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为研究曲马多(tramadol,INN)作用下大鼠不同脑区AMPK(腺苷单磷酸活化蛋白激酶)表达量的变化,探讨INN的中枢镇痛机制,将30只SD纯种大鼠随机分为对照组和INN组(Q组),Q组又分为4个亚组:Q1组(注射INN后10min)、Q2组(注射INN后20min)、Q3组(注射INN后40min)和Q4组(注射INN后60min),各组大鼠到达预定的时间点后分别采取脑组织,应用RT-PCR法检测脑内AMPKα1、α2mRNA转录量,应用Western blot方法检测p-AMPK蛋白的相对表达量。结果显示:腹腔注射INN后引起大鼠各脑区AMPKα1、α2mRNA高效表达,各时期AMPKα1、α2mRNA表达与对照组比较差异显著(P<0.01或P<0.05);在小脑区p-AMPK蛋白的相对表达量在10min时间点与对照组比较差异极显著(P<0.01),在大脑皮层、丘脑和脑干区其相对表达量在40,60 min时间点与对照组比较差异显著(P<0.01或P<0.05),海马区则只在40min时间点差异极显著(P<0.01)。结果提示,大鼠各脑区的AMPK参与了INN的镇痛过程,而INN的镇痛机制可能与AMPK的高效表达有关。
In order to study the changes of AMPK (adenosine monophosphate activated protein kinase) in different brain regions of rats under the action of tramadol (INN), the central analgesic mechanism of INN was studied. Thirty SD SD rats were randomly divided into three groups (Control group) and INN group (Q group). Q group was divided into four subgroups: Q1 group (10 min after INN injection), Q2 group (20 min after INN injection), Q3 group (40 min after INN injection) and Q4 group (60min after injection of INN). Brain tissues were taken from each group at the predetermined time points. The transcription level of AMPKα1 and α2 mRNA in the brain was detected by RT-PCR. The relative expression of p-AMPK protein was detected by Western blot. The results showed that intraperitoneal injection of INN resulted in the high expression of AMPKα1 and α2 mRNA in rat brain. The expressions of AMPKα1 and α2 mRNA in each group were significantly different from those in control group (P <0.01 or P <0.05) Compared with the control group, the relative expression level in the cerebral cortex, thalamus and brainstem area at 40 min and 60 min was significantly different from that in the control group (P <0.01 or P <0.01) P <0.05), while the difference in the hippocampal area was only significant at 40min (P <0.01). The results suggest that AMPK in various brain regions of rats is involved in the analgesic process of INN, and the analgesic mechanism of INN may be related to the high expression of AMPK.