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我们以往的工作表明,脑内血管紧张素Ⅱ(AⅡ)作为一种抗阿片物质参与吗啡耐受和电针耐受。本工作探讨在此过程中脑内AⅡ的基因表达是否加速。采用酚抽提法提取大鼠脑组织总RNA,使用人工合成的寡脱氧核糖核酸探针进行打点杂交。放射自显影结果表明,多次皮下注射吗啡或连续数小时电针的大鼠脑血管紧张素原(Ang)mRNA含量明显升高。在IBAS图象分析仪上,测量各杂交点自显影曝光斑的积分光密度值(I.O.D.)表明,连续注射吗啡1d或4d使Ang mRNA分别增高1倍或8倍;电针3或6h使Ang mRNA分别增高5倍或13倍。说明大鼠经3—4h吗啡或电针处理,即可引起在转录水平上脑内Ang基因表达加强。
Our previous work showed that intracerebral angiotensin II (AII) is involved as an anti-opioid in morphine tolerance and electro-acupuncture tolerance. This work explores whether AⅡ gene expression in the brain is accelerated during this process. Phenol extraction method was used to extract total RNA from rat brain and dotted hybridization was performed using synthetic oligodeoxyribonucleic acid probe. The results of autoradiography showed that the contents of angiotensinogen (Ang) mRNA in rats after subcutaneous injection of morphine or electroacupuncture for several hours were significantly increased. The integral optical density (IOD) of self-developed exposure spots at each hybridization point was measured on an IBAS image analyzer. Angiotensinogen (Ang) mRNA was either 1-fold or 8-fold increased at 1 or 4 days after morphine injection. mRNA were increased 5 times or 13 times. It shows that after 3-4h morphine or electroacupuncture treatment, the expression of Ang gene in the brain can be increased at transcriptional level.