土贝母通过炎性小体调控caspase-1诱导肝癌细胞焦亡抑制肝癌发生发展的机制研究

Pyroptosis is a new kind of programmed cell death which characterized by the activation of caspase-1 and the release of a large number of inflammatory cytokines. Previous studies have shown that levels of activity of inflammasomes and caspase-1 in HCC tissues were significantly lower than those of adjacent tissues, and serum contained extractive of Bolbostemma Paniculatun can significantly increase pyroptosis and upregulate the expression of caspase-1.However, the mechnism of pyroptosis in HCC induced by Bolbostemma Paniculatun is not clear. Based the results of papers and previous research, the hypothesis of the inhibition of inflammasome and caspase-1 can promote the development of HCC and Bolbostemma Paniculatun can induce pyroptosis to inhibit the occurrence and development of HCC through regulation activity of inflammasome and caspase-1 is proposed. The study of correlation between inflammasomes and the activity of caspase-1 and the degree of HCC will be carried based on samples of HCC patients. The molecular mechanism of HCC pyroptosis induced by serum contained Bolbostemma Paniculatun through regulation expression of various inflammasomes, caspase-1, IL-1 beta and IL-18 are investigated in vitro. The effect and molecular mechanism of pyroptosis of HCC treated by Bolbostemma Paniculatun is explored by tumor-bearing nude mice. The effective metabolic component in Bolbostemma Paniculatun which play a key role in pyroptosis is explored through metabonomics. The molecular mechanism of pyroptosis in hepatocellular carcinoma cells induced by Bolbostemma Paniculatun will be elucidated and provide a new theory for treatment of HCC by Chinese medicine Bolbostemma Paniculatun.

细胞焦亡是一种新发现的程序性细胞死亡，其特征是caspase-1被激活，并伴有大量炎症因子的释放。前期研究显示，肝癌组织中多种炎性小体及caspase-1活性显著低于癌旁组织；土贝母含药血清能促进肝癌细胞焦亡、上调caspase-1表达。但是，肝癌细胞发生焦亡的机制及土贝母对肝癌细胞焦亡的影响及机制尚不清楚。本课题以肝癌患者组织样本为材料，研究炎性小体和caspase-1活性与肝癌病理程度相关性；通过调控肝癌细胞各种炎性小体、caspase-1、IL-1β及IL-18的表达，探讨土贝母含药血清诱导肝癌细胞焦亡的分子机制；以肝癌荷瘤裸鼠模型为材料，研究土贝母对肝癌细胞焦亡的作用及分子机制；采用代谢组学方法探索土贝母含药血清中对肝癌细胞焦亡发挥关键作用的有效代谢成分。本研究将阐明肝癌细胞焦亡的分子机制及土贝母的代谢产物诱导肝癌细胞发生焦亡的分子机制，为肝癌的中药治疗提供新的理论依据。

项目的背景:肝癌组织中多种炎性小体及caspase-1活性显著低于癌旁组织；土贝母含药血清能促进肝癌细胞焦亡、上调caspase-1表达。但是，肝癌细胞发生焦亡的机制及土贝母对肝癌细胞焦亡的影响及机制尚不清楚。.研究内容：.1.肝癌患者组织样本中炎性小体和 caspase-1 活性与肝癌病理程度的临床相关性分析；2.土贝母含药血清对肝癌细胞焦亡的影响及机制研究；3.基于网络药理学研究土贝母治疗肝癌的药理机制；4.土贝母水提物对H22荷瘤小鼠的抑癌作用及抑制肝癌侵袭和转移的药效学研究；5.土贝母中含药血清对肝癌细胞焦亡的影响及机制研究；6.采用系统药理学研究肝癌的靶点及土贝母治疗肝癌的药效物质基础研究。.重要结果：.1.通过免疫组织化学和组织芯片检测，人肝癌组织中 Caspase-1 的表达水平显著低于癌旁组织。 2.土贝母水提物对肝癌细胞中 caspase-1 的表达水平无显著影响，能显著促进肝癌细胞 caspase-1 的活性及表达。3.Caspase-1 、IL-18、 NLRP3 在人肝癌细胞中表达水平显著低于肝正常细胞。4. 土贝母含药血清能诱导肝癌细胞7404发生细胞焦亡，并能调控肝癌细胞7404相关焦亡蛋白表达。5.利用TCMSP、SwissTarget Forecast和SINTCH对TBM有效成分的假定靶点进行了鉴定，筛选出9个活性分子。6.采用GO和KEGG分析，筛选出12个土贝母和肝癌的共有靶点。.科学意义：揭示了细胞焦亡在肝癌发生发展中的重要作用及分子机制，为探寻新的肝癌治疗靶点提供了可靠的依据。深入研究了土贝母含药血清对肝癌细胞焦亡的影响，采用基于网络药理学研究技术，探究了土贝母治疗肝癌的药效物质基础及候选分子的药理机制，并开展土贝母提取物贝母素乙诱导人肝癌细胞凋亡的分子机制研究，为临床上应用土贝母治疗肝癌提供了坚实的理论依据，也为开发新的药物制剂提供了依据。