Li Jia-Ping, Zhou Jing-Xuan, Wang Qi, Gu Gao-Qin, Yang Shi-Jin, Li Cheng-Ye, Qiu Chang-Wei, Deng Gan-Zhen, Guo Meng-Yao
Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
Biol Trace Elem Res. 2016 Sep;173(1):116-25. doi: 10.1007/s12011-016-0620-8. Epub 2016 Jan 18.
Selenium (Se), a nutritionally essential trace element, is associated with health and disease. Selenoprotein T (SelT) was identified as a redoxin protein with a selenocystein, localizing in the endoplasmic reticulum. The myosin light chain kinase (MLCK) and myosin light chain (MLC) play key roles in the contraction process of smooth muscle. The present study was to detect the effect and mechanism of SelT on the contraction process of gastric smooth muscle. The WT rats were fed with different Se concentration diets, and Se and Ca(2+) concentrations were detected in the gastric smooth muscle. Western blot and qPCR were performed to determine SelT, CaM, MLCK, and MLC expressions. MLCK activity was measured by identifying the rates of [γ-32P]ATP incorporated into the MLC. The results showed Se and Ca(2+) concentrations were enhanced with Se intake in gastric smooth muscle tissues. With increasing Se, SelT, CaM, MLCK and MLC expressions increased, and MLCK and MLC activation improved in gastric smooth muscle tissue. The SelT RNA interference experiments showed that Ca(2+) release, MLCK activation, and MLC phosphorylation were regulated by SelT. Se affected the gastric smooth muscle constriction by regulating Ca(2+) release, MLCK activation, and MLC phosphorylation through SelT. Se plays a major role in regulating the contraction processes of gastric smooth muscle with the SelT.
硒(Se)是一种营养必需的微量元素,与健康和疾病相关。硒蛋白T(SelT)被鉴定为一种含硒代半胱氨酸的氧化还原蛋白,定位于内质网。肌球蛋白轻链激酶(MLCK)和肌球蛋白轻链(MLC)在平滑肌收缩过程中起关键作用。本研究旨在检测SelT对胃平滑肌收缩过程的影响及其机制。给野生型大鼠喂食不同硒浓度的日粮,检测胃平滑肌中的硒和钙离子(Ca²⁺)浓度。采用蛋白质免疫印迹法(Western blot)和实时定量聚合酶链反应(qPCR)检测SelT、钙调蛋白(CaM)、MLCK和MLC的表达。通过测定[γ-³²P]ATP掺入MLC的速率来检测MLCK活性。结果显示,胃平滑肌组织中硒和Ca²⁺浓度随硒摄入量的增加而升高。随着硒含量的增加,胃平滑肌组织中SelT、CaM、MLCK和MLC的表达增加,MLCK和MLC的激活增强。SelT RNA干扰实验表明,Ca²⁺释放、MLCK激活和MLC磷酸化受SelT调控。硒通过SelT调节Ca²⁺释放、MLCK激活和MLC磷酸化,从而影响胃平滑肌收缩。硒通过SelT在调节胃平滑肌收缩过程中起主要作用。