基于代谢组学探究高原低氧对甲状腺激素合成的影响  

To explore the effect of high-altitude hypoxia on thyroid hormone synthesis based on metabolomics

作　　者：戴重阳 林雪[2] 王雅轩 蒲小燕 Dai Chongyang;Lin Xue;Wang Yaxuan;Pu Xiaoyan(Medical Department of Qinghai University,Xining810016;West China Hospital of Sichuan University,Chengdu610044)

机构地区：[1]青海大学医学部,西宁810016 [2]四川大学华西医院,成都610044

出　　处：《安徽医科大学学报》2024年第11期1989-1997,共9页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:82160322);青海省科学技术厅应用基础研究基金项目(编号:2023-ZJ-746)。

摘　　要：目的通过类靶向代谢组学技术探究高原低氧对甲状腺激素(TH)合成的影响。方法20只SPF级雄性SD大鼠随机分为对照组(Control)和低氧组(Hypoxia)。通过模拟海拔6 km低压氧舱低氧胁迫48 h构建SD大鼠急性低氧损伤模型,检测大鼠体质量、动脉血氧分压(PaO_(2))、血氧饱和度(SaO_(2));利用类靶向代谢组学技术检测两组大鼠血清代谢产物后进行KEGG富集分析;RT-PCR和Western blot法检测TH合成通路中关键基因钠碘转运体(NIS)、甲状腺过氧化物酶(TPO)、甲状腺球蛋白(TG)表达水平;ELISA法检测大鼠动脉血清甲状腺素(T 4)含量及谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)表达水平;RT-PCR、Western blot及ELISA法检测炎症指标Toll样受体-4(TLR-4)、白细胞介素-6(IL-6)、磷酸化细胞核因子κB(NF-κB/p65)、肿瘤坏死因子-α(TNF-α)表达水平;RT-PCR和Western blot法检测大鼠甲状腺组织中促凋亡蛋白Bcl-2关联X蛋白(Bax)和凋亡抑制蛋白B细胞淋巴瘤/白血病-2(Bcl-2)表达水平。结果与Control组相比,Hypoxia组大鼠体质量、PaO_(2)、SaO_(2)明显降低(P<0.01);Hypoxia组大鼠动脉血清差异代谢物显著富集在TH合成通路中(P<0.01),通路终产物T 4含量明显下降(P<0.01),且大鼠甲状腺组织NIS、TPO、TG mRNA与蛋白表达水平均降低(P<0.05);ELISA验证结果显示T 4含量变化与上述结果完全一致。与Control组相比,Hypoxia组大鼠血清SOD、GSH-Px酶活性下降,MDA含量增加(P<0.01);TLR-4、IL-6、NF-κB/p65、TNF-αmRNA、蛋白表达水平及含量均升高(P<0.05),甲状腺组织中Bax mRNA与蛋白表达水平明显升高,Bcl-2则明显降低(P<0.05)。结论高原低氧环境暴露加剧机体氧化应激和炎症反应,促进甲状腺滤泡上皮细胞凋亡,从而影响甲状腺功能,最终导致T 4合成量降低。Objective To explore the effects of high-altitude hypoxia on thyroid hormone(TH)synthesis by quasi-targeted metabolomics technology.Methods Twenty SPF male SD rats were randomly divided into Control group and Hypoxia group.An acute hypoxia injury model was established in SD rats by simulating hypoxia stress in a hypobaric oxygen chamber at an altitude of 6 km for 48 hours.The body weight,arterial oxygen partial pressure(PaO_(2))and blood oxygen saturation(SaO_(2))were detected.KEGG enrichment analysis was performed after the metabolites in the blood of two groups were detected by quasi-targeted metabolomics technology.The expression levels of sodium iodide symporter(NIS),thyroid peroxidase(TPO)and thyroglobulin(TG)were detected by RT-PCR and Western blot in TH synthesis pathway.The content of serum thyroxine(T 4)and the expression levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by ELISA.The expression levels of toll-like receptors-4(TLR-4),interleukin-6(IL-6),nuclear factor-κB/p65(NF-κB/p65)and tumor necrosis factor-α(TNF-α)were detected by RT-PCR,Western blot and ELISA.The expression levels of Pro apoptotic protein Bcl-2 associated X protein(Bax)and inhibitor of apoptosis protein B-cell lymphoma/leukemia-2(Bcl-2)were detected by RT-PCR and Western blot in rats thyroid tissue.Results Compared with the Control group,the body weight,PaO_(2)and SaO_(2)of rats in the Hypoxia group significantly decreased(P<0.01).The differential metabolites in arterial serum of hypoxia group rats were significantly enriched in the TH synthesis pathway,and the content of the pathway end product T 4 decreased significantly(P<0.01).In addition,the mRNA and protein expression levels of NIS,TPO,TG in rats thyroid tissue significantly decreased(P<0.05).The ELISA validation results showed that the changes of T 4 content were completely consistent with the above results.Compared with the Control group,the enzyme activities of SOD and GSH-Px in the serum of rats in the hypoxia group

关 键 词：低氧 类靶向代谢组学 氧化应激 炎症 甲状腺激素 

分 类 号：R335.2[医药卫生—人体生理学] R339.5.4[医药卫生—基础医学] R339.6