ABSTRACT
ObjectiveTo explore the mechanism of Huanglian Ejiaotang in intervening in insomnia based on 5-hydroxytryptamine （5-HT） system and gut microbiota. MethodFifty-five SPF-grade SD rats were randomly divided into normal group， model group， low-， medium-， and high-dose Huanglian Ejiaotang groups （1.925， 3.85， and 7.7 g·kg-1）， and Estazolam group （0.1 mg·kg-1）. Except for those in the normal group， the rats in the other five groups were subjected to sleep deprivation on a narrow platform for 12 hours daily for 21 consecutive days. After 14 days of drug intervention， the sleep， exploratory behavior， and depressive-like behavior of the rats were assessed using the pentobarbital sodium sleep synergistic test， the open field test， and the sugar preference test， respectively. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of 5-HT， 5-hydroxyindoleacetic acid （5-HIAA）， tryptophan hydroxylase （TPH）， and monoamine oxidase-A （MAO-A）. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression of the 5-HT1A receptor （5-HT1AR） and 5-HT2A receptor （5-HT2AR）. Differences in gut microbiota among the groups were assessed using 16S rRNA sequencing， and the correlation between the 5-HT system and microbiota was revealed using redundancy analysis. ResultCompared with the normal group， the model group showed a prolonged sleep latency （P<0.05）， reduced sleep maintenance （P<0.01）， decreased central area activity time in the open field （P<0.01）， and reduced sugar preference rate （P<0.05）. Moreover， the model group also showed decreased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， decreased 5-HIAA/5-HT ratio （P<0.01）， downregulated mRNA expression of 5-HT1AR （P<0.01）， and upregulated mRNA expression of 5-HT2AR （P<0.05）. The proportion of Firmicutes decreased， while that of Bacteroidetes increased， leading to a decreased Firmicutes/Bacteroidetes （F/B） ratio （P<0.05）. Compared with the model group， the high-dose Huanglian Ejiaotang group exhibited a shortened sleep latency （P<0.01）， and increased sleep maintenance （P<0.01）. The low-dose Huanglian Ejiaotang group showed increased central area activity time （P<0.01） and an increased sugar preference rate （P<0.05）. The high-dose Huanglian Ejiaotang group exhibited increased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， increased 5-HIAA/5-HT ratio （P<0.05）， upregulated mRNA expression of 5-HT1AR （P<0.01）， and downregulated mRNA expression of 5-HT2AR （P<0.05）. The low-dose Huanglian Ejiaotang group displayed an increased proportion of Firmicutes and a decreased proportion of Bacteroidetes， resulting in an increased F/B ratio. At the phylum level， 5-HT， 5-HIAA， and MAO-A were positively correlated with Firmicutes and negatively correlated with Bacteroidetes. At the genus level， 5-HT， 5-HIAA， TPH， and MAO-A were negatively correlated with Prevotella and Lactobacillus and positively correlated with Blautia and Bacteroides. ConclusionHuanglian Ejiaotang can improve sleep deprivation-induced insomnia and depressive-like behavior by regulating the activity of the 5-HT system and the composition of gut microbiota.
ABSTRACT
Objective:To observe the clinical efficacy of addition and subtraction syndrome differentiation method of Huanglian Ejiaotang and Baihe Dihuangtang in the treatment of menopausal syndrome （MPS） and symptoms of deficiency of yin and hyperactivity of fire， and investigate its regulation effects on neuroendocrine network. Method:One hundred and forty-eight patients were divided into control group （74 cases） and observation group （74 cases） according to the random number table method. The patients in control group received estradiol/norethisterone acetate tablets， 3 mg/time， 1 time/day， while the patients in observation group received addition and subtraction of Huanglian Ejiaotang and Baihe Dihuangtang based on the same western medicine treatment as control group， orally 1 dose/day， with a treatment course of 3 months in both groups. Kupperman index （KI） score was evaluated before treatment and at 1 month， 2 months and 3 months after treatment. The Menopausal Quality of Life （MENQOL）， self-rating depression scale （SDS）， Self-rating Anxiety Scale （SAS）， Pittsburgh Sleep Quality Index （PSQI） and score of deficiency of yin and hyperactivity of fire were evaluated before and after treatment. The levels of serum estradiol （E2）， follicle stimulating hormone （FSH）， luteinizing hormone （LH）， norepinephrine （NE）， 5-hydroxytryptamine （5-HT）， nitric oxide （NO）， endothelin-1 （ET-1） and calcitonin gene-related peptide （CGRP） were detected before and after treatment. Result:The clinical efficacy in observation group was better than that in control group （Z=2.115， P<0.05）， and the KI scores in observation group at 1 month， 2 months and 3 months after treatment were lower than those in control group （P<0.01）， and the decrease of KI score in observation group after treatment was larger than that in control group （P<0.01）. The scores of various dimensions of MENQOL scale in observation group were lower than those in control group （P<0.01）. The scores of deficiency of yin and hyperactivity of fire， SAS， SDS and PSQI in observation group were all lower than those in control group （P<0.01）， and the level of E2 in observation group after treatment was higher than that in control group （P<0.01） ，while the levels of FSH and LH were lower than those in control group （P<0.01）.The levels of 5-HT and NE in observation group were higher than those in control group （P<0.01）.The NO level in observation group was higher than that in control group （P<0.01）， while the levels of CGRP and ET-1 were lower than those in control group （P<0.01）. During the study period， there were 5 drop-out cases， 2 excluded cases， and 67 completed cases in the control group. There were 7 drop-out cases， 1 excluded case， 66 completed cases in the observation group. Conclusion:On the basis of hormone replacement therapy， addition and subtraction of Huanglian Ejiaotang and Baihe Dihuangtang for patients with MPS （with symptom of deficiency of yin and hyperactivity of fire） can improve clinical symptoms， relieve depression and anxiety， improve sleep quality and enhance quality of life， and its clinical efficacy is superior to hormone replacement therapy alone. It can also regulate endocrine hormones， vasomotor factors and monoamine neurotransmitters， thereby improving clinical symptoms.
ABSTRACT
Objective:To observe the clinical efficacy of addition and subtraction therapy of Liuwei Dihuangwan combined with Huanglian Ejiaotang to primary insomnia in the elderly (liver and kidney Yin deficiency syndrome), and to investigate its regulation effect on neurotransmitter. Method:Randomly 136 patients were divided into control group (68 cases) and observation group (68 cases) by number table. Patients in control group got Estazolam tablets by oral administration before going to bed, 1-2 mg/time, 1 time/day. Patients in observation group got addition and subtraction therapy of Liuwei Dihuangwan combined with Huanglian Ejiao tang, 1 dose/day. Both groups of patients received sleep guidance and cognitive behavior guidance, with treatment course of 8 weeks. During the observation period, physical therapy and acupuncture could not been used. Before and after treatment, pittsburgh sleep quality index (PSQI), whole night polysomnography, sleep latency (SL), awakening times (AT), sleep efficiency (SE), rapid eye movement (REM), latency (RL), total actual sleep time (TST) and proportion of N1, N2, N3 and REM in the whole sleep stage were recorded. Scores of self-rating depression scale (SDS), self-rating anxiety scale (SAS), kidney Yin deficiency syndrome score and treatment emergent symptom scale (TESS) were graded, and levels of 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid(5-HIAA), gamma-aminobutyric acid (GABA), norepinephrine (NE) and dopamine (DA) were detected. Result:In rank sum test, clinical efficacy in observation group was better than that in control group (Z=2.115, PPPPPPPPχ2=9.945, PConclusion:Addition and subtraction therapy of Liuwei Dihuangwan combined with Huanglian Ejiaotang can improve sleep quality, prolong sleep time, alleviate depression, anxiety, and can also regulate neurotransmitters to improve sleep effect. The efficacy of PSQI is better than that of Estazolam tablets.