酸敏感离子通道阻断剂阿米洛利对佐剂性关节炎大鼠关节软骨细胞凋亡抑制作用的初步研究

中国临床药理学与治疗学 ›› 2010, Vol. 15 ›› Issue (1): 41-46.

酸敏感离子通道阻断剂阿米洛利对佐剂性关节炎大鼠关节软骨细胞凋亡抑制作用的初步研究

张腾跃¹, 陈飞虎¹, 王念¹, 袁凤来², 胡伟¹, 吴繁荣¹

¹安徽医科大学药学院, 合肥230032, 安徽;
²南通大学第三附属医院, 无锡214041, 江苏

收稿日期:2009-09-28 修回日期:2009-11-09 出版日期:2010-01-26 发布日期:2020-09-21
通讯作者: 陈飞虎, 男, 博士, 教授, 博士生导师, 研究方向:分子药理学。Tel:0551-5161115 　E-mail:cfhchina@sohu.com
作者简介:张腾跃, 男, 硕士, 助教, 研究方向: 分子药理学。Tel: 13855188692 E-mail: zhty6580@sina.com
基金资助:
国家自然科学基金资助(30873080 、30901526)

Inhibitions of articular cartilage cells apoptosis in rats with adjuvant arthritis by Amiloride, a blocker of acid-sensing ion channels

ZHANG Teng-yue¹, CHEN Fei-hu¹, WANG Nian¹, YUAN Feng-lai², HU Wei¹, WU Fanrong¹

¹School of Pharmacy, Anhui Medical University, Hefei 230032, Anhui, China;
²The Third Affiliated Hospital of Nantong University, Wuxi 214041, Jiangsu, China

Received:2009-09-28 Revised:2009-11-09 Online:2010-01-26 Published:2020-09-21

摘要/Abstract

摘要： 目的研究酸敏感离子通道阻断剂阿米洛利(Amilo ride) 对佐剂性关节炎(AIA) 大鼠关节软骨组织的保护作用及机制。方法将大鼠随机分为正常组、AIA 模型组、阿米洛利(2 .5 、5 .0 、10 .0 mg·kg-1·d-1) 组和阿司匹林(50 mg/kg)对照组。弗氏完全佐剂(CFA) 致AIA 后第10天, 大鼠出现继发性炎症, 此时阿米洛利、阿司匹林灌胃给药, 连续8 d ;正常组与模型组给予等容量的无菌注射用水。检测大鼠继发侧关节肿胀度, 光学显微镜观察关节病理变化, 透射电镜、Tunel 法观察阿米洛利对AIA 大鼠关节软骨细胞凋亡的影响, 免疫组织化学技术测定阿米洛利对AIA 大鼠关节软骨中的Ⅱ型胶原(COII) 蛋白合成的影响, Alcian 染色测定阿米洛利对AIA 大鼠关节软骨中的PG 合成的影响。结果阿米洛利各剂量组对A IA 大鼠的足肿胀未见明显影响;病理学观察关节软骨表面光滑, 软骨细胞层次尚清晰, 成熟软骨细胞数量增加;透射电镜观察软骨细胞核内异染色质轻度边集、细胞内粗面内质网轻度扩张、线粒体轻度肿胀, 与模型组软骨细胞核膜皱缩、核染色质高度边集、核膜破裂、出现凋亡小体相比无明显细胞凋亡的特征;Tunel法原位检测显示凋亡阳性细胞数显著减少(P <0 .01) ;免疫组化及Alcian 染色检测显示关节软骨基质成分COII 蛋白和PG 的表达量增加。结论 ASICs 阻断剂阿米洛利可通过抑制AIA 大鼠关节软骨细胞凋亡发挥对关节软骨的保护作用。

关键词: 阿米洛利(Amilo ride), 佐剂性关节炎(AIA), 酸敏感离子通道(ASICs), 细胞凋亡

Abstract: AIM: To investig ate the protective effects of articular cartilage by acid-sensing ion channels (ASICs) blocker A miloride and its mechanisms.METHODS: The rats were randomly divided into normal group, AIA model group, Amiloride (2.5, 5.0, 10.0 mg·kg -1·d-1) group, aspirin (50 mg/kg) group.The AIA model rats were induced by CFA.The secondary inflammat ion of AIA rats was appeared on the tenth day via intradermal injection CFA.The drug groups were given corresponding experimental drugs, the model group and the normal group were administered with the corresponding capacity of sterile water, lasting for eight day s.The right hind paw articular swelling degree was determined with volume meter, the morphological changes of articular cartilage was observed by light microscope.The apoptosis of articular cartilage cells were observed by transmission electron microscope, Tunel method. The typeII collagen (COII) expressions of extracellular matrix of articular cartilage were determined by using immunohistochemistry.The proteoglycan (PG) contents of the cartilaget issue were detected by Alcian blue staining.RESULTS: Amiloride have no conspicuous effect on the hind paw swelling with AIA rats.The pathology ical study showed that the surface of articular cartilage was slick, the layer was lampros and the quantity of chondrocyte was incresaed.Transmission electron micro scope study showed that condensation and crack of nucleus and apoptotic bodies were appeared in articular cartilage cells in model group, and the condensation of heterochromatin, the distention of rough endoplasmic reticulum and the swell of bioblast were relieved in Amiloride groups.The apoptosis cell calculation in the experiment groups were fewerthan those in the model group (P <0.01) by Tunel method.The COII and PG of extracellular matrix of articular cartilage were significantly increased in Amiloride groups by immunohistochemical method and Alcian blue staining.CONCLUSION: ASICs blocker Ami loride may have protective effects on articular cartilage by inhibiting the apoptosis of articular cartilage cells of adjuvant-induced arthritis rats.

Key words: Amiloride, Adjuvant-induced arthritis(AIA), Acid sensing ion channels(ASICs), Apo ptosis

中图分类号:

R965.2

张腾跃, 陈飞虎, 王念, 袁凤来, 胡伟, 吴繁荣. 酸敏感离子通道阻断剂阿米洛利对佐剂性关节炎大鼠关节软骨细胞凋亡抑制作用的初步研究[J]. 中国临床药理学与治疗学, 2010, 15(1): 41-46.

ZHANG Teng-yue, CHEN Fei-hu, WANG Nian, YUAN Feng-lai, HU Wei, WU Fanrong. Inhibitions of articular cartilage cells apoptosis in rats with adjuvant arthritis by Amiloride, a blocker of acid-sensing ion channels[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(1): 41-46.

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