Method of Housing and Transfer and Experimental Autoimmune Encephalomyelitis: An Experimental Study on C57BL/6

Background: Multiple Sclerosis (MS) is an autoimmune inflammatory demyelinating disease of human central nervous system. Although experimental autoimmune encephalomyelitis (EAE) is the most commonly used method to induce MS, there are unexpected results in the modeling outcomes, which led to inappropriate clinical score scaling. Recent studies focused on the possible factors that may affect the final outcome of EAE modeling. Some of these factors were observed and discussed in our experiment on C57BL/6 model. Objectives: The present research was carried out to find the possible effects of environmental factors, including transfer, handling, housing, and dark-light cycle on EAE modeling scoring. Materials and Methods: Twenty female mice (C57BL/6) were used that divided into two groups (n = 10) by random. The routine method of MS induction in mammals was used in both groups. Following induction, animals of group one were placed in a separated room with the least local translocation and handling, whereas animals of the second group were placed in the same room as the other animals with normal local allocation as others. The animals were observed and scored using routine clinical scoring for EAE. Results: Our data showed that the EAE induction in group one was significantly more successful than group two (with the mean score > 3). Conclusions: Although the EAE is still a scientific method to induce MS in rodents, it requires more attention to environmental factors that might influence the result. The mechanisms of these factors are unknown, but it seems that the role of housing environment should be taken into consideration

Effects of Exogenous Estrogen Treatment on Hippocampal Neurogenesis of Diabetic Ovariectomized Rats

Background: Mellitus Diabetes (DM) is the most important metabolic diseases. The incidence of DM is prone to increase. Vasculopathy, retinopathy, central and peripheral neuropathy are the most important reported side effects of DM. Cognitive dysfunction following DM reported in both sexes. Hippocampus is a major part of brain involving in cognitive function, its cells are able to neurogenesis, so it is possible that DM affects the hippocampus. In addition, neuroprotective effects of female sex steroids are reported elsewhere. In order to answer the question of whether female sex steroid are able to suppress the effects of DM on neurogenesis of dentate gyrus (DG) in diabetic ovariectomized rat the present study designed.Methods: Sprague-Dawley adult female rats were used in this study. The animals randomly divided in 8 groups including; control, diabetic (Diab), ovariectomy (OVX), Diab+OVX, estrogen treated (E2; Diab+OVX+E2), surgical and vehicle sham. Intrapritoneal injection of STZ, subcutaneous injection of E2 and routine bilateral surgery were used respectively to induce diabetes, estrogen treatment and OVX. Nissl staining, Brdu immunohistochemistry (IHC) and western blotting were used in this study. Statistical analysis was done and the results presented in mean ± SD, Pv < 0.05 considered significant.Results: Brdu IHC showed that the neurogenesis significantly decreased in OVX, Diab and OVX-Diab groups (Pv < 0.05) in comparison with control and sham groups. Western blotting showed significant increase of Bax and decrease of Bcl2 proteins of trial groups comparing to control. Estrogen treatment significantly improved neurogenesis in animals of Diab+OVX+E2 group. The neurogenesis impairment was more sever in OVX + Diab animals than OVX and Diab ones merely.Conclusion: Based on our data, cognitive dysfunction caused by DM is related to hippocampal neurogenesis reduction and might improve under the influence of ovarian steroidal hormone therapy

Effects of 660nm Low-Level Laser Therapy on P2X3 Expression of Lumbar DRG of Adult Male Rats with Neuropathic Pain

Background: Neuropathic Pain (NP) is a serious suffering medical condition that frequently leads to disability and life style changes. Although the exact mechanisms of NP are still unknown, recently the role of reactive oxygen species (ROS) reported as an important factor for NP. Apoptosis, increase of ATP production and reduction of antioxidants are also the other factors influencing in NP. There are certain therapeutic procedures for NP among them using laser therapy newly received more attention. In the present research we studied the molecular effects of Low Level Laser Therapy (LLLT) on a rat model of NP.Methods: Thirty adult male Wistar rats (200-250 g) that randomly divided into three groups including chronic constriction injury (CCI), CCI+LLLT and control were used in this study. CCI technique was used to induce NP. Laser therapy was done by using laser beam of 660 for 14 days following CCI. After that, expression of P2X3 of the DRG, Bax and Bcl2 in lumbar spinal segments measured by Western Blotting. Level of glutathione (GSH) was also measured in lumbar spinal cord segments by Continuous Spectrophotometric Rate Determination method. For behavioral study the mechanical and thermal hyperalgesia were evaluated in days 7 and 14 after CCI.Results: LLLT for two weeks increased expression of Bcl2 and GSH, whereas decreased Bax and P2X3 expression significantly. Comparing the results of behavioral study showed significant differences in the mechanical and thermal threshold showed between CCI and CCI+ LLLT groups.Conclusion: Based on our findings, the therapeutic effects of LLLT for NP act throughout cellular and molecular mechanisms which improve mitochondrial function that in turn improve cell function and prevent apoptosis.