T29347
انگلیسی
Investigating of histological and stereological changes along with the oxidative stress parameters and alternating of NMDA and AMPA receptor genes expression in the cerebellum of male rats during Alzheimer's disease induction and treatment with Niosome Loaded Curcumin
Dissertation
Mustafa Saleh Abed ALshiblawi
Natural sciences
1402
172p.
cd
M.S.
biology
1402/06/27
Alzheimer's disease is a disease that destroys the nervous system and is the main cause of dementia that affects memory and learning ability. This disease is the fourth most expensive disease in all high-income countries and there is still no effective treatment for this disease. There are several hypotheses for Alzheimer's pathophysiology, especially the amyloid hypothesis and the tau hypothesis. Also, the hypotheses of inflammation, oxidative stress, accumulation of heavy metals such as aluminum and high levels of copper, iron and zinc ions in the brain and the cholinergic hypothesis are also causes of this disease. Long-term exposure to aluminum chloride increased amyloid beta polymerization. Also, oxidative stress plays a key role in Alzheimer's disease by disrupting the balance of oxidative parameters and activating inflammatory features. It highlights glutamate neurotransmission through (NMDA) and (AMPA) receptors as a critical mediator of synaptic plasticity. Despite this positive role, high concentrations of extracellular glutamate lead to excitotoxicity and neurodegeneration. NMDA receptors containing GluN2A subunits are located at synaptic sites involved in protective pathways. In contrast, GluN2B-containing receptors are mainly located at extrasynaptic sites and increase neuronal vulnerability. AMPA receptors are continuously inserted into the membrane and recycled. An increased rate of cellular endocytosis is implicated as part of the pathophysiology of Alzheimer's disease by inducing long-term depression (LTD) and synaptic breakdown. Curcumin belongs to a class of compounds called curcuminoids and has neuroprotective properties, in such a way that it improves Alzheimer's disease by reducing inflammation and modulating oxidative stress. The brain is absorbed. Therefore, nanoparticles (nano drug carriers) are used in order to increase the bioavailability and better performance of antioxidants. Niosomes are a group of nanoparticles with a structure of a double surfactant layer, which, due to having hydrophobic and hydrophilic ends, perform well in carrying antioxidants. and drugs can have, therefore this nanoparticle can be used as a carrier of curcumin antioxidant in the treatment of Alzheimer's disease. Therefore, the purpose of this study is to investigate the histological and stereological changes along with the oxidative stress parameters and changes in the expression of NMDA and AMPA genes in Cerebellum of male rats during induction of Alzheimer's disease and treatment with curcumin nanoparticles.Materials and methods:56 male Wistar rats were randomly assigned into 7 groups (n=8). Saline (1 ml/kg), Ethanol 9% (1 ml/kg), Alzheimer’s disease (AD) induction group with Aluminum chloride (4.2 mg/kg; i.p for 28 days), Curcumin (20 mg/kg; i.p for 14 days), Curcumin (20 mg/kg; i.p for 7 days), Alzheimer’s disease + Curcumin (20 mg/kg; i.p for 14 days), Alzheimer’s disease + Curcumin (20 mg/kg; i.p for 7 days). Rota-Rod and grip strength tests were used for studing neuromuscular parameters of rats. After receiving treatments, the animals were decapitated and their cerebellum were dissected and divided into 2 right and left hemispheres. Left hemisphere divided into 2 parts, one part for investigating the expression of AMPA and NMDA genes expression using RT- PCR and one part for assessing oxidative stress parameters. These parts instantly stored at – 80 until use. Right hemisphere of cerebellum, stored in formalin 15% for analyzing histological and stereological parameters. One-way analysis of Variance (ANOVA) and post-hoc Tukey test were used for data analyzing.Results: The findings of this study showed that on the 1st and 7th day of the rotarod test, there was no significant difference between the other study groups (P>0.05), while on the 14th day, the AlCl3 + Niosome curcumin group showed a significant difference compared to the AlCl3 group (P<0.05). In the evaluation of oxidative stress parameters, it was observed that the SOD index in AlCl3 + Niosome curcumin group showed a significant difference compared to other study groups (P<0.001). But the comparison between the groups did not show any significant difference (P>0.05). Also, the MDA index of the AlCl3 + Niosome curcumin group showed a significant difference compared to the saline group, )P<0.05(. But the comparison between the groups did not show a significant difference, )P>0.05(. In the GPX index, AlCl3 + Niosome curcumin group showed a significant difference compared to the saline group and the ethanol group, )P<0.05(. But the comparison between the groups did not show a significant difference, )P>0.05(. The TAC index of AlCl3 + Niosome curcumin group showed a significant difference compared to the AlCl3 + curcumin group, P < 0.05. But the comparison between the groups did not show a significant difference, P>0.05. The analysis of the data obtained from the changes in AMPA gene expression shows that the groups receiving curcumin, Niosome curcumin and AlCl3+ curcumin have significant difference with the ethanol group (P<0.01, P<0.01,P<0.01). There is no significant difference between other groups (P>0.05). The changes in NMDA gene expression showed that the groups receiving curcumin have a significant difference with the control group (P<0.05) (The difference is indicated by * in the figure). There is no significant difference between other groups (P>0.05). AMPA protein level. The results shows that AlCl3 and Corcomin and AlCl3 + Cur groups have significant differences with the Eth group (P<0.001). Corcomin and Niosome cur and AlCl3 +niosome cur groups have significant differences with the AlCl3 group (P<0.001). Niosome cur and AlCl3 +niosome cur and AlCl3 +Cur groups have significant differences with the Corcomin group (P<0.001). Niosome cur and AlCl3 +niosome cur groups have significant differences with the AlCl3 +Cur group (P<0.001). NMDA protein level. The results showed that AlCl3 and Niosome cur and AlCl3 + Niosome cur groups have significant differences with the control group (P<0.001). AlCl3 and Niosome cur and AlCl3 + Niosome cur groups have significant differences with the Eth group (P<0.001, P<0.01, P<0.001). Curcumin and Niosome cur and AlCl3 + Niosome cur and AlCl3 + Cur groups have significant differences with the AlCl3 group (P<0.001). Niosome cur and AlCl3 + Niosome cur groups have significant differences with Corcomin group (P<0.001). Niosome cur and AlCl3+ Niosome cur groups have significant differences with the AlCl3+ Cur group (P<0.001). The totall cells, Analysis of the results shows that AlCl3 and AlCl3 + Niosome curcumin groups only have significant difference with controll group (P<0.01, P<0.05). The curcumin group has a significant difference with AlCl3 group (P<0.001). The nisome curcumin and AlCl3 + Niosome curcumin groups have significant difference with curcumin group (P<0.05, P<0.001).Pyknotic cells. Analysis of the results shows that. The AlCl3 + curcumin and Niosome curcumin groups have significant difference with curcumin group (P<0.05, P<0.05). Relatives of pyknotic cells to total cells. the results shows that the curcumin group has a significant difference with the ethanol group (P<0.01). The curcumin group has a significant difference with AlCl3 group (P<0.001). The AlCl3 + curcumin and Niosome curcumin and AlCl3 + Niosome curcumin groups have significant difference with curcumin group (P<0.001, P<0.001, P<0.01) There is no significant difference between other groups (P>0.05). % of cerebellum weight. the results showe that all groups except the AlCl3 + Cur group have significant difference with control group (P<0.001). There is significant difference between all groups with ethanol group (10 mg/kg) (P<0.001). % of cells in cerebellum. the results showed that there is significant difference between all groups with ethanol group (P<0.001). Curcumin, AlCl3 + Cur, Niosome curcumin and AlCl3 + Niosome cur groups have significant difference with the AlCl3 group (P<0.001). AlCl3 +Cur, Niosome cur and AlCl3 +niosome cur groups have significant difference with the Curcumin group (P<0.001). Niosome cur and AlCl3 + Niosome cur groups have significant difference with the AlCl3 + Cur group (P<0.001).Conclusion: Based on the obtained results, the induction of Alzheimer's disease in rats increased the expression of AMPA and NMDA genes, and in Alzheimer's groups treated with curcumin and nisome curcumin, gene expression decreased. Also, the changes of oxidative stress parameters in the relationship between Alzheimer's induction and treatment with curcumin and curcumin nanoparticle are in line with previous researches, in such a way that the increase of MDA (unlike other oxidative stress parameters) in the Alzheimer's group and the decrease of this parameter in the treated groups with curcumin and curcumin nanoparticle, it shows the improvement of the disease. In addition, stereological and histological changes show an increase in necrosis and neuronal degeneration, hyperemia and neuroglia density and a decrease in neuron density in Alzheimer's disease, and curcumin and especially Nisome curcumin causes improvement by adjusting these parameters. In relation to the behavioral data, it was generally observed that Nisome curcumin improves Alzheimer's disease and reduces balance and movement anxiety. The resistance of rats on the Rota-Rod device is an increase in balance and movement coordination, which confirms the above statement. According to the data obtained from this research, niosomes as drug-carrying nanoparticles can be a suitable option for better performance in a shorter period of time for antioxidants such as curcumin.
