| Objectives:Through analyzing the mitochondrial function changes and damage factors of asthma patients with different severity and viscera deficiency,try to find the key link of the mitochondrial respiratory chain inhibition and dysfunction,reveals L-Arg/ADMA metabolism mechanism in mitochondria injury,oxygen metabolism,pathogenesis of asthma.From metabolism related substance-chain,which can reduce oxidative damage by regulating the metabolism of the body,to find a new target for the prevention and treatment of asthma.Methods:According to the diagnostic criteria of the 2016 GINA program,122 cases of asthma patients who met the diagnostic criteria were selected,and the clinical data were collected,including gender,age,BMI,smoking and allergic rhinitis,pulmonary function classification,and viscera deficiency.According to the diagnostic criteria of severity of asthma were divided into intermittent state(Level 1),mild asthma(Level 2),moderate asthma(Level 3),severe asthma(Level 4),according to the deficiency of Zang Fu syndrome diagnosis standard into lung qi deficiency group,spleen and lung,lung spleen and kidney deficiency group and 44 cases of normal control group.Blood samples were collected(18-20ml),using high performance liquid chromatography combined with mass spectrometry(HPLC-MS/MS)detection of L-arginine(L-Arg)and ADMA;detect-ing PRMT1,DDAH1,NO,ROS,TBARS,8-iso,LPO;Mitochondrial ultrastructure of platelet mitochondria was observed by trans-mission electron microscope,flow cytome-try and JC-1 method for the determination of platelet mitochondrial membrane potential(△Ψm),ATP and COX were detected to evaluate the function of mitochondria.The patients’ case data were set up by Excel form,and the difference and correlation between them were analyzed by SPSS 17.0 statistical software.The general data of patients,mito-chondrial function,arginine and related metabolic indexes were analyzed by descriptive analysis,the asthma group and the control group compared with t test;different viscera deficiency group and different severity groups when compared with normal controls using the one-way ANOVA,then the LSD test;using constituent ratio to describe severity distribution proportion in viscera deficiency.Pearson correlation analysis was used to analyze the relationship among the indexes,and the difference was statistically significant in P<0.05.Results:1)In 122 cases of asthma patients,41 cases is male,81 cases is female,age distribution of patients at 17-65 years old,mean age 48.28±10.61.In 44 cases of normal group,29 cases is male,15 cases is female,age distribution in 17-64 years old,mean age 49.50±10.70.The comparison between the asthma group and the healthy control group,there is no significant difference in sex ratio and age distribution(P>0.05).Comparison of age in different organs deficiency groups,the difference was statistically significant(P<0.05),lung qi deficiency group,lung spleen deficiency group had difference with lung spleen and kidney deficiency group(P<0.05).Comparison of pulmonary function with FEV1,FEV1/FVC,PEF between different viscera deficiency group and normalgroup,the difference was statistically significant(P<0.05),there were no significant differences between different organs deficiency in patients with asthma(P>0.05).There was no significant difference in smoking and allergic rhinitis between asthmatic patients with different organs deficiency(P>0.05).The age difference of asthma patients with different severity was not statistically significant(P>0.05).There was no significant difference in BMI between patients with different severity of asthma(P>0.05).There was no significant difference in smoking and allergic rhinitis(P>0.05).2)the plasma levels of 8-iso,TBARS,ROS,LPO and MDA in asthmatic patients were higher than normal group,and the differences were statistically significant(P<0.05).The MMP,ATP and COX were statistically significant difference between asthma group and the normal group(P<0.05),asthma group decreased compared with the normal group.No significant difference in plasma ADMA in asthma group and normal control group(P>0.05);Compared with normal group,L-Arg decreased(P<0.05);comparison of L-Arg/ADMA,asthma group was lower than normal group(P<0.05).No significant differences in PRMT1 between asthma group and normal group(P>0.05);DDAH1 of asthma group was lower than normal group(P<0.05).MMP was negatively related to LPO(r=-0.1983 P<0.05),ATP and COX was negatively correlated with 8-iso(r=-0.193,r=-0.423,P<0.05),and COX was negatively correlated with ROS(r=-0.141,P<0.05).ATP was negatively correlated with ADMA(r=-0.312,P<0.05),was positively correlated with L-Arg/ADMA(r=0.236,P<0.05),COX was negatively correlated with ADMA(r=-0.269,P<0.05),and was positively correlated with DDAH1(r=0.334,P<0.05).There was no significant correlation between NO and mitochondrial function index(P>0.05).8-iso and ADMA were positively related(r=0.377,P<0.05),and negatively correlated with L-Arg/ADMA(r=-0.330,P<0.05),negatively correlated with plasma DDAFH1 levels(r=-0.224,P<0.05),no significant correlated with L-Arg(P>0.05).TBARS and ADMA were positively related(r=0.296,P<0.05),and negatively correlated with L-Arg/ADMA(r=-0.184,P<0.05),negatively correlated with DDAH1(r=-0.182,P<0.05),no significant correlated with L-Arg(P>0.05).There was a positive correlation between NO and 8-iso(r=0.196,P<0.05).There was a negative correlation between ADMA and DDAH1(r=-0.224,P<0.05),which was not significantly correlated with L-Arg and PRMT1(P>0.05).There was no correlation between NO and L-Arg and L-Arg/ADMA(P>0.05).LPO was negatively correlated with PEF(r=-0.215,P<0.05);MDA was negatively correlated with FEV1,FEV1/FVC,PEF(r=-0.409,r=-0.325,r=-0.360,P<0.05);ROS was negatively correlated with FEV1,FEV1/FVC,PEF(r=-0.415,r=-0.23,r=-0.341,P<0.05).Age,sex,BMI and oxidative stress were taken as independent variables(X).FEV1%and PEF were taken as dependent variables(Y).Multilinear regression analysis and regression equation were constructed.The results showed that plasma levels of malondialdehyde(MDA),reactive oxygen species(ROS)and body mass index(BMI)were all shadow.The FEV1%risk factors(Y=163.305-0.958 MDA-0.016 ROS-0.685 BMI,F=26.818,R2=0.601,P<0-001)in patients with asthma were higher fitting,which could explain 60.1%FEV1%of the results.The plasma levels of malondialdehyde(MDA)and reactive oxygen species(ROS)were all risk factors for PEF in patients with asthma(Y=141.854-0.5).63MDA-0.008ROS,F=91.435,R2=0.693,P<0.001.The equation results show that the fitting degree is high,which can explain 69.3%PEF results.3)Comparison of different severity of asthma patients and normal group,the plasma 8-iso,the difference was statistically significant(P<0.05);There was no significant difference between the plasma of the patients with different severity of the disease and the normal group in TBARS(P>0.05).Comparison of plasma MDA,ROS in different severity of asthma patients and normal group,the difference was statistically significant(P<0.05),higher than normal group(P<0.05),plasma MDA,ROS is higher in severe asthma than in intermittent state and mild asthma(P<0.05).The difference of LPO between different severity of asthma group and normal group was statistically significant(P<0.05).Comparison of MMP,ATP and COX between different severity of asthma group and the normal group,there were significant differences(P<0.05),lower than the normal group,comparison of L-Arg,L-Arg/ADMA,DDAH1 between different severity gourp and normal group,there were significant differences(P<0.05),lower than normal.No statistically significant difference between different severity groups of asthma(P>0.05).Comparison of NO between different severity gourp and normal group,there were significant differences(P<0.05),the different severity of the disease groups were higher than normal(P<0.05).There was no significant difference in plasma ADMA and PRMT1 between different severity asthma groups and normal group(P>0.05).The age,BMI and oxidative stress of asthmatic patients were used as independent variables(X)and severity grading(Y)for logistic regression analysis.The predictive model was established.The results showed that plasma levels of malondialdehyde(MDA),reactive oxygen species(ROS)and age were risk factors affecting the severity of asthma.Logit P=-6.861+0.118 MDA+0.003 ROS+0.044 years old,by likelihood ratio test,χ2=43.403,P<0.001,equation model has statistical significance;by Hosmer-Lemeshow goodness of fit test,χ2=7.070,P>0.05)。4)Comparison of 8-iso between viscera deficiency asthma groups and normal group,the difference was statistically significant(P<0.05),the asthma group was significantly higher than the normal group(P<0.05).In different viscera deficiency groups,there were no significant statistical differences(P>0.05).There was no significant difference in TBARS between the viscera deficiency groups and normal group(P>0.05).Comparison of LPO and MDA in viscera deficiency groups and normal group,the differences were statistically significant(P<0.05),the viscera deficiency asthma groups was significantly higher than normal group(P<0.05).Comparison of ROS in viscera deficiency asthma groups and normal group,there was no statistically significant difference in lung qi deficiency group and the normal group(P<0.05),but the lung and spleen deficiency group,lung spleen and kidney deficiency group were significantly higher than the normal group(P<0.05).Comparison of MMP,ATP and COX in viscera deficiency asthma groups and normal group,there were significant differences,lower than normal group(P<0.05),but the different viscera deficiency asthma groups was no statistically different with each other(P>0.05).Comparison of L-Arg,L-Arg/ADMA,DDAH1 in different viscera deficiency asthma groups and normal control group,there were significant differences(P<0.05),lower than normal,but the different viscera deficiency asthma groups was no statistically different with each other(P>0.05).There was a significant difference in plasma NO between asthmatic patients with different viscera and normal(P<0.05),which was significantly higher than normal.There was no significant difference in plasma ADMA and PRMT1 between asthmatic patients with different viscera and normal control group(P>0.05).The age,lung function(FEV1%,PEF)and oxidative stress of asthmatic patients were used as independent variables(X)and viscera deficiency status(Y)for logistic regression analysis.The predictive model was established.The results showed that age and PEF of asthmatic patients were risk factors for viscera deficiency status(Logit P=-0.871+0.173age-0.167 PEF,by likelihood ratio test,χ2=102.801,P<0.001,the equation has statistical significance;Hosmer-Lemeshow goodness of fit test,χ2=6.701,P>0.05),the age of asthmatic patients and PEF can be used to predict the development trend of visceral deficiency.Conclusion:1)It is confirmed that mitochondrial dysfunction is an important factor in the pathogenesis of asthma.It is found that abnormal DDAH/L-Arg/ADMA pathway is an important link in the pathogenesis of asthma.The deficiency of DDAH1 and L-Arg caused ratio of L-Arg/ADMA decreased and ADMA relative surplus in patients with asthma,Which leading to mitochondrial respiratory chain inhibition and mitochondrial dysfunction,and the metabolism of active oxygen,oxygen accumulation and promote oxidative damage--this is the key of asthma.It is clearly pointed out that the lower the DDAI-I1 and L-Arg/ADMA,the more severe the mitochondria damage is,the more the reactive oxygen species accumulate.2)We may reduce the excessive expression of mitochondria damage and active oxygen accumulation through keeping the L-Arg/ADMA ratio balance by increasing L-arginine or regulating the DDAH1-ADMA,but it still need further research.3)It is suggested that mitochondrial damage in asthmatic patients may also be an important reason for over expression of NOS and the production of NO.When mitochondria was injured,respiratory chain was inhibited,a large number of Active Oxygen was produced and caused epithelial cell injury,airway inflammatory cell aggregation and activation,a large number of inflammatory cytokines and inflammatory mediators were released,iNOS expression increased significantly in endothelial cells,smooth muscle cells and macrophages that a large number of NO was produced.But it needs further research.4)It was showed a consistent tendency in viscera deficiency,clinical condition of patients with asthma,oxygen metabolism and progress of oxidative injury.Theory of TCM viscera deficiency in some extent may be external representation of the changes in metabolism. |
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