Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041, PR China.
Auton Neurosci. 2013 Dec;179(1-2):84-9. doi: 10.1016/j.autneu.2013.08.066. Epub 2013 Aug 27.
The present experiments were carried out to investigate the expression of 3-mercaptopyruvate sulfurtransferase (3MST) in medulla oblongata of rats and effects of chronic intermittent hypoxia (CIH) on its expression. Sprague Dawley adult rats were randomly divided into two groups, including control (Con) group and CIH group. The endogenous production of hydrogen sulfide (H2S) in medulla oblongata tissue homogenates was measured using the methylene blue assay method, 3MST mRNA and protein expression were analyzed by RT-PCR and Western blotting, respectively, and the expression of 3MST in the neurons of respiratory-related nuclei in medulla oblongata of rats was investigated with immunohistochemical technique. CIH elevated the endogenous H2S production in rat medulla oblongata (P<0.01). The RT-PCR and Western blotting analyses showed that 3MST mRNA and protein were expressed in the medulla oblongata of rats and CIH promoted their expression (P<0.01). Immunohistochemical staining indicated that 3MST existed in the neurons of pre-Bötzinger complex (pre-BötC), hypoglossal nucleus (12N), ambiguous nucleus (Amb), facial nucleus (FN) and nucleus tractus solitarius (NTS) in the animals and the mean optical densities of 3MST-positive neurons in the pre-BötC, 12N and Amb, but not in FN and NTS, were significantly increased in CIH group (P<0.05). In conclusion, 3MST exists in the neurons of medullary respiratory nuclei and its expression can be up-regulated by CIH in adult rat, suggesting that 3MST-H2S pathway may be involved in regulation of respiration and protection on medullary respiratory centers from injury induced by CIH.
本实验旨在研究 3-巯基丙酮酸硫转移酶(3MST)在大鼠延髓中的表达及其在慢性间歇性低氧(CIH)中的作用。成年 Sprague Dawley 大鼠随机分为对照组(Con)和 CIH 组。采用亚甲基蓝法测定延髓组织匀浆中内源性硫化氢(H2S)的生成,采用 RT-PCR 和 Western blot 分析 3MST mRNA 和蛋白的表达,采用免疫组化技术观察大鼠延髓呼吸相关核团神经元中 3MST 的表达。CIH 可增加大鼠延髓内源性 H2S 的产生(P<0.01)。RT-PCR 和 Western blot 分析显示 3MST mRNA 和蛋白在大鼠延髓中表达,CIH 可促进其表达(P<0.01)。免疫组化染色显示 3MST 存在于大鼠前 Bötzinger 复合体(pre-BötC)、舌下神经核(12N)、疑核(Amb)、面核(FN)和孤束核(NTS)神经元中,CIH 组 pre-BötC、12N 和 Amb 神经元中 3MST 阳性神经元的平均光密度明显增加(P<0.05),而 FN 和 NTS 神经元中 3MST 阳性神经元的平均光密度无明显变化。综上所述,3MST 存在于延髓呼吸核团的神经元中,CIH 可上调其在成年大鼠中的表达,提示 3MST-H2S 通路可能参与 CIH 所致呼吸调节和延髓呼吸中枢损伤的保护。