To Investigate The Correlation Between The Clinical Features And Intestinal Flora,TNF-α And IL-8 Levels Of Parkinson’s Disease

Objective: To investigate the correlation between the clinical characteristics of different syndrome types in patients with Parkinson’s disease(PD)and the abundance and types of intestinal flora as well as the levels of serum inflammatory cytokines TNF-α and IL-8,and to provide new ideas for the treatment and research of different syndrome types in traditional Chinese medicine of Parkinson’s disease.Method:Study 1: A total of 90 patients with primary Parkinson’s disease who visited inpatient department and outpatient department of Neurology Department of our hospital from January 2022 to February 2023 were selected as the experimental group.General information such as gender,age and medical history were collected according to inclusion and exclusion criteria,and TCM clinical syndrome differentiation was conducted according to the syndrome scale of Parkinson’s disease,which was divided into 5categories according to common clinical TCM syndrome types.The five syndromes were liver-kidney Yin deficiency,liver-wind internal movement,qi and blood deficiency,kidney deficiency and pulpia reduction,and phlegm turbidity.The Parkinson’s Disease non-motor Symptom Evaluation Scale(NMSS),Parkinson’s Disease Sleep Scale(PDSS),Hamilton Depression Scale(HAMD),and MDS-UPDRS Part III scale were scored for each enrolled patient.To compare the difference of clinical features among patients with different syndromes.Study 2: A total of 90 patients with primary Parkinson’s disease who met the criteria in experiment 1 were taken as the experimental group and 30 healthy people without PD as the control group.The blood of the two groups of subjects was collected and the serum concentrations of tumor necrosis factor(TNF-α)and interleukin-8(IL-8)were detected,and the correlation between different syndrome types and serum concentrations of TNF-α and IL-8 as well as clinical characteristics was analyzed.Study 3: In this study,30 stool samples of patients with primary Parkinson’s disease were selected as the experimental group,27 stool samples were qualified and effective,20 stool samples of non-PD healthy people were selected as the control group,19 stool samples were qualified.A total of 46 stool samples of effective subjects were collected,and microbial population abundance in feces was determined by 16S-r DNA sequencing technology for analysis.The differences of intestinal flora abundance between PD patients and normal people were compared,and the differences of liver wind internal movement syndrome(12 cases)and sputum turbidities(8 cases)in PD patients were compared.The correlation of dominant flora abundance with clinical features,TNF-α and IL-8 in PD group was compared.Results:Study 1: Clinical characteristics and TCM syndrome analysis of PD patients: 1.The proportion of syndrome differentiation types from high to low was successively distributed as deficiency of liver and kidney Yin,internal movement of liver wind,deficiency of qi and blood,turbidness of phlegm,deficiency of kidney and pulp.2.Comparison of NMSS score among five TCM syndrome groups: Kidney deficiency and pulp reduction(114.13±28.93)> liver and kidney Yin deficiency(97.69±46.69)> liver wind internal movement(72.18±32.84)> phlegm turbidity internal accumulation(60.00±22.03)> Qi and blood deficiency(68.61±20.17),the results were statistically significant(P<0.05;P=0.05),the other scales had no correlation among the five syndrome groups.3.Correlation analysis within each syndrome group: Liver and kidney Yin deficiency was positively correlated with NMSS score,HAMD score and UPDRS-III score(P<0.05),and negatively correlated with PDSS score(P<0.05).Qi and blood deficiency were positively correlated with HAMD score(P<0.05).The decrease of kidney deficiency pulp was positively correlated with NMSS score and HAMD score(P<0.05),but negatively correlated with PDSS score(P<0.05).Study 2:1.Comparison of IL-8 and TNF-α levels between normal subjects and subjects with 5 syndrome types was statistically significant(P<0.05;P<0.05).After pairwise comparison,the IL-8 level of normal people and liver wind internal movement was lower than that of liver and kidney Yin deficiency,kidney deficiency and pith reduction(P<0.05).The IL-8 level of kidney deficiency and pulp reduction was higher than that of liver and kidney Yin deficiency,qi and blood deficiency and phlegm turbidity(P<0.05).The levels of TNF-α in normal people were lower than those of liver wind internal movement,liver and kidney Yin deficiency,qi and blood deficiency,kidney deficiency and pulp reduction and phlegm turbidities(P<0.05).Study3:Study 3: 1.At the level of the phyla family,PD and healthy people in Firmicutes,Actinobacteria,Bacteroidetes,Bacilli,Clostridia,Bacteroidia,Clostridiales,Lactobacillales,Bacteroidale The abundance of s,Lactobacillaceae,Prevotellaceae and Bacteroidaceae had statistical significance.2.Actinobacteria and Clostridia were positively correlated with NMSS(P<0.05);Firmicutes,Actinobacteria,Bacteroidetes and Clostridia were correlated with PDSS(P<0.05).Actinobacteria were positively correlated with HAMD(P<0.05).The Actinobacteria were positively correlated with UPDRS-III(P<0.05).3.TNF-α was positively correlated with Firmicutes,actinobacteria and Clostridia(P<0.05);it was negatively correlated with Bacteroidetes(P<0.05).There was no correlation between IL-8 and the relative abundance of intestinal flora in PD.4.Roseburia may be related to liver wind internal motion syndrome;the turbidities of phlegm may be related to the abundance of microflora in Verrucomicrobia,Verrucomicrobiae,Verrucomicrobiales,and Verrucomicrobiaceae.Conclusion:Study 1:1.TCM syndrome differentiation of 90 PD patients was mainly liver and kidney Yin deficiency syndrome 2.NMSS showed statistical difference in the five TCM syndromes,and the other scales showed no correlation between the five syndrome groups.3.Yin deficiency of liver and kidney was positively correlated with NMSS,HAMD and UPDRS-III,but negatively correlated with PDSS.Deficiency of qi and blood was positively correlated with HAMD.The decrease of kidney deficiency was positively correlated with NMSS and HAMD,but negatively correlated with PDSS.Study 2:1.IL-8 level was higher than TNF-α level.2.There was statistical significance in IL-8 and TNF-α between normal subjects and subjects with 5 syndrome types;the concentration of IL-8 was the highest in the syndrome of kidney deficiency and pulp reduction.The level of TNF-α in normal subjects was lower than that in 5 syndrome types.3.There was no significant difference between the syndrome and IL-8 and TNF-α levels.NMSS,PDSS,HAMD and UPDRS-III were positively correlated with TNF-α,but not with IL-8.Study 3: 1.At the level of phylum,class,order and family,there were statistical differences in the relative abundance of intestinal flora between Parkinson’s disease patients and healthy people.2.Actinobacteria and Clostridia were positively correlated with NMSS;Firmicutes,Actinobacteria,Bacteroidetes and Clostridia were correlated with PDSS.Actinobacteria were positively correlated with HAMD.The Actinobacteria were positively correlated with UPDRS-III.3.TNF-α was positively correlated with Firmicutes,actinobacteria and Clostridia.There was a negative correlation with Bacteroidetes.There was no correlation between IL-8 and the relative abundance of intestinal flora in PD.4.Roseburia may be related to liver wind internal motion syndrome;the turbidities of phlegm may be related to the abundance of microflora in Verrucomicrobia,Verrucomicrobiae,Verrucomicrobiales,and Verrucomicrobiaceae.