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siRNA沉默β-arrestin2促进肝星状细胞凋亡

ISSN号：1001-1978
期刊名称：Chinese Pharmacological Bulletin
时间：2012
页码：612-616
分类：R-332[医药卫生] R322.47[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
作者机构：[1]安徽医科大学临床药理研究所,抗炎免疫药理学教育部重点实验室,抗炎免疫药物安徽省工程技术研究中心,安徽合肥230032
相关基金：国家自然科学基金资助项目（No 30973543;81173075）; 高等学校博士学科点专项科研基金项目（No 20113420120002）; 安徽高校省级自然科学研究项目（No KJ2012A153）; 安徽省自然科学基金项目（No 090413108）; 安徽省高等学校省级优秀青年人才基金项目（No 2010SQRL079）
相关项目：B细胞刺激因子受体BAFF-R、BCMA和TACI介导信号和相互关系在胶原性关节炎发病中作用及受体抑制剂对其的影响

关键词：肝星状细胞, 小分子干扰RNA, β-arrestin2, 凋亡, BCL-2, BAX, 肝纤维化, hepatic stellate cells, small interfering RNA, β-arrestin2, apoptosis, Bcl-2, Bax, hepatic fibrosis

中文摘要：

目的研究小干扰RNA（small interfering RNA,siRNA）干扰β-arrestin2表达对大鼠肝星状细胞株（hepatic stellatecell T6,HSC-T6）凋亡的影响。方法将化学合成的siRNAβ-arrestin2以Lipofectamine包裹,转染HSC-T6细胞,设阴性对照和空白对照;采用逆转录-聚合酶链反应和Westernblot检测细胞β-arrestin2表达;采用Western blot检测细胞Bcl-2、Bax的表达;采用流式细胞仪检测细胞凋亡。结果转染siRNA的HSC-T6细胞β-arrestin2基因及蛋白表达水平明显下调,β-arrestin2 mRNA水平和蛋白表达水平比对照组分别下调了70%±1.76%和68.43%±2.88%（P〈0.01）;Bcl-2蛋白被抑制了32.58%±3.46%（P〈0.01）,而Bax表达增加38.00%±3.72%（P〈0.01）;转染β-arrestin2 siRNA的HSC-T6细胞凋亡率为37.5%,明显高于正常HSC对照组。结论 siRNAβ-arrestin2能高效抑制HSC-T6细胞β-ar-restin2表达明显减少Bcl-2的表达,增加Bax的表达,从而促进HSC-T6凋亡,具有预防及治疗肝纤维化的潜力。

英文摘要：

Aim To investigate the effect of synthetic small interfering RNA（siRNA）targeting β-arrestin2 on the apoptosis of hepatic stellate cells（HSC）in vitro.Methods Synthetic siRNA targeting β-arrestin2 was transfected into HSC-T6 cells by lipofectamine package.Negative siRNA transfection and no transfection were used as negative and blank control,respectively.After incubation with siRNA,total RNA and protein of HSC-T6 cells were extracted.The expression of β-arrestin2 gene and protein was detected by reverse transcription-polymerase chain reaction（RT-PCR） and Western blotting.HSC-T6 apoptosis was measured by flow cytometry.The expression of Bcl-2 and Bax were determined by Western blot.Results After HSC-T6 cells were transfected with β-arrestin2 siRNA,the level of β-arrestin2 mRNA and protein expression was decresed by 70%±1.76%（P0.01）and 68.43%±2.88%（P0.01） as compared with the control group.Bcl-2 expression was also inhibited by 32.58%±3.46%（P0.01）,while Bax expression was increased by 38.00%±3.72%（P0.01）.The apoptosis rate of HSC-T6 transfected with β-arrestin2 siRNA was 37.5%,which was significantly higher than control.Conclusion Inhibition of β-arrestin2 by siRNA may have a potential effect on prevention and treatment of hepatic fibrosis by promoting apoptosis of HSC.