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高芳, 李君, 郭艳霞, 李红.柴胡皂苷D通过NLRP3介导细胞焦亡抑制未分化甲状腺癌细胞增殖的机制研究[J].湖南中医药大学学报英文版,2024,44(10):1794-1800.[Click to copy
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| 柴胡皂苷D通过NLRP3介导细胞焦亡抑制未分化甲状腺癌细胞增殖的机制研究 |
| 高芳,李君,郭艳霞,李红 |
| (唐山市丰南区中医医院药剂科, 河北 唐山 063300;唐山市丰南区中医医院制剂室, 河北 唐山 063300;唐山市中医医院药学部, 河北 唐山 063000) |
| 摘要: |
| 目的 研究柴胡皂苷D通过NOD样受体蛋白3（NOD-like receptor protein 3，NLRP3）介导细胞焦亡，抑制未分化甲状腺癌细胞增殖的作用及机制。方法 培养未分化甲状腺癌细胞株HTh83和KMH2，不同浓度柴胡皂苷D（1、4、7、10、13、16、19、22、25、28、31、34、37 μmol/L）作用48 h后检测细胞存活率，计算半数抑制浓度（50% inhibition concentration，IC50）；细胞分为对照组（0 μmol/L柴胡皂苷D）、低浓度组（2.2 μmol/L柴胡皂苷D）、中浓度组（11 μmol/L柴胡皂苷D）、高浓度组（22 μmol/L柴胡皂苷D）、si-NC组（转染NC siRNA）、高浓度+si-NC组（22 μmol/L柴胡皂苷D联合转染NC siRNA）、高浓度+si-NLRP3组（22 μmol/L柴胡皂苷D联合转染NLRP3 siRNA），处理48 h后检测克隆形成数目，NLRP3、裂解型Caspase-1、GSDMD氨基末端片段（GSDMD N terminal fragment, GSDMD-N）的表达水平及培养基中白细胞介素-1β（interleukin-1β, IL-1β）、白细胞介素-18（interleukin-18, IL-18）的水平。结果 柴胡皂苷D以浓度依赖的方式降低HTh83细胞和KMH2细胞的存活率（P<0.05）；低浓度组、中浓度组、高浓度组的细胞克隆数目低于对照组（P<0.05），NLRP3、裂解型Caspase-1、GSDMD-N的表达水平及培养基中IL-1β、IL-18的水平高于对照组（P<0.05）；高浓度+si-NLRP3组细胞的存活率及克隆数目高于高浓度+si-NC组（P<0.05），NLRP3、裂解型Caspase-1、GSDMD-N的表达水平及培养基中IL-1β、IL-18的水平低于高浓度+si-NC组（P<0.05）。结论 柴胡皂苷D能显著抑制未分化甲状腺癌细胞增殖，其作用机制可能与激活NLRP3，介导细胞焦亡有关。 |
| 关键词: 未分化甲状腺癌 柴胡皂苷D 增殖 NOD样受体蛋白3 焦亡 |
| DOI：10.3969/j.issn.1674-070X.2024.10.011 |
| Received:May 12, 2024 |
| 基金项目:河北省中医药管理局科研计划项目(2021421)。 |
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| Mechanism of saikosaponin D in inhibiting the proliferation of anaplastic thyroid cancer cells via NLRP3-mediated pyroptosis |
| GAO Fang, LI Jun, GUO Yanxia, LI Hong |
| (Department of Pharmacy, Tangshan Fengnan District Hospital of Chinese Medicine, Tangshan, Hebei 063300, China;Department of Pharmacy Preparation, Tangshan Fengnan District Hospital of Chinese Medicine, Tangshan, Hebei 063300, China;Department of Pharmacy, Tangshan Hospital of Chinese Medicine, Tangshan, Hebei 063300, China) |
| Abstract: |
| Objective To study the effects and mechanism of saikosaponin D in inhibiting the proliferation of anaplastic thyroid cancer cells via NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis. Methods Anaplastic thyroid cancer cell lines HTh83 and KMH2 were cultured and treated with various concentrations of saikosaponin D (1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37 μmol/L) for 48 h. Cell survival rate was subsequently measured and half-maximal inhibitory concentration (IC50) was calculated. The cells were divided into control group (0 μmol/L saikosaponin D), low concentration group (2.2 μmol/L saikosaponin D), medium concentration group (11 μmol/L saikosaponin D), high concentration group (22 μmol/L saikosaponin D) and si-NC group (transfected with NC) siRNA), high concentration+si-NC group (22 μmol/L saikosaponin D co-transfected with NC siRNA), and high concentration+si-NLRP3 group (22 μmol/L saikosaponin D co-transfected with NLRP3 siRNA). After 48 h of treatment, the colony formation was quantified, the expression levels of NLRP3, cleaved caspase-1, and GSDMD N terminal fragment (GSDMD-N) as well as the levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the medium were measured. Results Saikosaponin D reduced the survival rate of HTh83 cells and KMH2 cells in a concentration-dependent manner. The number of cell clones in low-, medium-, and high-concentration groups was lower than that in the control group (P<0.05), while the expression levels of NLRP3, cleaved caspase-1, and GSDMD-N, and the levels of IL-1β and IL-18 in the medium were higher than those in the control group (P<0.05). The survival rate and clone number in high concentration +si-NC group were higher than those in high concentration+si-NC group, while the expression levels of NLRP3, cleaved caspase-1, and GSDMD-N, and the levels of IL-1β and IL-18 in the medium were lower than those in high concentration+si-NC group (P<0.05). Conclusion Saikosaponin D significantly inhibits the proliferation of anaplastic thyroid cancer cells, and its mechanism is probably related to the activation of NLRP3 and the mediation of pyroptosis. |
| Key words: anaplastic thyroid cancer saikosaponin D proliferation Nod-like receptor protein 3 pyroptosis |
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