文献类型：期刊文献

中文题名：山柰酚通过激活NLRP3-Caspase-1信号轴介导焦亡增强胃癌细胞对奥沙利铂的化疗敏感性

英文题名：Kaempferol Enhances Chemosensitivity of Gastric Cancer Cells to Oxaliplatin by Activating the NLRP3-Caspase-1 Signaling Axis to Mediate Pyroptosis

作者：陈凤琴[1];杨春婷[2,3];高夏青[2,3];黄存存[2,3];苏嵘[2,3];贾永男[1];陈怀霞[1];李海龙[1,2,3]

第一作者：陈凤琴

机构：[1]甘肃中医药大学附属医院,甘肃兰州730000;[2]甘肃中医药大学第一临床医学院,甘肃兰州730000;[3]甘肃中医方药挖掘与创新转化重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2026

卷号：37

期号：3

起止页码：447

中文期刊名：中药新药与临床药理

外文期刊名：Traditional Chinese Drug Research and Clinical Pharmacology

收录：;北大核心:【北大核心2023】；

基金：甘肃省青年科技基金计划项目(22JR5RA614);甘肃省卫生健康行业科研计划项目(GSWSKY2023-76)。

语种：中文

中文关键词：山柰酚;胃癌细胞;焦亡;奥沙利铂;化疗敏感性;NLRP3-Caspase-1信号轴;MKN-45细胞;HGC-27细胞

外文关键词：kaempferol;gastric cancer cells;pyroptosis;oxaliplatin;chemosensitivity;NLRP3-Caspase-1 signaling axis;MKN-45 cells;HGC-27 cells

摘要：目的探讨山柰酚通过调控NOD样受体热蛋白结构域相关蛋白3(NLRP3)-半胱氨酸天冬氨酸蛋白水解酶1(Caspase-1)信号轴促进胃癌细胞焦亡,从而增强胃癌细胞对奥沙利铂的化疗敏感性。方法CCK-8法检测山柰酚与奥沙利铂对胃癌细胞(MKN-45、HGC-27)增殖的影响。通过Synergy Finder在线软件检测山柰酚与奥沙利铂的协同效应。运用LDH释放实验检测LDH的活性。利用ELISA法检测白细胞介素1β(IL-1β)含量。借助扫描电镜和透射电镜观察细胞形态和细胞内超微结构。Western Blot技术检测NLRP3、Caspase-1、cleaved-Caspase-1、gasdermin D(GSDMD)、GSDMD-NT、gasdermin E(GSDME)、IL-18、IL-1β蛋白的表达。结果与对照组(0μmol·L^(-1))比较,山柰酚在20~140μmol·L^(-1)浓度范围、奥沙利铂在5~35μmol·L^(-1)浓度范围,分别干预24、48、72 h后,MKN-45、HGC-27细胞相对活力均明显降低(P<0.05,P<0.01),且呈现明显的时间、浓度依赖性。山柰酚与奥沙利铂联合干预HGC-27细胞的响应面模型以及相互作用效力(ZIP)指数为15.694,联合干预MKN-45细胞的ZIP指数为26.865,ZIP指数均>10,呈现协同效应。与对照组比较,山柰酚组的LDH和IL-1β释放量增加(P<0.05,P<0.01),且随着山柰酚浓度的增加,LDH和IL-1β释放量也增加(P<0.05,P<0.01)。与山柰酚组和奥沙利铂组比较,山柰酚组+奥沙利铂组LDH和IL-1β释放量增加(P<0.05,P<0.01)。用山柰酚和奥沙利铂干预后的两株胃癌细胞在扫描电镜和透射电镜下形态学特征不同,MKN-45细胞趋向于细胞焦亡形态学特征,而HGC-27细胞更趋向于铁死亡形态学特征。与对照组比较,不同浓度山柰酚干预两株胃癌细胞后,NLRP3、Caspase-1、cleaved-Caspase-1、GSDMD、GSDMD-NT、GSDME、IL-1β、IL-18蛋白表达水平均明显增高(P<0.05,P<0.01),且cleaved-Caspase1和GSDMD-NT表达量相对于Caspase-1和GSDMD增高较为明显。与空白对照组比较,山柰酚组、奥沙利铂组和山柰酚+奥沙利铂组NLRP3、Caspase-1、cleaved-Caspase-1、GSDMD、GSDMD-NT、GSDME、IL-1β、IL-18蛋白表达水平均明显增高(P<0.05,P<0.01),且cleaved-Caspase-1和GSDMD-NT表达量相对于Caspase-1和GSDMD增高较为明显,其中以山柰酚+奥沙利铂组效果最优(P<0.05,P<0.01)。结论山柰酚可能通过激活NLRP3-Caspase-1信号轴,促进胃癌细胞焦亡,从而增强胃癌细胞对奥沙利铂的敏感性。
Objective To investigate kaempferol enhances the chemosensitivity of gastric cancer cells to oxaliplatin by promoting pyroptosis through regulation of the NOD-like receptor thermal protein domain associated protein 3(NLRP3)-Caspase-1 signaling axis.Methods The effect of kaempferol and oxaliplatin on the proliferation of gastric cancer cells(MKN-45,HGC-27)was detected by CCK-8 assay.The synergistic effect between kaempferol and oxaliplatin was analyzed using the Synergy Finder online software.LDH release assay was used to measure LDH activity.Interleukin-1β(IL-1β)content was detected by ELISA.Cell morphology and intracellular ultrastructure were observed using scanning electron microscopy(SEM)and transmission electron microscopy(TEM).The protein expression levels of NLRP3,Caspase-1,cleaved-Caspase-1,gasdermin D(GSDMD),GSDMD-NT,gasdermin E(GSDME),IL-18,and IL-1βwere detected by Western Blot.Results Compared with the control group(0μmol·L^(-1)),after treatment with kaempferol(20-140μmol·L^(-1))or oxaliplatin(5-35μmol·L^(-1))for 24,48,and 72 hours,the relative viability of both cell lines was significantly reduced(P<0.05,P<0.01)in a time-and concentration-dependent manner.The ZIP synergy score for the combined treatment of HGC-27 cells with kaempferol and oxaliplatin was 15.694,and for MKN-45 cells was 26.865.Both scores were greater than 10,indicating a synergistic effect.Compared with the control group,the kaempferol-treated groups showed increased release of LDH and IL-1β(P<0.05,P<0.01),and the release increased with higher kaempferol concentrations(P<0.05,P<0.01).Compared with the kaempferol-only or oxaliplatin-only groups,the combined treatment group(kaempferol+oxaliplatin)showed increased release of LDH and IL-1β(P<0.05,P<0.01).The two gastric cancer cell lines treated with kaempferol and oxaliplatin exhibited different morphological features under SEM and TEM.MKN-45 cells tended to show morphological characteristics of pyroptosis,while HGC-27 cells leaned more towards features of ferroptosis.Compared with the control group,treatment of both gastric cancer cell lines with different concentrations of kaempferol significantly increased the protein expression levels of NLRP3,Caspase-1,cleaved-Caspase-1,GSDMD,GSDMD-NT,GSDME,IL-1β,and IL-18(P<0.05,P<0.01).The increase in the expression of cleaved-Caspase-1 and GSDMD-NT relative to their fulllength counterparts(Caspase-1 and GSDMD)was particularly notable.Compared with the blank control group,the kaempferol group,oxaliplatin group,and the combined treatment group all showed significantly increased protein expression levels of NLRP3,Caspase-1,cleaved-Caspase-1,GSDMD,GSDMD-NT,GSDME,IL-1β,and IL-18(P<0.05,P<0.01).Again,the increase in cleaved-Caspase-1 and GSDMD-NT was more pronounced relative to their pro-forms.The combined treatment group(kaempferol+oxaliplatin)showed the most potent effect(P<0.05,P<0.01).Conclusion Kaempferol may enhance the sensitivity of gastric cancer cells to oxaliplatin by promoting pyroptosis through activation of the NLRP3-Caspase-1 signaling axis.