[1]高文凡,陈飞虎,葛金芳,等.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[J].中国药理学通报,2015,(05):655-659.
 GAO Wen-fan,CHEN Fei-hu,GE Jin-fang,et al.Effects of BAPTA-AM on acid-induced autophagy of rat articular chondrocytes and its possible mechanisms[J].Chinese Pharmacological Bulletin,2015,(05):655-659.
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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()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2015年05期
页码:
655-659
栏目:
论 著
出版日期:
2015-05-15

文章信息/Info

Title:
Effects of BAPTA-AM on acid-induced autophagy of rat articular chondrocytes and its possible mechanisms
作者:
高文凡陈飞虎葛金芳邓子云雷 静周仁鹏王志森
安徽医科大学药学院,安徽 合肥 230032
Author(s):
GAO Wen-fan CHEN Fei-huGE Jin-fang DENG Zi-yun LEI Jing ZHOU Ren-peng WANG Zhi-sen
School of Pharmacy, Anhui Medical University, Hefei 230032,China
关键词:
BAPTA-AM 类风湿关节炎 关节软骨细胞 细胞自噬 Ca2+ 胞外酸化
Keywords:
BAPTA-AM rheumatoid arthritis chondrocytes autophagy Ca2+ extracellular acidification
分类号:
R-332; R322.72; R329.24; R394.2; R977.3
文献标志码:
A
摘要:
目的 观察 BAPTA-AM 对胞外酸化诱导大鼠关节软骨细胞自噬作用的影响,探讨其可能的作用机制。方法 体外使用胰酶-Ⅱ型胶原酶消化法分离提取大鼠关节软骨细胞,分为正常组(pH 7.4)、酸化组(pH 6.0)、以及分别经 BAPTA-AM 处理的正常组和酸化组,激光共聚焦技术检测软骨细胞胞内钙离子变化,实时荧光定量 PCR 法检测细胞自噬基因 Beclin-1、ULK1 mRNA 的表达,Western blot 法检测自噬蛋白 LC3 的表达,吖啶橙染色分析细胞自噬溶酶体形成情况。结果 与 pH 7.4 正常组比较,pH 6.0 酸化刺激明显增加大鼠关节软骨细胞内 Ca2+的浓度,且自噬标志物 Beclin-1、ULK1 mRNA 及 LC3Ⅱ 蛋白表达均明显升高,酸性自噬溶酶体形成增多,同时酸化刺激能引起 CaMKKβ 及 p-AMPK 蛋白表达水平增高,磷酸化蛋白 p-mTOR 水平明显降低。BAPTA-AM 酸化组自噬水平和 CaMKKβ 及 p-AMPK 表达明显降低,p-mTOR 表达明显升高。结论 BAPTA-AM 能明显减弱胞外酸化诱导软骨细胞自噬作用,其机制可能与抑制胞内Ca2+有关。
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 orange(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 autophagy induced by acid exposure, and the expressions of autophagy markers including LC3Ⅱ, Beclin-1 and ULK1 were also decreased, accompanied with reduced acid-induced [Ca2+]i influx, decreased proteins expression of CaMKKβ and phosphorylated-AMPK, and increased phosphorylation of mTOR. Conclusion BAPTA-AM can significantly restrain acid-induced autophagy in rat articular chondrocytes, the mechanism of which may be associated with decreased Ca2+ influx.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2015-01-29,修回日期:2015-03-11 基金项目:国家自然科学基金资助项目(No 81271949) 作者简介:高文凡(1991-),女,硕士生,研究方向:分子药理学,E-mail:gaowenfan0829@163.com; 陈飞虎(1962-),男,博士,教授,博士生导师,研究方向:抗炎免疫药理学、分子药理学,通讯作者,E-mail: cfhchina@sohu.com
更新日期/Last Update: 1900-01-01