Hu Xiao-lin, Pan Ping-xi, Yang Li, Liu Xiao-jing, Fu Hua
Department of Internal Medicine, West China Hospital, Sichuan Univeristy, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2009 Mar;40(2):260-2, 267.
To investigate whether hypoxia could induce the expression of MIF in vascular smooth muscle cells (VSMCs).
Rat vascular smooth muscle cells (A7r5 from ATCC) were cultured under hypoxia condition (37 degrees C, 5% CO2, 1% O2) and normoxia condition (37 degrees C, 5% CO2, 20% O2) for 2, 4, 12, 24 or 48 hours respectively. MIF mRNA expression was detected by reverse transcriptase and polymerase chain reaction (RT-PCR). MIF protein expression was determined by Enzyme-Linked Immunosorbent Assay (ELISA).
Expression of MIF mRNA was up-regulated as early as 2 hours in VSMCs after being exposed to hypoxia condition, which remained the elevated level up to 48 hours. Meanwhile, MIF protein level increased after hypoxia treatment for 2 hours, and maintained elevated status up to 48 hours.
Hypoxia could significantly induce MIF expression in VSMCs. MIF might be an important factor in the patho-physiological response of VSMCs to hypoxia.
研究缺氧是否能诱导血管平滑肌细胞(VSMCs)中巨噬细胞移动抑制因子(MIF)的表达。
将大鼠血管平滑肌细胞(ATCC的A7r5)分别在缺氧条件(37℃,5%二氧化碳,1%氧气)和常氧条件(37℃,5%二氧化碳,20%氧气)下培养2、4、12、24或48小时。通过逆转录聚合酶链反应(RT-PCR)检测MIF mRNA表达。采用酶联免疫吸附测定(ELISA)法测定MIF蛋白表达。
VSMCs暴露于缺氧条件后,最早在2小时MIF mRNA表达上调,直至48小时仍维持在升高水平。同时,缺氧处理2小时后MIF蛋白水平升高,并持续升高至48小时。
缺氧可显著诱导VSMCs中MIF的表达。MIF可能是VSMCs对缺氧病理生理反应中的一个重要因素。