Effects of BAPTA-AM on acid-induced autophagy of rat articular chondrocytes and its possible mechanisms
10.3969/j.issn.1001-1978.2015.05.013
- VernacularTitle:钙络合剂BAPTA-AM对胞外酸化诱导大鼠关节软骨细胞自噬作用的影响及其可能机制
- Author:
Wenfan GAO
;
Feihu CHEN
;
Jinfang GE
;
Ziyun DENG
;
Jing LEI
;
Renpeng ZHOU
;
Zhisen WANG
- Publication Type:Journal Article
- Keywords:
BAPTA-AM;
rheumatoid arthritis;
chon-drocytes;
autophagy;
Ca2+;
extracellular acidification
- From:
Chinese Pharmacological Bulletin
2015;(5):655-659
- CountryChina
- Language:Chinese
-
Abstract:
Aim To observe the effect of BAPTA-AM on extracellular acid-induced autophagy in rat articular chondrocytes and its possible mechanisms.Methods Rat articular chondrocytes were isolated from Sprague-Dawley rats and incubated with different pH medium. The states of autophagy were examined by acridine or-ange (AO ) staining .Moreover,the expressions of LC3 ,Beclin-1 ,ULK1 ,CaMKKβ,AMPK and mTOR were detected using Western blot or quantitative real-time PCR (qRT-PCR ). Intracellular calcium ([Ca2+]i )was analyzed by a Ca2+-imaging method. Results Compared with pH 6.0 group,BAPTA-AM could significantly decrease the activation of autophagyinduced by acid exposure,and the expressions of autophagy markers including LC3 Ⅱ,Beclin1 and ULK1were also decreased,accompanied with reduced acidinduced [Ca2 +]i influx,decreased proteins expressionof CaMKKβand phosphorylatedAMPK,and increasedphosphorylation of mTOR.Conclusion BAPTAAMcan significantly restrain acidinduced autophagy in ratarticular chondrocytes,the mechanism of which may beassociated with decreased Ca2 + influx.
