| Background:Sleep disorder is not only one of the risk factors for the occurrence and development of Alzheimer’s disease(AD),but also the most common accompanying symptoms.To improve sleep as another way to treat dementia is a new idea of prevention and treatment of AD.However,western medicine treatment of sleep disorders in AD has poor efficacy and side effects.The method of clearing heat and resolving phlegm in traditional Chinese medicine is an effective method for the treatment of cognitive decline,sleep disorders and other psychobehavioral symptoms in the middle and late stages of ad found by our team in long-term clinical research.However,the mechanism of clearing heat and resolving phlegm in traditional Chinese medicine is still unclear.Objectives:The purpose of this study is to study the characteristics of AD sleep-wake rhythm,and explore the regulatory effects and possible mechanisms of Chinese medicine for clearing heat and resolving phlegm on AD sleep-wake rhythm and cognition from the perspective of clock genes and neuroinflammation.Methods:The study is divided into a clinical part and an experimental part.The clinical study is an observational study.The characteristics of sleep disorders in AD patients are analyzed by a wrist body movement instrument.Experimental research evaluates the therapeutic effects and possible mechanisms of Chinese medicines for clearing away heat and resolving phlegm.1.Sleep-wake rhythm characteristics of patients with Alzheimer’s disease:According to the inclusion and exclusion criteria,44 subjects were divided into cognitively normal group(n=14)and mild cognitive impairment(MCI)group(n=18)and the AD group(n=12),all subjects received neuropsychological assessments,and at least 5 consecutive days in a normal living environment to monitor sleep-wake status by wearing a wrist-type body exercise device.Compare the sleep parameters between the three groups:time in bed(TIB),total sleep time(TST),time to fall asleep(SOL),sleep efficiency(SE),wake-up time after falling asleep(WASO),and analyze the sleep of AD patients and MCI patients The correlation between parameters and cognition(MMSE,CDR,DSR)and function(ADL).2.The regulation of heat-clearing and phlegm Chinese medicine on sleep and cognition:10-month-old APP/PS1 double transgenic mice were randomly divided into model group,western medicine group,and traditional Chinese medicine group.Normal wild-type C57/BL6J was used as the normal control group.The western medicine group was given donepezil,the traditional Chinese medicine group was given the heat-clearing and phlegm Chinese medicine,the model group and the control group were given an equal volume of 0.5%CMC,once a day by gavage for 3 months.(1)The small animal activity monitor was used to observe the circadian rhythm changes of the autonomous activities of the mice in each group after the intervention.Western blot(WB)technology was used to detect the protein expression of the clock gene BMAL1 in the hippocampus,and to evaluate the effect of the traditional Chinese medicine for clearing heat and resolving phlegm on AD model mice.The influence of sleep and circadian rhythm.(2)Morris water maze experiment was used to detect the behavioral changes of mice after intervention,and WB was used to detect the content of Aβ1-42 in hippocampus to evaluate the effect of Chinese medicine for clearing away heat and removing phlegm on behavior and pathological proteins in AD mice.(3)Immunohistochemistry was used to detect the expression of astrocyte activation marker glial fibrillary acidic protein(GFAP)in the paraffin sections of the hippocampus of each group of mice,and Real-time PCR was used to detect the levels of TNF-α and COX2 in the hippocampus.The effect of Chinese medicine for clearing away heat and removing phlegm on the activation degree of astrocytes and neuroinflammation.Results:1.AD patients have sleep-wake disorders:(1)The awakening time(62.20±37.71)minutes of AD patients after falling asleep tends to be higher than that of the normal cognitive elderly(47.50±15.00)minutes,and the fragmentation of sleep is more serious There is no significant difference in bed time,total sleep time,sleep latency,and sleep efficiency in AD patients,MCI patients and the elderly with normal cognition.(2)A subgroup analysis of AD patients according to gender showed that female AD patients spent significantly longer time in bed than males(P=0.033),but there were no significant differences in total sleep time,latency to fall asleep,sleep efficiency,and wake-up time during sleep.According to the subgroup analysis based on the degree of cognition,there was no significant difference in the results of the sleep parameters of patients with mild and moderate AD.The subgroup analysis was performed according to the time of awakening after falling asleep.With 60 minutes as the cutoff value,there was no significant difference between the two groups of patients in cognition and ability of daily living,but the age of AD patients greater than 60 minutes was significantly older than that of patients less than 60 minutes(P=0.003).(3)The degree of sleep disorder in AD patients has a certain correlation with cognition and function.There was a negative correlation between SOL and MMSE(r=-0.619,P<0.05).SE was negatively correlated with ADL(r=-0.649,P<0.05).TIB,TST,WASO have no correlation with cognition and function.There is no correlation between sleep parameters and cognition and function in MCI patients.(4)In the light environment,the mice in the control group of 7 months and 10 months(adult)are more active than the AD model mice,but at the age of 13 months(old age),the model group has more activities than the control group;dark environment The 7-month-old,10-month-old,and 13month-old model groups were more active than the control group,but there was no statistical difference between the groups.2.Chinese medicine for clearing heat and phlegm can regulate sleep-wake disorders in APP/PS1 mice and improve learning and memory:(1)Chinese medicine for clearing heat and phlegm improves the level of BMAL1 in APP/PS1 model mice,thereby optimizing sleep in AD mice-Awakening rhythm.(2)Chinese medicine for clearing away heat and resolving phlegm can improve the learning and memory ability of model mice,especially the ability of space exploration.After the intervention of traditional Chinese medicine,the staying time in the target quadrant of mice is significantly longer than that of the model group(P<0.05),and the number of crossing platform quadrants(5.21)±2.58)times better than the model group(3.44±2.13)times,but there is no statistical difference.In terms of escape latency,compared with the model group,there was no significant difference in the escape latency between the traditional Chinese medicine group and the western medicine group in the positioning navigation experiment.(3)Chinese medicine for clearing heat and resolving phlegm can reduce the deposition of pathological protein Aβ1-42 in the hippocampus of AD mice.3.Chinese medicine for clearing away heat and resolving phlegm can inhibit the excessive activation of astrocytes and reduce neuroinflammation:(1)Pathological examination found that the hippocampal GFAP expression of AD mice after the intervention of traditional Chinese medicine was significantly reduced(P<0.05),and astrocytes Excessive activation is suppressed.(2)The content of TNF-α and COX2 in hippocampus tissue was significantly reduced by Real Time-PCR(P<0.05),and neuroinflammation was alleviated.Conclusions:(1)AD patients may attenuate sleep-wake rhythm and increase fragmented sleep.(2)Chinese medicine for clearing heat and resolving phlegm can improve the sleep-wake rhythm of AD model mice,which may be related to increasing the expression of clock gene Bmall,(3)Chinese medicine for clearing away heat and resolving phlegm may improve the learning and memory ability of AD mice by reducing the deposition of Aβ1-42 pathological protein;inhibiting the excessive activation of astrocytes and reducing the neuroinflammatory response.(4)The Chinese medicine for clearing heat and resolving phlegm may increase the expression of the circadian clock gene Bmall to regulate the activation of astrocytes and neuroinflammation,reduce the deposition of Aβ1-42 pathological protein,and effectively protect neurons.(5)Chinese medicine for clearing away heat and resolving phlegm may improve cognition by optimizing sleep.Due to the small number of patients and the short duration of traditional Chinese medicine intervention,the above conclusions need to be further studied and verified. |
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