Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method.
Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method.
复制标题
使用半定量 RT-PCR 方法评估梗阻兔膀胱中的应激基因 mRNA（HSP-27、60 和 70）。
DOI：
10.1007/bf00929496
复制
发表时间：
1995
影响因子：
4.3
通讯作者：
Levin,RM
中科院分区：
文献类型：
--
作者：
Zhao,Y;Wein,AJ;Levin,RM
应激蛋白（Stress Proteins，HSPs）参与细胞对高温、缺氧和损伤等多种应激的反应。以前的工作使用北方印迹分析表明，增加表达的应激蛋白70（HSP-70）在兔膀胱组织受到部分出口梗阻。为了确定增加的表达是否是HSP-70特异性的，或者，替代地，指示广义的应激蛋白反应，使用改良的定量RT-PCR技术来定量正常和阻塞的兔膀胱组织中的HSP mRNA（HSP-27、60和70）。结果表明：1）改良的半定量RT-PCR技术是检测膀胱组织mRNA的一种灵敏、重复性好的方法。2)在正常膀胱组织中检测到HSP-27、HSP-60和HSP-70mRNA的组成性表达，其中HSP-70的相对信号强度最高，HSP-27的相对信号强度最低。HSP-27、60和70的mRNA在梗阻后24小时分别增加4.3、5.6和2.4倍，然后在梗阻后一周结束时逐渐恢复到对照水平，并在梗阻后14天内保持稳定。这些数据表明，改良的定量RT-PCR是一种有用的技术，用于检测膀胱组织中的mRNA;应力反应，发生在兔膀胱组织中的部分出口梗阻后是一种普遍现象。
Stress proteins (HSPs) participate in the cellular response to various stresses including hyperthermia, hypoxia and injury. A previous work using northern blot analysis demonstrated increased expression of stress protein 70 (HSP-70) in rabbit bladder tissue subjected to partial outlet obstruction. In order to determine if the increased expression was specific for HSP-70 or, alternatively, indicated a generalized stress protein response, a modified quantitative RT-PCR technique was used to quantitate HSP mRNAs (HSP-27, 60, and 70) in normal and obstructed rabbit urinary bladder tissues. The results show the following: 1) The modified semi-quantitative RT-PCR is a sensitive and reproducible technique for detecting mRNA in bladder tissue. 2) Constitutive levels of HSP-27, HSP-60, and HSP-70 mRNAs were detected in control bladder tissues; the relative signal intensity was highest for HSP-70 and lowest for HSP-27.3) A transient increase in HSP mRNAs was observed after obstruction; the mRNAs of HSP-27, 60 and 70 increased 4.3-, 5.6-, and 2.4-fold, respectively, at 24 h following obstruction, then gradually returned to control levels by the end of one week post-obstruction and remained stable up to 14 days post-obstruction. These data indicate that the modified quantitative RT-PCR is a useful technique for detecting mRNA in bladder tissue; the stress response which occurs in rabbit urinary bladder tissue following partial outlet obstruction is a general phenomenon.