左旋四氢巴马汀和地西泮对条件性恐惧大鼠镇静及抗焦虑作用的比较

Comparison of the sedative and anti-anxiety effects of levo-tetrahydropalmatine and diazepam on rats with conditioned fear

目的比较左旋四氢巴马汀(levo-tetrahydropalmatine,L-THP)和地西泮在干预条件性恐惧模型大鼠中的镇静及抗焦虑作用。方法将32只雄性成年大鼠按照随机数字表法分为空白组、模型组、地西泮组和左旋四氢巴马汀组(L-THP组)(每组8只)。运用足底电击法复制条件性恐惧大鼠模型,造模4 d后,地西泮组和L-THP组分别给予地西泮(3.6 mg/kg)和L-THP(25 mg/kg)灌胃,1次/d,连续10 d,空白组和模型组给予等同体积的生理盐水。给药结束后运用高架十字迷宫实验和旷场实验测定大鼠的焦虑行为,运用睡眠能量监测系统检测睡眠和能量相关指标的变化。采用SPSS 23.0及Graphpad Prism 7.0进行数据分析,多组间比较采用单因素方差分析,两两比较用LSD检验。结果高架十字迷宫实验结果显示,与模型组比较,地西泮组大鼠开放臂进入次数百分比[(11.27±8.78)%,(30.11±14.59)%;P<0.05]和开放臂停留时间百分比[(1.94±1.48)%,(17.53±8.21)%;P<0.05]均显著高。与模型组比较,L-THP组大鼠开放臂停留时间、开放臂进入次数百分比和开放臂停留时间百分比较高,但差异无统计学意义(均P>0.05)。旷场实验结果显示,与模型组比较,地西泮组大鼠进入中央格时间[(2.99±1.83)s,(6.94±3.52)s;P<0.05]和进入周边场时间[(297.01±1.83)s,(293.30±3.52)s;P<0.05]均高;与模型组比较,L-THP组大鼠各指标变化均差异无统计学意义(均P>0.05)。自主活动实验结果显示,模型组大鼠夜晚自主活动次数比空白组少[(758.79±375.37)次/h,(1101.93±525.96)次/h;P<0.05]。与模型组比较,L-THP组大鼠白天自主活动次数[(502.40±228.54)次/h,(820.57±364.60)次/h;P<0.05]少、夜晚自主活动次数[(1146.85±309.69)次/h,(758.79±375.37)次/h;P<0.05]多,而全天自主活动次数未见显著性改变。各组大鼠能量代谢的相关指标与睡眠时间相关分析结果显示,大鼠白天睡眠时间与热量消耗值(heat值)呈负相关(r=-0.335,P<0.05),夜晚睡眠时间与白天heat值呈正相关(r=0.352,P<0.05)。结论左旋四氢巴马汀在本研究所用的浓度范围内未见显著的抗焦虑作用,但其镇静及改善睡眠活动节律作用优于地西泮。

Objective To compare the sedative and anti-anxiety effects of levo-tetrahydropalmatine(L-THP)and diazepam on conditioned fear model rats.Methods According to the random number table method,32 adult male rats were divided into blank group,model group,diazepam group and L-THP group(with 8 rats in each group).The conditioned fear model was reproduced by the plantar electric shock method.Four days after the modeling,the rats in diazepam group and L-THP group were given diazepam(3.6 mg/kg)and L-THP(25 mg/kg)were respectively gavaged once a day for 10 days,the rats in blank group and model group were given the same volume of saline.After the administration,the elevated plus maze test and the open field test were used to measure the anxiety behavior of the rats,and the sleep energy monitoring system was used to detect changes in sleep and energy-related indicators.SPSS 23.0 and Graphpad Prism 7.0 softwares were used for data analysis,multiple samples between groups were compared by one-way ANOVA,and LSD test was used for pairwise comparison.Results The results of the elevated plus maze experiment showed that compared with the model group,the percentage of open-arm entry times((11.27±8.78)%,(30.11±14.59)%,P<0.05)and the percentage of open-arm residence time((1.94±1.48)%,(17.53±8.21)%,P<0.05)in diazepam group were all significantly increased.Compared with the model group,the open arm residence time,the percentage of open arm residence time and the percentage of open arm entry times in L-THP group showed an upward trend,but there was no statistical significance(all P>0.05).The results of the open field experiment showed that compared with the model group,the time of entering the central grid((2.99±1.83)s,(6.94±3.52)s,P<0.05)and the time of entering the peripheral field((297.01±1.83)s,(293.30±3.52)s,P<0.05)in diazepam group both increased.Compared with the model group,there was no significant difference in the changes of various indexes in L-THP group(all P>0.05).The results of locomotor activities showed that the autonomic activity times of model group in nighttime was significantly lower than that of blank group((758.79±375.37)times/h,(1101.93±525.96)times/h,P<0.05).Compared with the model group,the number of autonomous activities of rats in L-THP group in daytime((820.57±364.60)times/min,(502.40±228.54)times/min,P<0.05)decreased,and the number of autonomous activities in the nighttime((758.79±375.37)times/min,(1146.85±309.69)times/min,P<0.05)increased,but there was no significant change in the number of autonomous activities in the whole day.Correlation analysis of energy metabolism related indexes and sleep time of rats in each group were analyzed.The experimental results showed that the daytime sleep time were negatively correlated with heat value(r=-0.335,P<0.05),and the night sleep time was positively correlated with daytime heat value(r=0.352,P<0.05).Conclusion L-tetrahydropalmatine has no significant anti-anxiety effect in the concentration range used in this study,but its sedative and improving sleep activity rhythm are better than diazepam.