论文部分内容阅读
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 To investigate the effects of “Jianpi Yiqi” on down-regulating the protein level of Vimentin in hepatocarcinoma cells and to explore the mechanism of “Jianpi Yiqi Fang” in preventing and treating hepatocellular carcinoma. Methods The human hepatocellular carcinoma SMMC-7721 EMT model induced by TGF-β1 was used to observe the effects of Jianpi Yiqi Recipe containing 7.5%, 15% and 30% concentration of serum on the EMT model with serum pharmacology and Vimentin cleavage and Caspase inhibitors Effects of Intervention on Proliferation, Invasion, Apoptosis and Expression of Vimentin, Caspase-3, PARP-1 and Caspase-1 in Hepatocellular Carcinoma Cells. Results Compared with the blank serum group, the drug-containing serum of Jianpi Yiqi Recipe could decrease the protein expression of Vimentin and PARP-1 (P <0.01), and the degree of down-regulation and the degree of cleavage (P <0.01). The inhibitory rates of invasion, invasion and apoptosis of hepatocellular carcinoma cells were the same. The expressions of Caspase-3 and Caspase-1 in hepatocellular carcinoma cells were also increased. The proliferation inhibition rate, invasion inhibition rate and early apoptosis rate of Vim lysis group cells were the highest in each experimental group, and the expression trend of all the proteins in HCC cells was the same as that in Jianpi group, but the level of down-regulation or up-regulation was more obvious. The inhibitory rate of proliferation and invasion inhibition of Casp inhibition group were the lowest in all experimental groups, and the early apoptosis rate was also higher in EMT model group than in EMT model group (P> 0.05 ). The protein expression of Caspase-3 in cells was down-regulated (P <0.01), PARP-1 was up-regulated (P <0.01), Vimentin expression was not significantly different (P> 0.05) . Spleen in each dose group to the best effect in the dose group. Conclusion The interventional effect of 15% concentration Jianpi Yiqi recipe-containing serum on human hepatoma SMMC-7721 cells is the best. Jianpi Yiqi Prescription may down-regulate the expression of Vimentin in human hepatocellular carcinoma cell SMMC-7721, inhibit the proliferation and invasion of hepatocellular carcinoma cells and induce the apoptosis of some hepatocellular carcinoma cells. The reason of down-regulation of Vimentin protein may lie in that Caspase- Cracking. In this process, Jianpi Yiqi prescription can induce the apoptosis of hepatocarcinoma cell by using positive feedback and pro-apoptotic signal formed by Caspase-3 / Vimentin cleavage fragment and on the other hand may lead to the downregulation of NLRP3 / Caspase-1 inflammation by Vimentin level Weakening signal pathways, liver cancer intervention microenvironment.