摘要
目的探讨内质网应激和p38丝裂原活化蛋白激酶(P38MAPK)信号通路在高碘诱导甲状腺上皮细胞损伤中的作用。方法将甲状腺上皮细胞Nthy-ori 3-1随机分为对照组(正常培养)、模型组(40 mmol·L^(-1)碘化钾)、4-苯基丁酸(4-PBA)组[40 mmol·L^(-1)碘化钾和2 mmol·L^(-1)4-PBA]、SB203580组(40 mmol·L^(-1)碘化钾和10μmol·L^(-1)SB203580)。用蛋白质印迹法检测细胞葡萄糖调节蛋白78(GRP78)、p-P38/P38的表达水平;用MTT和克隆形成实验检测增殖水平;用流式细胞术检测细胞凋亡水平;用酶联免疫吸附试验法检测白细胞介素-6(IL-6)水平。结果对照组、模型组、4-PBA组和SB203580组GRP78蛋白表达水平分别为0.15±0.03、0.61±0.07、0.27±0.03和0.37±0.04;p-P38/P38比值分别为0.12±0.03、0.53±0.04、0.35±0.04和0.25±0.03;细胞存活率分别为(100.00±0.00)%、(53.71±6.16)%、(80.24±8.17)%和(71.29±7.36)%;克隆形成数分别为(271.36±25.18)、(96.09±10.79)、(183.24±15.36)和(141.24±16.18)个;凋亡率分别为(1.04±0.21)%、(9.27±1.67)%、(3.18±1.52)%和(3.82±1.09)%;IL-6水平分别为(0.71±0.08)、(9.17±0.87)、(3.26±0.29)和(4.71±0.41)nmol·L^(-1)。上述指标,模型组和对照组比较,在统计学上差异均有统计学意义(均P<0.05);4-PBA组、SB203580组和模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论高碘可抑制Nthy-ori 3-1细胞增殖,诱导细胞凋亡和炎症因子分泌,其机制可能与高碘激活内质网应激和P38MAPK信号通路有关。
Objective To investigate the role of endoplasmic reticulum stress and p38 mitogen-activated protein kinase P38MAPK signaling pathway in thyroid epithelial cell injury induced by high iodine.Methods The thyroid epithelial cells Nthy-ori 3-1 were randomly divided into control group(normal culture),model group(40 mmol·L^(-1)potassium iodide),4-phenylbutyric acid(4-PBA)group(40 mmol·L^(-1)potassium iodide and 2 mmol·L^(-1)4-PBA)and SB203580 group(40 mmol·L^(-1)potassium iodide and 10μmol·L^(-1)SB203580).Western blot was used to detect the expression of glucose regulated protein 78(GRP78)and p-P38/P38 of Nthy-ori 3-1 cells.MTT and colony formation experiments were used to detect the proliferation level.Flow cytometry was used to detect the apoptosis level.Enzymelinked immunosorbent assay(ELISA)was used to detect the level of interleukin-6(IL-6).Results The expression levels of GRP78 protein in control group,model group,4-PBA group and SB203580 group were0.15±0.03,0.61±0.07,0.27±0.03 and 0.37±0.04;the ratios of p-P38/P38 were 0.12±0.03,0.53±0.04,0.35±0.04 and 0.25±0.03;cell survival rates were(100.00±0.00)%,(53.71±6.16)%,(80.24±8.17)%and(71.29±7.36)%;the number of clones formed was 271.36±25.18,96.09±10.79,183.24±15.36 and141.24±16.18;the apoptosis rates were(1.04±0.21)%,(9.27±1.67)%,(3.18±1.52)%and(3.82±1.09)%;IL-6 secretion levels were(0.71±0.08),(9.17±0.87),(3.26±0.29)and(4.71±0.41)nmol·L^(-1),respectively.For the above indicators,there was significant difference between the model group and the control group(all P<0.05);there was significant difference between the 4-PBA group,SB203580 group and the model group(all P<0.05).Conclusion High iodine can inhibit the proliferation of Nthy-ori 3-1 cells and induce apoptosis and secretion of inflammatory factors,which may be related to the activation of endoplasmic reticulum stress and P38MAPK signaling pathway by high iodine.
作者
杨鑫娜
蔡辉耀
陈丽君
梁波
YANG Xin-na;CAI Hui-yao;CHEN Li-jun;LIANG Bo(Department of Endocrinology,The Second Attached Hospital of Fujian Medical University,Quanzhou 362000,Fujian Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2024年第16期2339-2343,共5页
The Chinese Journal of Clinical Pharmacology
基金
福建省中青年教师教育科研项目(科技类)立项基金资助项目(JAT200163)。
关键词
高碘
内质网应激
P38丝裂原活化蛋白激酶
甲状腺上皮细胞
凋亡
high iodine
endoplasmic reticulum stress
p38 Mitogen-Activated Protein Kinase
thyroid epithelial cell
apoptosis