摘要
目的 探究交泰丸自微乳对失眠大鼠下丘脑内5-羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopam-ine,DA)及去甲肾上腺素(noradrenaline,NE)等神经递质和肠道菌群的影响,探讨交泰丸自微乳对失眠的干预作用与作用机制.方法 采用对氯苯丙氨酸腹腔注射法建立失眠大鼠模型,灌胃给予模型大鼠交泰丸自微乳、黄连自微乳、肉桂自微乳,以地西泮、酸枣仁汤分别为阳性对照药,观察并记录各组动物行为学特征,并取下丘脑测定5-HT、NE、DA相关神经递质的活性,采用高通量测序法测定大鼠肠道菌群变化.结果 模型组大鼠自主活动时间、前肢离地次数均高于正常组(t=8.935 4、10.568 2,P=0.005 2、0.004 7),治疗后各组大鼠较模型组均有改善.模型组大鼠下丘脑内单胺神经递质5-HT水平显著降低,NE、DA水平显著升高(t=8.4729、15.497 3、21.885 7,P=0.006 8、0.005 7、0.007 2).与模型组大鼠相比,地西泮组、酸枣仁汤组、单方黄连组、交泰丸自微乳高剂量组及交泰丸自微乳中剂量组下丘脑内5-HT水平显著升高(t=40.547 7、28.673 2、15.635 4、17.498 0、36.498 6,P=0.005 4、0.003 9、0.006 3、0.003 8、0.004 1),NE、DA水平显著下降(t=12.462 2、8.498 6、24.558 6、18.876 0、7.445 8,P=0.004 9、0.006 7、0.005 4、0.007 5、0.024 1;t=24.483 7、12.8112、24.446 4、32.454 1、16.839 0,P=0.000 1、0.003 4、0.000 1、0.000 1、0.000 7),交泰丸自微乳低剂量组脑内5-HT、NE、DA水平相比模型组差异均无统计学意义(t=16.854 3、24.385 5、45.186 5,P=0.086 4、0.105 7、0.176 4).Alpha多样性分析结果表明,模型组大鼠的 Observed spe-cies平均指数、Chao1平均指数、Shannon平均指数均显著降低,与模型组相比,其余各给药组的菌群丰度与多样性均有回调.Beta多样性分析结果表明,与正常组相比,模型组肠道菌群的物种结构差异较大,治疗后各组大鼠肠道菌群的结构均发生改变,菌群丰度也较模型组有了提高.结论 交泰丸自微乳能提高失眠大鼠下丘脑内5-HT水平,降低NE、DA的水平,同时可提高失眠大鼠肠道菌群丰度,二者变化具有相关性,交泰丸可能通过影响失眠大鼠肠道菌群丰度、回调神经递质水平从而发挥治疗失眠的作用.
Abstract
Objective To observe the effects of Jiaotai Pill self-microemulsion(JPSM)on the intestinal flora and neuro-transmitters including 5-hydroxytryptamine(5-HT),dopamine(DA)and noradrenaline(NE)etc in the hypothalamus of in-somnia rats,and evaluate the intervention effect of JPSM on insomnia and the mechanism.Methods Dl-4-chlorophenylalanine was intraperitoneally injected to establish the insomnia rat models.JPSM,unilateral coptis self-mi-croemulsion and cinnamon self-microemulsion were given to the rat models by gavage,with diazepam and jujube seed de-coction as the positive control drugs respectively.The ethological characteristics of rats each group were observed.The activity of 5-HT,NE and DA in hypothalamus were measured.The changes of intestinal flora was determined using high-throughput sequencing.Results The time of autonomous activity and times of forelimb off the ground in the model group were higher than those in the normal control group(t=8.935 4,10.568 2;P=0.005 2,0.004 7).After treatment,the values in each group decreased compared with the model group.In the model group,the content of 5-HT in the hypothalamus signi-ficantly decreased,while those of NE and DA significantly increased(t=8.472 9,15.497 3,21.885 7;P=0.006 8,0.005 7,0.007 2).Compared with the model group,the contents of 5-HT in diazepam group,Suanzaoren Decoction group,unilater-al Coptis group,JPSM high-and medium-dose groups increased significantly(t=40.547 7,28.673 2,15.635 4,17.498 0,36.498 6;P=0.005 4,0.003 9,0.006 3,0.003 8,0.004 1),while those of NE and DA decreased significantly(t=12.462 2,8.498 6,24.558 6,18.876 0,7.445 8;P=0.004 9,0.006 7,0.005 4,0.007 5,0.024 1 and t=24.483 7,12.811 2,24.446 4,32.454 1,16.839 0;P=0.000 1,0.003 4,0.000 1,0.000 1,0.000 7);there was no significant difference in the contents of 5-HT,NE and DA in the low-dose JPSM group compared with model group(t=16.854 3,24.385 5,45.186 5;P=0.086 4,0.105 7,0.176 4).Alpha diversity analysis showed that the average indexes of Observed species,Chaol and Shannon in the model group significantly reduced.Compared with the model group,the abundance and diversity of bacterial flora in the other administration groups recovered.Beta diversity analysis showed that,compared with the normal control group,the species structure of intestinal flora in the model group was significantly different,and the structures of intestinal flora in the other administration groups changed.Conclusion JPSM can increase the contents of 5-HT and DA,and decrease that of NE in hypothalamus of rats with insomnia.The decrease of NE can increase the intestinal flora abundance of insomnia rats,and there is a correlation between the two changes.Jiaotai Pill may play a role in treating insomnia by affecting the intest-inal flora abundance of insomnia rats and regulating the levels of neurotransmitters.
基金项目
黑龙江省"双一流"特色学科(HLJTSXK-2022-03)
黑龙江省卫生健康委项目(2018-482)
佳木斯大学科技创新团队项目(cxtd202103)
NETL
NSTL
万方数据