大黄酚通过激活NLRP3炎症小体诱导胃癌细胞焦亡的机制研究

目的探索大黄酚诱导人胃癌细胞焦亡的作用及其可能的作用机制。方法不同浓度大黄酚（0、5、10 μmol/L）和NLRP3抑制剂、caspase-1抑制剂先后处理人胃癌MKN28细胞，CCK-8法、克隆形成实验、流式细胞术检测细胞焦亡；实时荧光定量PCR检测焦亡、NLRP3炎症小体相关蛋白的mRNA水平；Western blotting检测焦亡、NLRP3炎症小体的蛋白表达水平。结果不同浓度的大黄酚（0、5、10 μmol/L）作用于MKN28细胞后，均降低了细胞存活率（P < 0.01），并促进了细胞焦亡的发生（P < 0.01）。大黄酚呈剂量依赖性地上调胃癌MKN28细胞中NLRP3、caspase-1和白细胞介素-1β的mRNA以及蛋白表达水平（P < 0.01）。采用caspase-1抑制剂VX-765、NLPR3抑制剂MCC950抑制NLPR3炎症小体表达，可削弱大黄酚对细胞焦亡的诱导作用（P < 0.01）。结论大黄酚抑制人胃癌MKN28细胞增殖、促进LDH释放和caspase-1活性升高，诱导胃癌细胞焦亡，其作用机制与NLRP3/caspase-1通路密切相关。

Study on the mechanism of chrysophanol inducing pyroptosis of gastric cancer cells by activating NLRP3 inflammasome

ObjectiveTo explore the effects of chrysophanol on pyroptosis of human gastric cancer cells and its possible mechanism.MethodsThe human gastric cancer MKN28 cells were treated with chrysophanol at different concentrations(0, 5 and 10 μmol/L), NLRP3 inhibitor and caspase-1 inhibitor.The pyroptosis was detected using CCK-8 method, clone formation experiment and flow cytometry.The mRNA levels of pyroptosis and NLRP3 inflammasome-related proteins detected using real-time PCR.The protein expression levels of pyroptosis and NLRP3 inflammasome-related proteins were detected using Western blotting.ResultsThe different concentrations of chrysophanol(0, 5 and 10 μmol/L) could decrease the cell viability and promote the occurrence of pyroptosison of MKN28 cells(P < 0.01).In a dose-dependent manner, the chrysophanol increased the mRNA and protein expression levels of NLRP3, caspase-1 and IL-1β in MKN28 cells(P < 0.01).The caspase-1 inhibitor VX-765 and NLPR3 inhibitor MCC950 were used to inhibit the expression of NLPR3 inflammasome, which could reduce the induced effect of chrysophanol on pyrotosis(P < 0.01).ConclusionsThe chrysophanol can inhibit the proliferation, promote the release of LDH, increase the caspase-1 activity and induce the pyroptosis of gastric cancer MKN28 cells, and which might be related to the NLRP3/caspase-1 pathway.