Expression Of Glycine Receptor On Endoplasmic Reticulum In Rat Cardiomyocytes

Objectives Clarify the relationship between the cytoprotective effect of glycine and the glycine receptor,and explore the co-localization relationship between glycine receptor and endoplasmic reticulum of rat’s cardiomyocytes,and the effects of hypoxia and glycine treatment on the localization of glycine receptor and endoplasmic reticulum.Methods The experiment is divided into two parts:The first Part: rat cardiomyocyte line H9C2 cells were selected.The expression of glycine receptor ɑ1(Gly R ɑ1)was detected by Western blotting under the condition of hypoxia ischemia and glycine intervention.The second part: we chose rat cardiomyocyte line H9C2 cells to observe the co-localization relationship between glycine receptor ɑ1(Gly R ɑ1)and cardiomyocyte endoplasmic reticulum by Immunofluorescence staining and confocal laser scanning microscopy.Results1)In vitro experiment: the result of effects of hypoxia and Glycine on the expression of glycine receptor ɑ1 in rat’s myocardial cells showed that hypoxia can induce an increasing expression of glycine receptor α1 in myocardial cells,while the expression of glycine receptorɑ1 was transient increase trend after the treatment of glycine.Especially,after treatment with6 h and 12 h,the expression of glycine receptor ɑ1 had a significant difference compared with0h(p<0.05),and glycine inhibited the increase of glycine receptor ɑ1 expression induced by hypoxia.2)Gly Rα1 was proved to be existed in the endoplasmic reticulum(ER)of myocardial cells.3)The results of the Co-localization of Glycine receptor α1(Gly R α1)and endoplasmic reticulum(ER)showed that glycine receptor ɑ1(Gly R α1)and endoplasmic reticulum were colocalized.And in hypoxia group(Hyp group),the expression of glycine receptor α1(Gly Rɑ1)was higher than other groups,while the expression of Gly R ɑ1 in glycine group(Group Gly)was weak.After Glycine treated on the hypoxia treatment stimulated cells,the higher expression of glycine receptor α1(Gly Rα1)that induced by hypoxia was obviously inhibited by glycine.ConclusionsThe results of this study showed that hypoxia can promote the expression of glycine receptor in myocardial cells of rats increased significantly,and the combination of glycine and glycine receptor could inhibit the higher expression of glycine.Meanwhile,Glycine can effectively reduce myocardial apoptosis rate.The co-localization study of Glycine receptorα1(Gly Rα1)and endoplasmic reticulum(ER)of rats’ myocardial cells shown that Gly Rα1was located in ER membrane,and the expression of Gly Rα1 after hypoxia treated 6h was significantly increased,which indicated that the glycine protected cells through the combination of endoplasmic reticulum Gly Rα1.However the mechanisms needs further investigation.