A Study Of The Association Of Neurotransmitters And Inflammatory Cytokines In The Role Of Fatty Acids In Late Bedtime Cardiovascular Risk Factors

Objective By exploring the influence of neurotransmitters and inflammatory factors on the process of fatty acid-mediated cardiovascular diseases,the role of fatty acids and the pathophysiological basis of late bedtime affecting cardiovascular risk factors were explored,so as to provide guidance on sleep hygiene and diet management for the prevention of cardiovascular diseases and reduce the risk of cardiovascular diseases.Method This study adopted a convenient sampling method to collect samples from patients who visited the Second Affiliated Hospital of Xiamen Medical College from September 2021 to December2022.A total of 226 subjects were recruited for this study.General demographic information and clinical data were collected,and the current effective night bedtime was recorded.The subjects ranged in age from 35 to 87 years old,with an average age of（53.8±12.671）.There were 143 males and 83 females in total.Six patients with infectious diseases and nine subjects taking Melatonin（MT）were excluded,and 211 valid samples were included.The Pittsburgh sleep quality index（PSQI）was used to quantify the sleep condition of the subjects and calculate their bedtime at night.The research samples were grouped according to the first question of PSQI,namely,bedtime at night.They were divided into two groups:bedtime<11o’clock group（early bedtime group,n=126）and bedtime≥11 o’clock group（late bedtime group,n=85）.The general information of the samples in each group was investigated,and the subject‘s peripheral blood and cerebrospinal fluid（CSF）were collected.The levels of Orexin A（OXA）and MT in CSF,interleukin-10（IL-10）,and interleukin-18（IL-18）in serum,as well as Apolipoprotein A（Apo-A1）and Apolipoprotein B-100（Apo-B100）levels in plasma,were measured by Enzyme-linked immunosorbent assay（ELISA）.Free saturated fatty acid（SFA）and monounsaturated fatty acid（MUFA）in CSF and plasma were detected by Gas Chromatography（GC）.Age and Body Mass Index（BMI）were used as covariables to compare the differences in the dependent variable Apo-A1,Apo-B/Apo-A1 ratio and the differences in the contents of biomarkers IL-10,IL-18,OXA and MT between the two groups.The contents of 13 SFA and 5 MUFA were compared between the two groups.The correlation analyzed between bedtime,Apo-A1,Apo-B/Apo-A1,and biomarkers IL-10,IL-18,OXA,and MT were carried out in the two groups,respectively.And the correlation analyzed between bedtime,Apo-A1,Apo-B/Apo-A1,and different SFA and MUFA contents were carried out in the two groups,respectively.The results are conducted Bonferroni Correction;The common factors of 18 kinds of plasma free fatty acid（FFA）were extracted by dimensionality reduction factor analysis.A hierarchical stepwise linear regression analysis was performed for the factors and FAs correlated in the single factor analysis and the common factors of FFA extracted.Common factors of 18 kinds of CSF-FFA were extracted by dimensionality reduction factor analysis.Age,BMI,and the total score of Generalized Anxiexy Disorde-7（GAD-7）Patient Health Questionnaire-9（PHQ-9）were used as the covariable to conduct the binary logistic regression analysis of CSF-FFA and its common factor with bedtime.Results（1）The levels of dependent variables Apo-A1 and Apo-B/Apo-A1 differed between the two groups.The level of Apo-A1 in the early bedtime group was higher than that in the late bedtime group（P=0.014）,and the level of Apo-B/Apo-A1 was lower than that in the late bedtime group（P=0.012）.（2）The two groups had differences in OXA and IL-18 levels.OXA level in the early bedtime group was higher than that in the late sleep group（P<0.01）,and the IL-18 level was lower than that in the late bedtime group（P=0.041）.（3）The levels of palmitoleic acid and oleic acid in MUFA and palmitoleic acid,stearic acid,and arachidonic acid in SFA differed between the two groups.The above FA which different in the two groups,with its levels in the early bedtime group all higher than those in the late bedtime group(P_{palmitoleic acid}<0.01,P_{oleic acid}<0.01,P_{palmitoleic acid}<0.01,P_{stearic acid}<0.01,P_{arachidonic acid}=0.037).（4）Bedtime was not correlated with the dependent variables Apo-A1 and Apo-B/Apo-A1（P>0.05）,and the biomarkers Orexin A and MT,IL-10,IL-18 were not correlated with bedtime,Apo-A1 and Apo-B/Apo-A1（P>0.05）.Palmitoleic acid was negatively correlated with bedtime（P=0.01）.Palmitic acid was positively correlated with Apo-B/Apo-A1（P=0.036）.（5）Bedtime in the late bedtime group was negatively correlated with Apo-A1（P=0.034）,positively correlated with Apo-B/Apo-A1（P<0.01）,and positively correlated with OXA（P=0.04）;CSF MT was positively correlated with Apo-A1（P=0.036）,oleic acid,palmitic acid,and arachidic acid were positively correlated with Apo-B/Apo-A1(P_{oleic acid}=0.02,P_{palmitic acid}=0.018,P_{arachidic acid}=0.033).（6）Three common factors were extracted from 18 fatty acids in the early bedtime group,and five were extracted from 18 fatty acids in the late bedtime group.（7）Hierarchical stepwise linear regression results show that:In the early bedtime group,the independent variable bedtime had a statistically significant effect on palmitoleic acid（b=-4.124,t=-2.884,P=0.005）,and palmitoleic acid had a statistically significant effect on cardiovascular risk markers Apo-B/Apo-A1（b=0.000,t=2.541,P=0.012）.There was no statistical significance in the effects of the fatty acid common factor on Apo-A1 and Apo-B/Apo-A1（P>0.05）;In the late bedtime group,the effect of bedtime on Apo-A1,Apo-B/Apo-A1 was statistically significant(b=-0.074,t=-2.214,P_Apo-A1=0.03;b=0.123,t=2.902,P_Apo-B/Apo-A1=0.005),the independent variable bedtime had statistically significant effects on CSF Log OXA（b=0.038,t=2.442,P=0.017）.The effect of CSF MT on cardiovascular risk marker Apo-A1 was statistically significant(b=0.001,t=2.665,P_Apo-B/Apo-A1=0.009).The effects of palmitic acid on cardiovascular risk markers Apo-B/Apo-A1 were statistically significant（b=0.001,t=2.805,P=0.006）.Fatty acid common factor 1（composed of oleic acid,palmitoleic acid,palmitic acid,heptadecanoic acid,stearic acid,and carbonic acid）in the late bedtime group had statistically significant effects on Apo-B/Apo-A1（b=0.096,t=2.449,P=0.017）.（8）Five common fatty acid factors were extracted from the CSF-FFA of the whole sample.（9）Binary logistic regression showed that myristate acid and tricosanoic acid in CSF fatty acids reduced the probability of late bedtime(OR=0.648,95%CI 0.485-0.865,P_{myristate acid}=0.003;OR=0.767,95%CI 0.631-0.934,P_{tricosanoic acid}=0.008);Eicosanoic acid and tetracenoic acid increased the likelihood of late bedtime(OR=1.299,95%CI 1.021-1.652,P_{Eicosanoic acid}=0.033;OR=1.142,95%CI 1.006-1.298,P_{Tetracenoic acid}=0.041);There were no significant differences in the effects of CSF-FFA on Apo-A1 and Apo-B/Apo-A1（P>0.05）.Conclusion（1）Bedtime late is an essential factor that leads to increased cardiovascular risk.When bedtime is after 11 pm,the last going to bed,the higher the risk of cardiovascular disease.FA has an essential mediating role;a high level of palmitic acid is a significant risk factor for cardiovascular disease regardless of whether bedtime occurs before or after 11 o’clock.However,when bedtime is late,palmitic acid and oleic acid not only affects cardiovascular health alone but also their combined effects with palmitic acid,heptadecanoic acid,stearic acid,and carbonic acid are also significant risk factors for cardiovascular risk,and their impact degree higher than that of palmitic acid alone.Therefore,excessive SFA and animal-source MUFA should be avoided as far as possible.Balance and variety of diet should be paid attention to while adjusting sleeping habits;（2）Low levels of MT are an essential risk factor for cardiovascular disease,and it is worth noting that late bedtime plays a crucial regulatory role;（3）OXA level is also increased under the influence of late bedtime behavior.Although it does not directly affect cardiovascular disease,it promotes the imbalance of fatty acid intake by inducing increased appetite,thus affecting the occurrence and development of cardiovascular diseases.Interestingly,long-term late bedtime will reduce OXA levels,presenting a state of short-term increase and long-term consumption.（4）High levels of CSF myristate acid and tricosanoic acid can increase the possibility of early sleep.Higher CSF eicosanoic acid and tetracenoic acid levels can promote late sleep behavior.Therefore,high levels of SFA can increase triglyceride levels,thus leading to hypoxia of blood and sticky brain tissue,which is more likely to cause sleepiness.Therefore,when adjusting sleep habits,attention should also be paid to avoiding excessive intake of single,SFA-rich foods and paying attention to the balance of the diet.