Department of Obstetrics, Lianyungang Maternal and Child Health Hospital, Lianyungang, Jiangsu, China.
Department of Genetics and Prenatal Diagnosis, Lianyungang Maternal and Child Health Hospital, Lianyungang, Jiangsu, China.
Bioengineered. 2022 Apr;13(4):9455-9466. doi: 10.1080/21655979.2022.2060626.
The present study aimed to investigate the expression, role, and underlying mechanism of action of sirtuin 1 (SIRT1) in congenital hypothyroidism (CH). A CH model was established in rats, and neuronal cells were isolated from the hippocampal tissues of normal rats. Free thyroxine (fT4) and thyroid-Stimulating hormone (TSH) concentrations were determined to confirm CH model conduction. The cognitive behavior of rats with CH was examined using open field and forced swimming tests. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression levels of SIRT1, p53, B-cell lymphoma-extra-large (Bcl-xl), Bcl-2-associated X (Bax), and cytochrome c in the hippocampal tissues and neuronal cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and flow cytometry were performed to evaluate cell viability and apoptosis, respectively. The results revealed that SIRT1 was expressed at low levels in the hippocampal tissues of rats with CH. Moreover, overexpression of SIRT1 in the hippocampal tissues of rats with CH and improved rat behavior, while reducing the CH-induced nerve cell apoptosis. In addition, this overexpression increased the viability, inhibited apoptosis, and reduced the expression of p53, Bax, and cytochrome c, while increasing the expression of Bcl-xl in cultured neurons. In contrast, SIRT1-small interfering RNA exhibited the opposite effects in cultured neurons. In conclusion, SIRT1 plays a role in the occurrence and development of CH by regulating nerve cell apoptosis.
本研究旨在探讨沉默信息调节因子 1(SIRT1)在先天性甲状腺功能减退症(CH)中的表达、作用及其作用机制。建立了大鼠 CH 模型,并从正常大鼠海马组织中分离出神经元细胞。通过测定游离甲状腺素(fT4)和促甲状腺激素(TSH)浓度来确认 CH 模型的建立。采用旷场和强迫游泳试验检测 CH 大鼠的认知行为。采用逆转录-定量聚合酶链反应(RT-qPCR)和蛋白质印迹法检测海马组织和神经元细胞中 SIRT1、p53、B 细胞淋巴瘤-extra-large(Bcl-xl)、Bcl-2 相关 X(Bax)和细胞色素 c 的表达水平。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)比色法和流式细胞术分别评估细胞活力和细胞凋亡。结果显示,CH 大鼠海马组织中 SIRT1 表达水平降低。此外,CH 大鼠海马组织中 SIRT1 的过表达改善了大鼠行为,同时减少了 CH 诱导的神经细胞凋亡。此外,这种过表达增加了神经元的活力,抑制了凋亡,降低了 p53、Bax 和细胞色素 c 的表达,同时增加了培养神经元中 Bcl-xl 的表达。相反,SIRT1 小干扰 RNA 在培养神经元中表现出相反的作用。综上所述,SIRT1 通过调节神经细胞凋亡在 CH 的发生发展中发挥作用。