夏朴消瘿方含药血清对高碘诱导的人甲状腺上皮细胞焦亡的影响
Effects of serum containing Xiapu Xiaoying Formula on pyroptosis of human thyroid epithelial cells induced by high iodine
李会敏 1李宇思 2任静 3肖瑶 1魏军平1
作者信息
- 1. 中国中医科学院广安门医院内分泌科,北京 100053
- 2. 西苑医院苏州医院内分泌科,苏州 215009
- 3. 国家电网公司北京电力医院儿科,北京 100073
- 折叠
摘要
目的 观察夏朴消瘿方(XPXYF)含药血清对碘化钠(NaI)诱导的人甲状腺上皮细胞(Nthy ori 3-1)焦亡的影响.方法 将Nthy ori 3-1细胞分为3组,空白组不予任何干预措施,模型组用NaI诱导细胞焦亡模型,实验组在造模的基础上加入含药血清培养.CCK-8法筛选含药血清最佳浓度,流式细胞术检测Nthy ori 3-1细胞焦亡,LDH法检测细胞毒性,细胞电镜观察Nthy ori 3-1细胞形态变化,流式细胞术检测氧化应激指标[活性氧(ROS)],Western blotting法检测细胞焦亡相关蛋白[焦亡蛋白D(GSDMD)、核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、Caspase-1、核因子κB(NF-κB)]表达.结果 与空白对照组比较,模型组细胞活力低(P<0.01);与模型组比较,实验组细胞活力高(P<0.01),实验组10%含药血清干预48 h,Nthy ori 3-1细胞活力最高.因此将10%含药血清用于后续研究.与空白对照组比较,模型组FLICA(+)-PI(+)双阳性比例高,焦亡率高(P<0.01).与模型组比较,实验组FLICA(+)-PI(+)双阳性比例低,焦亡率低(P<0.05).与空白对照组比较,模型组Nthy ori 3-1细胞LDH释放量多(P<0.01);与模型组比较,实验组细胞毒性少(P<0.01).实验组甲状腺滤泡细胞形态较规则,细胞核变小,胞质内空泡形成,空泡较模型组少,线粒体呈椭圆形,线粒体结构比模型组较完整.与空白对照组比较,模型组ROS水平高(P<0.01);与模型组比较,实验组ROS水平低(P<0.01).与空白对照组比较,模型组GSDMD、NF-κB p56蛋白表达高(P<0.05);与模型组比较,实验组GSDMD、Caspase-1、NF-κB p56、NLRP3蛋白表达低(P<0.05).结论 XPXYF含药血清可改善高碘诱导的Nthy ori 3-1细胞焦亡,可能与抑制氧化应激及激活GSDMD/NF-κB信号通路有关.
Abstract
Objective To observe the effect of serum containing Xiapu Xiaoying Formula(XPXYF)on sodium iodide(NaI)-induced pyroptosis in human thyroid epithelial cells(Nthy ori 3-1).Methods Nthy ori 3-1 cells were divided into three groups.The blank control group received no intervention,the model group was treated with NaI to establish a cell pyroptosis model,and the experimental group was cultured with drug-containing serum based on the established model.The optimal concentration of drug-containing serum was determined using the CCK-8 assay.Pyroptosis of Nthy ori 3-1 cells was detected using flow cytometry.Cytotoxicity was assessed,and morphological changes in Nthy ori 3-1 cells were observed under an electron microscope.Oxidative stress indicators[reactive oxygen species(ROS)]were measured by flow cytometry,and pyroptosis-related proteins[gasdermin D(GSDMD),nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),Caspase-1,and nuclear factor κB(NF-κB)]were analyzed using Western blotting.Results Compared with the blank control group,cell viability in the model group was significantly reduced(P<0.01).Compared with the model group,cell viability in the experimental group was significantly increased(P<0.01),with the highest viability observed after 48 hours of treatment with 10%drug-containing serum.Therefore,10%drug-containing serum was used for subsequent studies.Compared with the blank control group,the proportion of FLICA(+)-PI(+)double-positive cells and the pyroptosis rate were significantly higher in the model group(P<0.01).Compared with the model group,the experimental group showed a significantly lower proportion of FLICA(+)-PI(+)double-positive cells and pyroptosis rate(P<0.05).Compared with the blank control group,LDH release in the model group was significantly higher(P<0.01),while cytotoxicity in the experimental group was significantly lower than that in the model group(P<0.01).Morphological observation revealed that thyroid follicular cells in the experimental group were more regular in shape,with smaller nuclei,fewer vacuoles in the cytoplasm compared to the model group,oval mitochondria,and a more intact mitochondrial structure.Compared with the blank control group,ROS levels in the model group were significantly elevated(P<0.01),whereas ROS levels in the experimental group were significantly reduced compared to the model group(P<0.01).Compared with the blank control group,the expression levels of GSDMD and NF-κB p56 proteins were significantly increased in the model group(P<0.05).Compared with the model group,the experimental group showed significantly reduced expression levels of GSDMD,Caspase-1,NF-κB p56,and NLRP3 proteins(P<0.05).Conclusion XPXYF-containing serum can alleviate NaI-induced pyroptosis in Nthy ori 3-1 cells,potentially through inhibition of oxidative stress and suppression of the GSDMD/NF-κB signaling pathway activation.
关键词
夏朴消瘿方/高碘/细胞焦亡/氧化应激Key words
Xiapu Xiaoying Formula/high iodine/cell pyroptosis/oxidative stress引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据