training on water maze and Y-maze to check the behavioural patterns and were assessed for the same. On 28th day, after the last treatment dose the animals were sacrificed, blood was collected and the brain was isolated to check the biochemical and histopathologicals changes. RESULTS The body weight changes were evaluated for 4 weeks for all the groups of the study. The body weight significantly increased in STD group (P<0.0001), CEF100 (P<0.05) and CEF200 (P<0.001) and was significantly reduced in DC group (P<0.0001) as compared to NC group. In Y-Maze, number of entries and time spent in novel arm were significantly increased in standard group (P<0.0001), CEF100 (P<0.0003), CEF200 (P<0.0001) as compared to DC group. In Morris water maze test, time to reach platform significantly declined in STD (P<0.0001), CEF100 (P<0.0001), CEF200 (P<0.0001) and time spent in target quadrant significantly increased in STD (P<0.0001), CEF100 (P<0.0001), CEF200(P<0.0001) as compared to DC group. In Acetylcholinesterase Activity significant decrease was observed in standard group (p<0.0001), CEF200 (p<0.0001) and CEF100 (p<0.0001) as compared to DC group. In total protein estimation standard treatment group (p<0.0001) and CEF100 (p<0.0001) and CEF200 (p<0.0001) showed significant decline as compared to DC group. In C-reactive protein level standard group (p<0.0001) and CEF200 (p<0.0001) showed a significant decline as compared to DC group. In reduced glutathione levels standard group (p<0.0001) and CEF200 group (p<0.0001) showed significant increase as compared to DC group. In malondialdehyde estimation, standard group (p<0.0001) and CEF200 group (p<0.0001) showed significant decline as compared to DC group. In superoxide dismutase level standard group (p<0.0001) and the CEF200 group and CEF100 group showed significant increase (p<0.0001) as compared to DC group. Histopathology was carried out with 3 stains: hematoxylin, thioflavin T and congo red. In HE stain intact pyramidal cells were found in standard group and CEF200 group as compared to DC group. In thio T stain the amyloid accumulation was not observed in standard group and CEF200 group as compared to DC group. In congo red stain the amyloid patches were not observed in standard group and CEF200 group as compared to DC group. CONCLUSION The present study demonstrates that the neurotoxin agent STZ on icv administration could develop alterations in cognitive capabilities and manifested hallmarks of Alzheimer’s disease. This cognitive alterations could be determined by neurobehavioral parameters along with biochemical parameters and histological studies. On exposure with the Standard (Donepezil) and Test drug (Ceftriaxone), the neurobehavioral and biochemical parameters showed neuroprotective effects. Therefore, from results of behavioral parameters and biochemical investigations along with histological studies it could be concluded that Ceftriaxone prevents memory impairment in rat model and can be attributable for amelioration of cognitive deficits.