1.Study On The Mechanism Underlying The Effect Of CPAP On Arrhythmias In Patients With OSAHS,2.Study On The Mechanism Underlying The Effect Of NLRX1 On Soft Palate Tissue In Patients With OSAHS

Obstructive sleep apnea-hypopnea syndrome(OSAHS)is a widespread disease characterized by apnea and hypopnea,which is caused by complete or partial obstructions of the upper airway.These recurrent respiratory events lead to hypoxemia,hypercapnia,increased sympathetic nerve activity,significant fluctuations in intrathoracic pressure,sleep structural disorders and sleep fragmentation.OSAHS can cause diseases in all systems of the whole body,and increase the incidence and mortality of cardiovascular diseases.OSAHS has been shown to be significantly related to several cardiovascular conditions including coronary artery disease,hypertension,and congestive heart failure,as well as with arrhythmias.Many epidemiological studies have confirmed that there is a significant relationship between OSAHS and arrhythmias.About 58%of OSAHS patients have various types of arrhythmias,in which the incidence is much higher than that of non-OSAHS patients.Almost all types of arrhythmias can be observed in OSAHS patients.The incidence was positively correlated with the severity of OSAHS.With regard to the mechanism of arrhythmia in OSAHS patients,previous studies speculated that oxidative stress caused by nocturnal chronic intermittent hypoxia in OSAHS patients was one of the main causes of arrhythmia.Continuous positive airway pressure(CPAP)treatment is effective for the improvement of the symptoms of sleep apnea and is recognized as one of the therapeutic methods for OSAHS,To date,however,the relationship between oxidative stress and arrhythmias combined with OSAHS has been studied only sparingly.And there is still debate about the effect and mechanism of CPAP treatment on arrhythmia in OSAHS patients.Objective:The purpose of this study was to understand the general characteristics of arrhythmias in patients with OSAHS,and to compare the improvement of arrhythmias and the changes of biomarkers of oxidative stress in patients with OSAHS combined with arrhythmias before and after CPAP treatment,thereby evaluating the therapeutic effect and clarifying the mechanism of CPAP on arrhythmias in patients with OSAHS.Methods:Initially,107 patients were screened.After exclusion and matching design,a total of 64 were enrolled.All the 64 patients with OSAHS combined with arrhythmias were recruited from April 2014 to April 2017,with full clinical information.Patients referred to these clinics routinely underwent overnight polysomnography(PSG)and dynamic Holter monitoring.These 64 patients were composed of 38 males and 26 females,and the age ranged from 19 to 70 years old,with an average of 53 years old.All subjects in the study group were asked about their medical history in detail,including snoring,apnea,suffocation at night,open mouth breathing,dry mouth in the morning,nocturnal urine,morning headache and so on.Age,body mass index(BMI),neck circumference,waist circumference,blood pressure(before and after sleep),physical examination of otorhinolaryngology,years of OSAHS history were recorded,and epworth sleepiness scale(epworth sleepinessscale,ESS)was filled out.The 64 patients were divided into two groups according to the principle of paired design:32 patients in group A(control group),who were keeping unchanged their anti-arrhythmia treatment and 32 patients in group B(test group),who were subjected to unchanged pharmacological anti-arrhythmia therapy combined with CPAP.And each group consisted of 18 patients with severe OSAHS,8 patients with moderate OSA and 6 patients with mild OSAS.There were no significant differences in the type of arrhythmia and pharmacological treatment between the two groups.Before and after CPAP therapy,we compared the differences of AHI,LSp02,percent of total sleep time in stage N3(NREM 3)and stage R(REM),ESS and improvement of arrhythmias between the test group and the control group.The levels of native thiols,NADPH oxidase and MDA were measured simultaneously.Results:(1)In the 64 patients with OSAHS combined with arrhythmias,the main presenting symptoms were snoring in 63(98.44%),witnessed apneas in 34(53.13%),mouth breathing in 63(98.44%),nocturia in 49(76.56%),disturbed nocturnal sleep in 47(73.43%),morning headaches in 21(32.81%)and daytime sleepiness in 56(87.50%)with a mean ESS of 17.3.(2)The cardiac arrhythmia diagnosed in this cohort was represented by bradycardia(49),followed by sinus tachycardia(45),atrial tachycardia(29),atrial fibrillation(29,),premature atrial complex(26),premature ventricular complex(26),second-to third-degree atrioventricular block(19),ventricular tachycardia(17),sinus pause(16),paroxysmal atrial fibrillation(14)and premature junctional complex(12).(3)After 3 months of CPAP therapy,compared with the control group,AHI and ESS score were significantly decreased,percent of total sleep time in stage N3(NREM 3)and stage R(REM)was significantly increased,meanwhile,the lowest blood oxygen saturation was notably elevated.There were significant differences in these parameters between the two groups(P<0.05).(4)After 3 months of CPAP therapy,all types of arrhythmia were significantly reduced after CPAP treatment compared with drug treatment alone in patients with OSAHS(P<0.05).However,CPAP therapy was less effective for atrial fibrillation and paroxysmal atrial fibrillation than other types of arrhythmia,although the results were also statistically significant.(5)Before CPAP treatment,there were no statistically significant differences in the oxidative parameters between the two groups(P>0.05).CPAP therapy for 3 months significantly attenuated the oxidative stress.The serum native thiol was improved by CPAP treatment(P<0.05).The MDA and NADPH oxidase levels were significantly lower in the CPAP-treatment group than in the control group(P<0.05).Conclusion:(1)This study confirms that CPAP treatment effectively alleviates arrhythmias in patients with OSAHS.(2)This study confirms that CPAP treatment can improve the oxidative stress state of OSAHS patients,and the improvement of oxidative stress is related to the decrease of the incidence of arrhythmias.obstructive sleep apnea-hypopnea syndrome(OSAHS)is a disease characterized by recurrent upper airway collapse during sleep.It is characterized by recurrent hypoxemia and hypercapnia and sleep structural disorders,which in turn cause multiple system and multiple organ dysfunction.And it is an important risk factor for a variety of cardiovascular and cerebrovascular diseases.The main cause of OSAHS was the collapse of the upper airway during sleep.A sleep-stage electronic fiber laryngoscopy revealed that all patients with OSAHS had obstruction in the posterior soft palate.Therefore,soft palate lesions play an important role in the occurrence and development of OSAHS.The main cause of soft palate tissue lesions is tissue hyperplasia and fibrosis caused by chronic inflammation,so chronic inflammation of soft palate tissue is the main cause of OSAHS.After extensive review of the literature.We found that NLRX1,a members of the nucleotide binding domain and leucine-rich-repeat-containing proteins family(NLRs),has a significant negative regulatory effect on inflammatory responses induced by various factors(virus,lipopolysaccharide and others).It is a "brake" of overactivated inflammatory response,which may be closely related to chronic inflammation of soft palate in OSAHS patients.At present,there is no literature on whether NLRX1 protein is expressed in soft palate and how it is expressed,and whether NLRX1 has a regulatory effect on inflammatory response in soft palate,and how the mechanism underlying the effect of NLRX1 on soft palate tissue in patients with OSAHS.In this study,soft palate tissue samples were collected from patients with OSAHS and patients in the control group.The expression level of NLRX1 in soft palate tissues of OSAHS patients and the mechanism underlying the effect of NLRX1 on chronic inflammation in soft palate tissues were detected in this work by detecting NLRX1 and its signal molecules,inflammatory molecules and signal channels related to inflammation,so as to further understand the pathogenesis of OSAHS.It provides a new idea and basis for exploring the clinical treatment and prevention of OSAHS.Objective:The expression level of NLRX1 in soft palate tissues of OSAHS patients and the mechanism underlying the effect of NLRX1 on chronic inflammation in soft palate tissues were detected in this work.Methods:We selected patients with OSAHS(June 2017-June 2018)diagnosed by polysomnography(PSG)in Otorhinolaryngology and Sleep Medicine Center of Shandong Provincial Hospital affiliated to Shandong University.We conducted detailed screening to exclude OSAHS patients with a variety of other diseases(including chronic hepatitis,diabetes,rheumatic diseases,immune neuropathy,and coronary heart disease).A total of 18 eligible OSAHS cases were enrolled in this study.We divided patients into mild(6 cases),moderate(6 cases),and severe groups(6 cases)according to the severity of the disease.At the same time,we collected 6 patients with tonsillar benign tumors without a history of OSAHS as a control group.There were no significant differences in age and gender between the groups.All tissue samples were taken from the soft palate tissue about 1.0cm away from the free margin of the soft palate.The tissue was divided into 3 parts,each of which was about O.5cm*0.5cm*0.5cm.And one of which was fixed with paraformaldehyde,and the other immediately frozen in liquid nitrogen and stored in a-80 ? refrigerator.We further examined NLRX1 levels,inflammatory molecules(MCP-1,ICAM-1),major targets of pro-inflammatory signaling molecules(I?B?,NF-?B)and NLRX1 signaling pathways associated with viral infection(IRF3,IFN-?).To study the expression level and changes of NLRX1 in soft palate,and the role and mechanism of NLRX1 in chronic inflammation of soft palate,We used these experiments to study the expression and changes of NLRX1 in soft palate and the role and mechanism of NLRX1 in chronic inflammation of soft palate.Results:1.Immunohistochemistry confirmed that NLRX1 was expressed in the squamous epithelial basal layer and acanthosis cells of the soft palate tissue,and the positive expression was located in the cytoplasm.Compared with the control group,the level of NLRX1 gradually decreased with the severity of OSAHS(mild-moderate-severe).Therefore,the level of NLRX1 was significantly "negatively correlated" with the severity of OSAHS.2.Western blot and Real time-PCR showed that the inflammatory molecules(MCP-1,ICAM-1)in the soft palate of OSAHS patients were significantly up-regulated compared with those in the control group.And the inflammatory response was the most severe in patients with moderate OSAHS,and all the results were statistically significant.The results of Real time-PCR showed that I?B? was significantly down-regulated,while NF-?B was significantly increased.Western blot showed that I?B? was significantly down-regulated and p65NF-?B in nucleus was significantly increased.3.Further detection of anti-viral infection-related NLRX1 signaling channel molecules(IRF3,IFN-?),the results showed that.Compared with the control group,IRF3 and IFN-? were up-regulated in OSAHS patients.Although the amplitude of the increase was small,there were statistics significance.Conclusion:1.Our results indicate that NLRX1 is expressed in soft palate tissue,and the function and mechanism of NLRX1 has been broadened to the field of OSAHS for the first time.2.Compared with the control group,NLRX1 levels are gradually down-regulated with the severity of OSAHS patients(mild-moderate-severe).The inflammatory response in the soft palate tissue of OSAHS patients with different severity is significantly aggravated,and the inflammatory response is the most in soft palate tissue in patients with moderate OSAHS.3.NLRX1 is significantly "negatively correlated" with the expression of inflammatory and pro-inflammatory molecules.Down-regulation of NLRX1 in soft palate tissue in OSAHS patients relieves inhibition of inflammatory response,which leads to increased chronic inflammation,tissue hyperplasia,and fibrosis in soft palate,leading to airway obstruction.NLRX1-inflammation-airway obstruction-OSAHS may be an important path in the progression of disease in OSAHS patients.