摘要
目的:探讨酸敏感离子通道1a(ASIC1a)在酸诱导的大鼠肝星状细胞活化中的作用。方法:体外培养肝星状细胞株(HSC-T6)经处理后,乳酸脱氢酶(LDH)检测细胞毒性变化,RT-PCR检测ASIC1a mRNA表达,Western Blot和RTPCR分别检测Calpain、Calcineurin蛋白和mRNA表达变化,免疫细胞化学法检测α-平滑肌肌动蛋白(α-SMA)的变化。结果:细胞酸化处理后ASIC1a含量明显上升;随着胞外pH值的下降,LDH释放量逐渐升高,ASIC1a阻滞剂可抑制酸化诱导的肝星状细胞LDH的释放量;酸化组肝星状细胞Calpain、Calcineurin mRNA和蛋白表达水平均明显增高,ASIC1a阻断剂组Calpain、Calcineurin mRNA和蛋白表达水平较酸化组明显降低;与酸化组相比,阻断ASIC1a可以降低HSC-T6中α-SMA的表达。结论:胞外酸化环境下可诱导肝星状细胞活化,阻断ASIC1a能明显降低酸化诱导的细胞活化。
AIM： To investigate the role of acid-sensing ion channel 1a in acid-induced activation of Hepatic stellate cells. METHODS： HSC-T6 cells were incubated in different pH medium without or with amiloride or psalmotoxin1 for 3 hours, which were non-specific or specific blockers of ASIC1α, respectively. Cell cytotoxicity was measured by Lactate Dehydrogenase Release Assay Kit and the mRNA expressions of ASIC1α was detected through reverse transcription polymerase chain reaction （RTPCR）. The mRNA and protein expression of Calpain and Calcineurin in different group were detected by RT-PCR or Western Blot. RESULTS： The cytotoxicity of the HSC-T6 cells was increased by extracellular acidosis with both pH 6.5, pH 6.0 and pH 5.5 condition, and the cytotoxicity was decreased by non-specific andspecific blockers of ASIC1α. As the pH declined, the mRNA expression of ASIC1α increased gradually. Compared with that incubated in pH 7.4 medium, the protein and mRNA expression of Calpain and Calcineurin in HSCs with acidification medium （pH 6.0） was significantly increased. Amiloride and PcTx-1 could both remarkably decrease the expression of Calpain and Calcineurin. ASICla inhibitors can also reduce the expression of α-SMA. CONCLUSION. These results demonstrated that ASICla is involved in acid-induced activation of hepatic stellate cells.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2014年第5期528-533,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家自然科学基金(81100301)
安徽医科大学七年制早期接触科研训练计划(2013-ZQKY-63)
关键词
酸敏感离子通道
肝星状细胞
活化
acid-sensing ion channel 1a hepatic stellate cells
activation
