摘要
目的:基于"病证结合"理论,构建和评价广泛性焦虑(generalized anxiety disorder,GAD)心阴亏虚证大鼠模型。方法:按随机数字表法将32只SD大鼠随机分为4组,每组8只,即空白组、模型组、天王补心丸组、地西泮片组。根据先疾病后证型的思路进行造模,除空白组外,其余3组大鼠通过不确定性空瓶应激法先建立GAD模型,再采用水环境小站台睡眠剥夺法形成大鼠心阴亏虚证候模型,以此构建GAD心阴亏虚证模型。造模成功后,空白组和模型组不给与任何处理,天王补心丸组、地西泮片组分别给予天王补心丸、地西泮片灌胃,1次/d,连续干预21 d。从宏观表征、微观指标、以方验证三个方面对模型进行评价。每日观察大鼠一般情况。造模前、造模后及干预第1、2、3周末采用高架十字迷宫实验、旷场实验观察大鼠行为学变化。造模前、造模后及干预第3周末检测大鼠收缩压、心率、呼吸、肛温水平。干预第3周末采用实时荧光定量聚合酶链式反应法(real-time PCR)检测大鼠海马GR mRNA表达水平。结果:(1)宏观表征:①行为学结果显示:与空白组比较,造模后、造模后1周、造模后2周、造模后3周模型组OE%值、OT%值、水平运动、垂直运动均降低(P<0.05)。②生命体征结果显示:与空白组比较,造模后、造模后3周模型组收缩压均较低(P<0.01);心率、呼吸、肛温均较高(P<0.01)。(2)微观指标:RT-PCR结果显示:造模后3周,与空白组比较,模型组大鼠海马GR mRNA表达水平降低(P<0.01)。(3)以方验证:①行为学结果:与模型组比较,天王补心丸组、地西泮片组干预后各时间点OE%值、OT%值、水平运动、垂直运动均升高(P<0.05);地西泮片组干预后各时间点OE%值、干预第1、2周末OT%值、水平运动和干预第1、2周末垂直运动均升高(P<0.01)。②生命体征结果:干预第3周末,与模型组比较,天王补心丸组、地西泮片组收缩压均升高(P<0.05),心率、呼吸、肛温均降低(P<0.05)。③RT-PCR结果:干预第3周末,与模型组比较,天王补心丸组、地西泮片组大鼠海马GR mRNA表达水平均升高(P<0.05)。结论:采用不确定性空瓶应激法结合水环境小站台睡眠剥夺法成功构建了GAD心阴亏虚病证结合模型,该模型较为稳定,可控性相对较强,值得进一步应用。
Objective:Based on the theory of "Combination of Disease with Syndrome", we established and evaluated the model of generalized anxiety disorder(GAD)of heart Yin deficiency in rats. Methods:According to the method of random number table, totally 32 male rats were randomly divided into4 groups with 8 rats in each group, including normal group, model group, Tianwang Buxin Pills group, Diazepam Tablets group. In addition to the normal group, the other groups' rats by the method of uncertainty empty bottle stimulation replicated the GAD model, and then we used the water environment with small platform by sleep deprivation method to create the syndrome of heart Yin deficiency in ratsso as to establish the model of heart Yin deficiency of GAD rats. After the model was successfully made, the normal group and the model group were not given any treatment, but the intervention group were given Tianwang Buxin Pills and diazepam tablets once a day for 21 days, respectively. The model was evaluated from three aspects: macroscopic representation, micro index and prescription validation model. The general condition of rats was recorded by daily observation. The ethology results of rats were observed before and after modeling and after 1,2,3 weeks of intervention with open field test and elevated plus-maze test. The systolic blood pressure, heart rate, breathing, anal temperatureresults of rats were observed before and after modeling and after 3 weeks of intervention. The expression of GR mRNA in hippocampus of rats was detected by real-time quantitative polymerase chain reaction(real-time PCR) after 3 weeks of intervention. Results:From macroscopic representation,the ethology results showed that compared with the normal group, the model group in the OE% value, the OT% value, the horizontal movement and the vertical movement all decreased significantly(P<0.05) after model was made and 1,2,3 weeks after modeling. The results of vital signs showed that systolic blood pressure in model group were lower than those in normal group(P<0.01).The heart rate, respiration and anus temperature were higher than those in normal group(P<0.01) after model was made and 3 weeks after modeling. From micro index,after 3 weeks modeling,the results of real-time PCR showed thatthe expression of GR mRNA in hippocampus of model group was significantly lower than that of normal group(P<0.01). From prescription validation model,the first was ethology results. Compared with the model group, the OE% value, the OT% value, the horizontal movement and vertical movement increased in Tianwang Buxin Pills group significantly at each time point after intervention(P<0.05). Compared with the model group, the Diazepam Tablets group had significant differences in the value of OE%, the horizontal movement at each time point, the OT% value and vertical movement at the first and second week(P<0.05).The second was the result of vital signs. Compared with the model group, the blood pressure was significantly increased(P<0.05), and the heart rate, respiration and anus temperature were significantly decreased in the Tianwang Buxin Pills group, Diazepam Tablets group(P<0.01). At last was the results of real-time PCR.The expression level of GR mRNA in hippocampus of Tianwang Buxin Pills group and Diazepam Tablets group was higher than that of model group(P<0.05). Conclusion:The model of GAD rats with heart Yin deficiency was established by using the method of uncertain empty bottle stress and small platform sleep deprivation in water environment. The model was relatively stable and controllable, which was worthy of further study.
引文
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