Model construction of rat coronary artery smooth muscle cell endoplas-mic reticulum stress induced by thapsigargin
10.3969/j.issn.1000-4718.2017.01.021
- VernacularTitle:毒胡萝卜素诱导大鼠冠状动脉平滑肌细胞内质网应激模型的建立
- Author:
Xiaoyan CHEN
;
Chunyu DENG
;
Sujuan KUANG
;
Hui YANG
;
Fang RAO
;
Zhixin SHAN
;
Qiuxiong LIN
;
Li JIANG
- Keywords:
Coronary artery smooth muscle cells;
Tissue explant method;
Endoplasmic reticulum stress
- From:
Chinese Journal of Pathophysiology
2017;33(1):128-132
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress ( ERS) model in CASMCs of SD rats.METHODS:CASMCs were cultured by tissue explant method .The morphological characteristics were observed under optical micro-scope.The marker proteins of CASMCs , including α-SMA and SM-MHC, were identified by immunofluorescence tech-nique.The protein expression levels of BiP and CHOP , the marker molecules of ERS, were determined by Western blot . RESULTS:The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical hill and valleygrowth pattern of CASMCs at 9~10 d.The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed .The results of Western blot showed that the protein expression of BiP and CHOP in TG ( 1 and 2 μmol/L ) treatment groups was increased compared with control group .Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION:CASMCs can be successfully cultured by tissue explant method .ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.
