逍遥丸改善产前应激子代大鼠焦虑样行为的作用机制

Mechanism of Xiaoyao Pill in Improving Prenatal Stress Induced Anxiety-like Behavior in Offspring Rats

目的探讨逍遥丸改善产前应激子代大鼠焦虑样行为的作用机制。方法采用随机数字表法选择28只SD子代大鼠,根据是否应激、灌胃不同药物,将大鼠分为对照组、产前应激组、产前应激+逍遥丸组和产前应激+氯化钠组,每组7只。然后通过旷场实验和高架十字实验观察各组子代大鼠的焦虑样行为,随后提取海马组织,分别采用荧光定量聚合酶链反应(PCR)和Western blot法观察谷氨酸受体A1亚基(GluA1)信使RNA(mRNA)和蛋白表达的变化。结果在旷场实验中,与对照组相比,产前应激组潜伏期明显延长,中央区时间百分比和运动距离明显减少(P<0.01);与产前应激+氯化钠组相比,产前应激+逍遥丸组子代大鼠潜伏期明显缩短,中央区时间百分比和运动距离明显增加[(55±14)s比(108±10)s、(14.8±2.4)%比(6.6±1.8)%、(2673±296)cm比(1747±353)cm](P<0.01);但各组子代大鼠运动速度比较差异无统计学意义(P>0.05)。在高架十字迷宫实验中,与对照组相比,产前应激组子代大鼠开放臂运动距离和开放臂时间百分比明显减少(P<0.01);与产前应激+氯化钠组相比,产前应激+逍遥丸组开放臂运动距离和开放臂时间百分比明显增加[(157±20)cm比(21±3)cm、(0.74±0.14)%比(0.50±0.15)%](P<0.01);但各组子代大鼠运动速度比较差异无统计学意义(P>0.05)。荧光定量PCR和Western blot法结果显示,与对照组相比,产前应激组子代大鼠海马中GluA1 mRNA和蛋白表达显著降低(P<0.01);与产前应激+氯化钠组相比,产前应激+逍遥丸组子代大鼠海马中GluA1 mRNA和蛋白表达显著升高(0.950±0.199 vs 0.491±0.181,0.663±0.047 vs 0.464±0.074)(P<0.01)。结论逍遥丸可明显改善产前应激子代大鼠焦虑样行为,其作用机制可能与提高海马区GluA1的表达有关。

Objective To explore the mechanism of Xiaoyao pill in improving anxiety-like behavior in offspring rats of prenatal stress.Methods A total of 28 SD offspring rats were selected,and were divided into a control group,a prenatal stress group,a prenatal stress+Xiaoyao pill group and a prenatal stress+sodium?chloride group according to different stress and gavage drugs,7 rats in each group.Anxiety-like behavior of each group of the offspring rats were observed through open field experiment and elevated cross experiments.Subsequently,hippocampal tissue was extracted and changes in glutamate receptor A1 subunit(GluA1)messenger RNA(mRNA)and protein expression were observed by fluorescence quantitative-polymerase chain reaction(RT-PCR)and Western blot methods respectively.Results In the open field experiment,compared with the control group,the incubation period of the prenatal stress group was significantly prolonged,and the percentage of time and movement distance in the central region were significantly reduced(P<0.01).Compared with the prenatal stress+sodium chloride group,the incubation period of the offspring rats of the prenatal stress+Xiaoyao pill group was significantly shortened,while the percentage of time and movement distance in the central region were significantly increased[(55±14)s vs(108±10)s,(14.8±2.4)%vs(6.6±1.8)%,(2673±296)cm vs(1747±353)cm](P<0.01).However,there was no significant difference in the movement speed of offspring rats among all groups(P>0.05).In the elevated cross maze experiment,compared with the control group,the open arm movement distance and open arm time percentage of the offspring rats in the prenatal stress group were significantly reduced(P<0.01).Compared with the prenatal stress+sodium chloride group,the percentage of open arm movement distance and open arm time in the prenatal stress+Xiaoyao pill group were significantly increased[(157±20)cm vs(21±3)cm,(0.74±0.14)%vs(0.50±0.15)%](P<0.01).However,there was no significant difference in the movement speed of the offspring rats among all groups(P>0.05).The results of fluorescence quantitative-PCR and Western blot showed that compared with the control group,GluA1 mRNA and protein expression in the hippocampus of offspring rats in the prenatal stress group were significantly decreased(P<0.01).Compared with the prenatal stress+sodium chloride group,GluA1 mRNA and protein expression in the hippocampus of offspring rats in the prenatal stress+Xiaoyao pill group were significantly increased(0.950±0.199 vs 0.491±0.181,0.663±0.047 vs 0.464±0.074)(P<0.01).Conclusion Xiaoyao pill can significantly improve anxiety-like behavior in prenatal stress offspring rats,and its mechanism of action may be related to increasing the expression of GluA1 in the hippocampus.