| Insomnia is a subjective experience characterized by a diffculty in falling asleep and/or staying asleep,which results in the failure of sleep quality or quantity to meet an individual's normal physiological needs and affects their social functions.It is the most common sleep disorder with very high rates of morbidity.According to an investigation by the World Health Organization,?1/3 of the world's population suffers from sleep disorders.The percentage of people with various types of sleep disorders in China is signifcantly higher than the rest of the world(27%).Thus,effective prevention and treatment of insomnia is a main focus of research in the worldIn clinical treatment,hypnotics are primarily used in modern medicine.This drug treatment may be accompanied by many side effects.It is not an ideal therapeutic regimen due to its poor longterm effcacy and addiction,and tolerance,in the case of long-term administration.Mongolian medicine has a unique mechanism and good effcacy in treatment of insomnia.In Mongolian medicine,it is believed that the imbalance among Heyi,Xila,and Badagan,predominance of Heyi in the heart and white meridian,and Heyi blood intermingling arising from dysfunctions are the basic etiology and pathogenesis.Such negative emotions as tension,worry,fear,depression and anxiety;and social environment,diet,daily life and movement conditions are external factors.In the Mongolian medicine,the treatment of insomnia mainly focuses on relieving Heyi and regulating Heyi,Xila,and Badagan.The warm needling therapy is one of most common methods for the treatment of insomnia in Mongolian medicine as it functions in warming and smoothing meridians,regulating qi and blood,regulating voxels,enhancing immunity,and preventing and treating diseases.It is widely accepted way of treatment by the patients due to such characteristics as high effciency,safety,no side effects,simplicity and no drug dependence.Thus,the warm needling therapy has considerable advantages in the treatment of insomnia,such as signifcant effcacy,safety,simplicity,and economy.This topic in Mongolian Medical for PCPA cause insomnia rats model of hypnosis as the breakthrough point,insomnia rats behavior related indicators,from the level of genes related to Warm Acupunture before and after treatment of miRNAs and target genes were discussed,at the same time of insomnia related cytokines,observe the neurotransmitter content,explore the Mongolian Medical Warm Acupunture biological basis for the treatment of insomnia,preliminary clarify Mongolian Medical Warm Acupunture mechanism of multilayer complex.ObjectiveOur study frst investigates the role of the Mongolian medical warm acupuncture in hypnotizing the PCPA-induced insomnia models.We also discuss the levels of related microRNAs and target genes before and after warm acupuncture.Finally,we observed the insomnia-related cytokines and neurotransmitter content,probed into the biological foundation of Mongolian medical warm acupuncture in treating insomnia in combination with the clinical tests,and preliminarily explain the multi-dimension complex mechanism of the Mongolian medical warm acupuncture.MethodsA total of 90 clean Sprague-Dawley rats.were then intraperitoneally injected with PCPA.The Sprague-Dawley rats were divided into 6 groups randomly,the normal ?,normail?,model,warm acupuncture,warm acupuncture with model,and medication with model groups of 15 rats each.2.Determination of the differential expression profile of microRna in the brain tissue of the insomnia rats before and after Mongolian medical warm acupuncture analysis of the microRna expression pro file.3.Use the enzyme-linked immunosorbent assay(ELISA)to detect the levels of the interleukins of IL1,IL2,and IL-6 and the tumor necrosis factor-a(TNF-a)at the above sites.Detection of monoamine neurotransmitters in the brain tissue was conducted using the high performance liquid chromatography-electrochemical detection method to determine the levels of noradrenaline(NE),dopamine(DA),5 hydroxytryptamine(5HT),glutamic(Glu)acid,and ?aminobutyric acid(GABA).Result1.In the amount of Sleep,Mongolian medicine Warm Acupuncture group compared with the blank group of prolonged sleep,have obvious difference(P<0.01);Mongolian medicine temperature on rats sleep rate has no effect,no significant difference compared with the blank group(P>0.05);Incubation period of sleep has no obvious effect,no significant difference compared with the blank group(P>0.05).2.In OFT experiment,each group rats insomnia on the seventh day,on the activity time model group was obviously higher than that of blank group(P<0.01);The retention period of model group is higher than the blank group,but no significant difference(P>0.05).Number of stand,stand number of model group was obviously higher than that of control group,there are significant differences(P<0.05);Mongolian medicine group needle temperature was lower than that in model group(P<0.05).3.The rats receiving Mongolian medicine warm acupuncture had 141 miRNAs with differential expression compared with the normal rats.The expression level of miR-101a in the cells of the hippocampus of the insomnia rats transfected with miR-101a mimics increased significantly at 72 h(P<0.05).The activity of the neuronal cells transfected with miR-101a inhibitor increased significantly at 72 h(P<0.05).4.The western blotting result indicated that the expression of the PAX8 protein in the neuronal cells of the insomnia model rats was inhibited and downregulated significantly at 72 h after addition of miR-101a mimics compared with that in the scramble added group(P<0.01).5.The levels of the interleukins IL-1,IL-2,and IL-6 and the tumor necrosis factor-a in the hypothalamus,hippocampus,and prefrontal cortex decreased significantly compared with those in the blank control group(P<0.05).The levels of noradrenaline,dopamine,and glutamic decreased significantly following warm acupuncture or western medicine treatment(P<0.05).Conclusion1.Mongolian Medicine Warm Acupuncture can reduce the spontaneous activity,improve behavioral effects as reaction.2.Mongolian medicine Warm Acupuncture can directly increase insomnia rats miR-101-a level,and verified the Mongolian medicine temperature adjustable needle to improve insomnia and miR-101 a PAX8 is directly related genes.3.Indicate that warm acupuncture had significant effcacy in treatment of insomnia rats.Its basic effcacy was equivalent to that of the drug treatment.4.1n conclusion,this study demonstrates that the upregulation of miR-101a in the rats treated with warm acupuncture is directly associated with PAX8 regulation. |
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