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The Effect of Hyperglycemia on the Expression of Stress Proteins of Endoplasmic Reticulum and Vascular Smooth Muscle Cell (VSMC) Migration from Normal Rats
Author(s) -
MejíaElizondo Rebeca,
HernándezMéndez Aurelio,
ChiAhumada Erika,
EspinosaTanguma Ricardo
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.1017.5
Subject(s) - endoplasmic reticulum , unfolded protein response , vascular smooth muscle , microbiology and biotechnology , glucose regulated protein , cell , cell migration , myosin , biology , medicine , endocrinology , chemistry , biochemistry , smooth muscle
Diabetes mellitus (DM) is a chronic degenerative disease characterized by high glucose levels on blood, which is the leading cause of micro and macrovascular changes observed in this disease. At cellular level DM promotes the misfolding of proteins in the endoplasmic reticulum (ER), a condition defined as ER stress. ER stress is characterized by an increase on the expression of some proteins, unfolded protein response (UPR), that promotes the decrease of the unfolded proteins translation and the increase in the expression of folding proteins or apoptosis when the damage is too great. Since research efforts on macrovascular disease secondary to DM are focused on pathological changes on endothelial cells, we are interested in evaluating the changes that occur at the level of VSMC as it has been demonstrated that they play a fundamental role in the pathophysiological process. On this research we obtained primary smooth muscle cell cultures from the thoracic aorta of normal rats by means of explant culture technique. The cells were characterized by immunofluorescence against muscle alpha‐actin and myosin and they were used on experiments were the cells were exposed to high glucose concentrations (15 mM) where we analyzed: proteins related to endoplasmic reticulum stress, expression of Ca +2 ATPase of the ER (SERCA2), cellular migration through the method of wound healing assay and the usage of the Boyden camera. Our results show that muscle cells exposed to 15 mM of glucose for 48 hours do not exhibit any changes on cell migration or in the expression of ATF‐6 and XBP‐1 proteins (analyzed by western blot), that we used as stress markers. However cells exposed for one week to 15 mM glucose showed an increase on cell migration as well as an increase on the expression of stress markers, which leads us to believe that these cells have a phenotype that is similar to those from diabetic rats. On the other hand the expression of SERCA2, analyzed by inmunoblot, seems to increase which suggests that the cells that are exposed to hyperglycemia over express SERCA2 to compensate for the damage on the ER being that the activity of this ATPase is fundamental to maintain a proper cell function. Overall these results show that the expression of smooth muscle cells to high glucose concentrations simulates the process that takes place in cells from diabetic animals, which allows us to have a study model of diabetes without inducing animals with drugs that produce severe side effects. Support or Funding Information Supported by Conacyt to RET: 223350