siRNA沉默β-arrestin2促进肝星状细胞凋亡  

Small interfering RNA targeting β-arrestin2 promoted apoptosis of hepatic stellate cells

作　　者：宋杨[1] 孙妩弋[1] 胡姗姗[1] 汪庆童[1] 魏伟[1] 

机构地区：[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

出　　处：《中国药理学通报》2012年第5期612-616,共5页Chinese Pharmacological Bulletin

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)

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。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.

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 

分 类 号：R-332[医药卫生] R322.47