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AIM: To establish the endoplasmic reticulum stress( ERS) cell model in vascular smooth muscle cells( VSMCs) of SpragueDawley( SD) rats. METHODS: Under sterile condition,the coronary arteries were isolated from SD rats. The primary VSMCs were cultured by tissue-sticking method,and observed the basic morphological characteristics under optical microscope. The marker proteins of VSMCs including α-smooth muscle actin( α-SMA) and smooth muscle myosin heavy chain( SM-MHC) were identified by immunofluorescence technique. VSMCs were treated with thapsigargin(0. 5,1 and 2 μmol / L) for 24 h,and the expression levels of binding immunoglobulin protein( Bi P) and C / EBP homologus protein( CHOP),the marker molecules of ERS,were detected using Western blotting. RESULTS: VSMCs climbed out from coronary artery tissues after about six days,and the cells had a nice state and formed the VSMC-like typical “ peak valley”. The results of immunofluorescence technique show that the marker proteins of VSMCs,α-SMA and SM-MHC were expressed significantly. The results of Western blotting show that the protein expression levels of Bi P and CHOP were increased by thapsigargin in a dose-dependent manner. CONCLUSION: VSMCs can be successfully cultured by tissue-sticking method and built the ERS model induced by thapsigargin.
AIM: To establish the endoplasmic reticulum stress (ERS) cell model in vascular smooth muscle cells (VSMCs) of Sprague Dawley (SD) rats. METHODS: Under sterile conditions, the coronary arteries were isolated from SD rats. The primary VSMCs were cultured by tissue -sticking method, and observed the basic morphological characteristics of optical microscope. The marker proteins of VSMCs including α-smooth muscle actin (α-SMA) and smooth muscle myosin heavy chain (SM-MHC) were identified by immunofluorescence technique. with thapsigargin (0.5,1 and 2 μmol / L) for 24 h, and the expression levels of binding immunoglobulin protein (Bi P) and C / EBP homologus protein (CHOP), the marker molecules of ERS, were detected using Western blotting. RESULTS: VSMC-like typical “peak valley”. The results of immunofluorescence technique show that the marker proteins of VS The results of Western blotting show that the protein expression levels of Bi P and CHOP were increased by thapsigargin in a dose-dependent manner. CONCLUSION: VSMCs can be successfully cultured by tissue- sticking method and built the ERS model induced by thapsigargin.