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  

Mechanism of saikosaponin D in inhibiting the proliferation of anaplastic thyroid cancer cells via NLRP3-mediated pyroptosis

作　　者：高芳 李君 郭艳霞 李红 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.)

机构地区：[1]唐山市丰南区中医医院药剂科,河北唐山063300 [2]唐山市丰南区中医医院制剂室,河北唐山063300 [3]唐山市中医医院药学部,河北唐山063000

出　　处：《湖南中医药大学学报》2024年第10期1794-1800,共7页Journal of Hunan University of Chinese Medicine

基　　金：河北省中医药管理局科研计划项目(2021421)。

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、37μmol/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(0μmol/L柴胡皂苷D)、低浓度组(2.2μmol/L柴胡皂苷D)、中浓度组(11μmol/L柴胡皂苷D)、高浓度组(22μmol/L柴胡皂苷D)、si-NC组(转染NC siRNA)、高浓度+si-NC组(22μmol/L柴胡皂苷D联合转染NC siRNA)、高浓度+si-NLRP3组(22μmol/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。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(1,4,7,10,13,16,19,22,25,28,31,34,37μmol/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(0μmol/L saikosaponin D),low concentration group(2.2μmol/L saikosaponin D),medium concentration group(11μmol/L saikosaponin D),high concentration group(22μmol/L saikosaponin D)and si-NC group(transfected with NC)siRNA),high concentration+si-NC group(22μmol/L saikosaponin D co-transfected with NC siRNA),and high concentration+si-NLRP3 group(22μmol/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 NLR

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 

分 类 号：R285.5[医药卫生—中药学]