Effects of Exogenous Melatonin on Colon Glial Cells in Experimental Desynchronosis

E. M. Luzikova, D. O. Sumbaev, E. K. Bakhman
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Abstract

The effect of exogenous and endogenous melatonin on GFAP and S-100 positive cells of the colon mucosa of rats kept under different lighting conditions was studied. The aim was to estimate the effect of melatonin on the structural and functional characteristics of colon glial cells in normal conditions and in experimental desynchronosis. Material and methods . The study was conducted on laboratory male Wistar rats (n=72), 2-months old, weighing 180–200 g, divided into 6 groups 12 rats in each. Group I consisted of animals treated in natural light conditions; Group II – rats treated in natural light conditions, receiving synthetic melatonin (Melaxen, Unipharm, Inc., USA) ad libitum at a concentration of 4 mg/l with drinking water; Group III – animals treated in constant darkness; Group IV – animals treated in constant darkness and received melatonin; Group V consisted of animals treated under constant lighting conditions; Group VI included rats treated in constant lighting conditions and received melatonin. Astrocytes were identified using an indirect immunohistochemical method using polyclonal antibodies to glial fibrillary acidic protein (Dako, Germany 750 µg/ml) and protein S-100 (Spring Bio Science USA 1:300). Cells were counted using a Carl Zeiss Axio Scope A1 microscope at 400 magnification and SigmaScan Pro 5 software. Statistical processing was carried out using the MS Excel and Statistica 17 programs. Results. The number of GFAP positive cells decreased in groups III and V. Conditions II and VI resulted in a decrease in the number of gliocytes. In group IV, the administration of melatonin led to an increase in the number of studied cells. A significant increase in the optical density of GFAP in colon gliocytes was observed only when rats were treated under constant lighting conditions. The number of S-100 positive cells decreased in groups III and II. Conditions V and IV led to an increase in the number of S-100 positive cells. The optical density of S-100 does not depend on photoperiod. Conclusion. S-100B and GFAP positive glial cells of the colon respond both to changes in photoperiod and to the administration of exogenous melatonin. In particular, under constant dark conditions, the number of GFAP and S-100B positive cells decreases. The obtained data can be used in the elaboration of new therapeutic approaches to the treatment of intestinal diseases.
外源性褪黑素对实验性失同步结肠胶质细胞的影响
研究了不同光照条件下外源性和内源性褪黑素对大鼠结肠黏膜GFAP和S-100阳性细胞的影响。目的是评估褪黑素对正常条件下和实验失同步状态下结肠胶质细胞结构和功能特征的影响。材料和方法。实验选用2月龄、体重180 ~ 200 g的实验室雄性Wistar大鼠72只,分为6组,每组12只。第一组为自然光条件下处理的动物;II组:在自然光条件下,给予合成褪黑素(Melaxen, Unipharm, Inc., USA),浓度为4mg /l,随饮用水随意注射;第三组——在持续黑暗中治疗的动物;IV组:持续黑暗并接受褪黑素治疗;V组是在恒定光照条件下处理的动物;第六组包括在恒定光照条件下接受褪黑激素治疗的大鼠。使用胶质纤维酸性蛋白(Dako, Germany, 750µg/ml)和S-100蛋白(Spring Bio Science USA, 1:30 00)的多克隆抗体,采用间接免疫组化方法鉴定星形胶质细胞。使用400倍率的卡尔蔡司Axio Scope A1显微镜和SigmaScan Pro 5软件进行细胞计数。采用MS Excel和Statistica 17软件进行统计处理。结果。III组和v组GFAP阳性细胞数量减少。条件II和VI导致胶质细胞数量减少。在第四组中,褪黑激素的使用导致研究细胞数量的增加。只有当大鼠在恒定光照条件下处理时,才观察到结肠胶质细胞中GFAP的光密度显著增加。ⅲ组和ⅱ组S-100阳性细胞数量减少。条件V和条件IV导致S-100阳性细胞数量增加。S-100的光密度与光周期无关。结论。结肠的S-100B和GFAP阳性胶质细胞对光周期的变化和外源性褪黑激素的管理都有反应。特别是在恒定黑暗条件下,GFAP和S-100B阳性细胞数量减少。所获得的数据可用于制定治疗肠道疾病的新治疗方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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