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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
- 北京中医药 2024年43卷第12期 页码：1385-1391
- 作者机构：
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
- 作者简介：
李会敏，女，29岁，博士，住院医师。研究方向：中医药治疗糖尿病及甲状腺疾病。
魏军平，E-mail：weijunping@126.com
- 基金信息：国家自然科学基金资助项目(82074412);中央高水平中医院医院临床科研业务领军人才培育项目(CZ40905);中央高水平中医医院临床研究和成果转化能力提升项目(HLCMHPP2023084);2020年度首都卫生发展科研专项-自主创新(首发2020-2-4154)
DOI：10.16025/j.1674-1307.2024.12.011
中图分类号：纸质出版日期：2024-12-25，
收稿日期：2024-07-20，
- 稿件说明：
移动端阅览
李会敏，李宇思，任静，等.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［J］.北京中医药，2024，43（12）：1385-1391.
LI Huimin,LI Yusi,REN Jing,et al.Effects of serum containing Xiapu Xiaoying Formula on pyroptosis of human thyroid epithelial cells induced by high iodine[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(12):1385-1391.
李会敏，李宇思，任静，等.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［J］.北京中医药，2024，43（12）：1385-1391. DOI： 10.16025/j.1674-1307.2024.12.011.
LI Huimin,LI Yusi,REN Jing,et al.Effects of serum containing Xiapu Xiaoying Formula on pyroptosis of human thyroid epithelial cells induced by high iodine[J]. Beijing Journal of Traditional Chinese Medicine,2024,43(12):1385-1391. DOI： 10.16025/j.1674-1307.2024.12.011.
摘要
目的
2
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
2
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
2
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
2
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
2
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
2
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
2
Compared with the blank control group， cell viability in the model group was significantly reduced （
P
<
0.01）. Com
pared 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
2
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
Keywords
Xiapu Xiaoying Formulahigh iodinecell pyroptosisoxidative stress
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