健脾益气方含药血清下调Vimentin蛋白水平对人肝癌细胞SMMC-7721的影响  

Effects on SMMC-7721 cell behavior due to down-regulation of Vimentin expression by serum containing Jianpi Yiqi decoction

作　　者：卓少元[1] 谢金玲[1] 

机构地区：[1]广西中医药大学基础医学院,广西南宁530200

出　　处：《时珍国医国药》2017年第1期55-59,共5页Lishizhen Medicine and Materia Medica Research

基　　金：广西自然科学基金青年基金(No.2014GXNSFBA118145)

摘　　要：目的研究健脾益气方下调Vimentin蛋白水平对肝癌细胞的影响,探讨健脾益气方防治肝癌的机制。方法利用TGF-β1诱导的人肝癌细胞SMMC-7721 EMT模型,采用血清药理学方法,并结合Vimentin裂解剂和Caspase抑制剂,观察7.5%、15%、30%浓度健脾益气方含药血清干预对肝癌细胞增殖、侵袭、凋亡,及Vimentin、Caspase-3、PARP-1与Caspase-1蛋白表达的影响。结果与空白血清组比较,健脾中、高剂量组健脾益气方含药血清均可以降低肝癌细胞Vimentin和PARP-1蛋白表达量(P<0.01),其下调程度与二者的裂解程度,及肝癌细胞的增殖抑制率、侵袭抑制率和凋亡率趋势一致;并升高肝癌细胞Caspase-3、降低Caspase-1蛋白的表达(P<0.01)。Vim裂解组细胞的增殖抑制率、侵袭抑制率和早期凋亡率在各实验组中均为最高值,其肝癌细胞中各蛋白表达趋势与健脾组一致,但下调或上调程度更明显。Casp抑制组细胞的增殖抑制率、侵袭抑制率在各实验组均为最低值,早期凋亡率在各实验组中也仅高于EMT模型组,两组间差异无统计学意义(P>0.05);细胞中Caspase-3蛋白表达下调(P<0.01),PARP-1表达上调(P<0.01),Vimentin表达差异无统计学意义(P>0.05),Vimentin和PARP-1无蛋白裂解片段出现。健脾各剂量组中以中剂量组效果最佳。结论 15%浓度的健脾益气方含药血清对人肝癌细胞SMMC-7721的干预效果最佳。健脾益气方可能通过下调人肝癌细胞SMMC-7721中Vimentin蛋白的表达,抑制肝癌细胞的增殖与侵袭、并诱导部分肝癌细胞凋亡;导致Vimentin蛋白下调的原因可能在于Caspase-3对它的裂解。此过程中,健脾益气方一方面利用Caspase-3/Vimentin裂解片段形成的正反馈促凋亡信号诱导肝癌细胞发生凋亡,另一方面可能利用Vimentin水平的下调导致NLRP3/Caspase-1炎症信号通路减弱,干预肝癌炎症微环境。Objective Toinvestigate the effects on SMMC-7721 cell behavior due to down-regulation of Vimentin expression by serum containing Jianpi Yiqi decoction. Methods The effects on biological behavior of TGF-β1-induced SMMC-7721 were examined after treatment by 7. 5%,15%,30% concentration medicated serum containing Jianpi Yiqidecoction,and compared with the results by Vimentin breaker or by Caspase inhibitor. Results Compared with the blank serum group,decreased protein expression of Vimentin,PARP-1 and Caspase-1（ P 〈 0. 01）,and increased protein expression of Caspase-3 were observed. The decreased degree of Vimentin and PARP-1 were consistent with the proliferation and invasion inhibition rate and early apoptosis rate of SMMC-7721 cells. The proliferation and invasion inhibition rate and early apoptosis rate of the Vimentin-breaker group cells were the highest in all experimental groups,but only up-regulation or down-regulation of the proteins expressionis more obvious. However,the proliferation and invasion inhibition rate of the Caspase-inhibitor group cells were the lowest in all experimental groups,and the early apoptosis rate was only higher than the EMT model group（ P 〈 0. 05）. Compared with the blank serum group cells,down-regulation of Caspase-3 and up-regulation of PARP-1protein expression were observed in the Caspase-inhibitor group cells. Vimentin expression difference was not statistically significant（ P 〉 0. 05）,Vimentin and PARP-1 show no protein cleavage appear. Conclusion The expression of Vimentin in SMMC-7721 cells was down-regulated by serum containing Jianpi Yiqi decoction,especially 15% serum concentration medicated serum,which may be due to Vimentin cleavage by Caspase-3. In this process,a positive feedback loop forms,whereby activated caspase-3degrade Vimentin,and these degraded fragments consequentlyactivate caspases to amplify apoptosis. Furthermore,down-regulation of Vimentin may be resulted indecreased NLRP3/Caspase-1 inflammatory signaling pathway and regulate liver infla

关 键 词：健脾益气方 SMMC-7721 VIMENTIN Caspase-3 CASPASE-1 

分 类 号：R285.5[医药卫生—中药学]