Professor Ding E's Oral Administration Of Huayu Tongluo Method And Fumigation And Washing On The Clinical Efficacy Of Knee Osteoarthritis,the Effect Of IL-17 And IL-37 Expression Level And Its Mechanism

1 PurposeTo observe the changes of TNF-?,IL-10,IL-17,IL-37,VEGF,Treg and other laboratory indicators,VAS score,Lequesne score,WOMAC score,TCM syndrome score and symptom quantification score and other indicators in knee osteoarthritis patients?KOA?,to explore the mechanism and correlation analysis of the effects of oral administration and fumigation of Huayu Tongluo method on TNF-?,IL-10,IL-17,IL-37,VEGF and Treg and other indicators in KOA patients,and to study its clinical efficacy on KOA patients.2 Methods2.1 Theoretical ResearchThrough previous literature research,the correlation between TCM"blood stasis"and laboratory indexes of KOA patients such as TNF-?,IL-10,IL-17,IL-37,VEGF and Treg was analyzed,and the close relationship between TCM etiology and pathogenesis of KOA patients and"blood stasis"was analyzed,which provided theoretical basis for oral administration and fumigation treatment of KOA with Huayu Tongluo method.2.2 Clinical Research2.2.1 Changes and Correlation of TNF-?,IL-10,IL-17,IL-37 and Related Indicators in KOA Patients20 health personnel from Anhui traditional Chinese medicine hospital health management center were selected as normal control group,and 60 KOA patients from Anhui traditional Chinese medicine hospital rheumatology department in-patient and out-patient were selected as KOA group.there was no significant difference in general data between the two groups?p>0.05?.The VAS score,Lequesne score,WOMAC score,TCM syndrome score and symptom quantification score of 60 KOA patients were observed.ELLSA was used to detect TNF-?,IL-10,IL-17,IL-37 in normal control group and patients in KOA group.PCR was used to detect m RNA of IL-17,IL-37.Meanwhile,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg,PLT,D-D,ESR,hs-CRP and other laboratory indicators were detected.SPSS 22.0 was used to analyze the above data.2.2.2 Effect of Oral Administration and Fumigation of Huayu Tongluo Method on Clinical Efficacy,Quality of Life and Related Experimental Indicators of KOA Patients60 KOA patients were selected from the department of rheumatology of Anhui traditional Chinese medicine hospital,which met the inclusion criteria.60 patients were divided into the control group and the research group with 30 cases in each group according to the random number table method.Oral glucosamine sulfate capsules were used as the basic drugs for the two groups.The control group was treated by oral administration of traditional Chinese medicine and the research group was treated by oral administration of traditional Chinese medicine plus fumigation and washing.The changes of laboratory indexes such as TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg,PLT,D-D,ESR,hs-CRP,VAS score,Lequesne score,WOMAC score,TCM syndrome score,symptom quantification score and quality of life were observed.SPSS 22.0 was used to analyze the above data.3 Results3.1 Theoretical Analysis Results?1?The occurrence and development of KOA are closely related to TNF-?,IL-10,IL-17,IL-37,Treg,VEGF and other indicators.?2?"Blood Stasis"runs through KOA from beginning to end;?3?TNF-?,IL-10,IL-17,IL-37,Treg,VEGF are closely related to"blood stasis".?4?Huayu Tongluo method is an important treatment for KOA.3.2 Clinical Research Results3.2.1 Comparison of TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg between KOA group and normal control groupCompared with the normal control group,the levels of TNF-?,IL-17 and VEGF in KOA group increased?P<0.01?,while the levels of IL-10,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg decreased?P<0.01?.3.2.2 Comparison of m RNA of IL-17 and IL-37 in Peripheral Blood Mononuclear Cells between KOA Group and Normal Control GroupCompared with the normal control group,the m RNA level of IL-17 in peripheral blood mononuclear cells of patients in KOA group increased?P<0.01?and the m RNA level of IL-37 decreased?P<0.01?.3.2.3 Comparison of Laboratory Indexes between KOA Group and Normal Control GroupCompared with the normal control group,CRP,ESR,D-D,FIB,TT,PLT,PCT in KOA patients increased?P<0.05 or P<0.01?,while APTT,PT,PDW,MPV decreased?P<0.01?.3.2.4 Correlation Study of TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg and Other Indicators in KOA Group?1?The correlation between TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg and symptom grading quantitative score and TCM syndrome score in KOA groupThe results showed that TNF-?in KOA patients was positively correlated with symptom grading quantitative score,joint pain and joint swelling?P<0.05 or P<0.01?.IL-10 was negatively correlated with joint swelling?P<0.05?.IL-17 was positively correlated with symptom grading quantitative score and joint swelling?P<0.05?.IL-37was negatively correlated with symptom grading quantitative score,joint pain and joint swelling?P<0.05 or P<0.01?.VEGF was positively correlated with symptom grading quantitative score and joint swelling?P<0.05?.?2?The correlation between TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg and VAS score,Lequesne score,WOMAC score,SF-36score in KOA groupThe results showed that TNF-?in KOA patients was positively correlated with VAS score,Lequesne score and WOMAC score?P<0.01?,but negatively correlated with physiological function,physiological function,social function,physical pain and mental health?P<0.05 or P<0.01?.IL-10 was negatively correlated with VAS score,Lequesne score and WOMAC score?P<0.05 or P<0.01?,and was positively correlated with physiological function,physical pain and mental health?P<0.01?.IL-17 was positively correlated with VAS score,Lequesne score and WOMAC score?P<0.05 or P<0.01?,but negatively correlated with physiological function,physical pain and mental health?P<0.05 or P<0.01?.IL-37 was negatively correlated with VAS score,Lequesne score and WOMAC score?P<0.01?,and was positively correlated with mental health?P<0.01?.VEGF was positively correlated with Lequesne score and WOMAC score?P<0.05 or P<0.01?,but negatively correlated with physiological function?P<0.01?.CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg was negatively correlated with Lequesne score?P<0.01?.?3?The correlation between TNF-?,IL-10,IL-17,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg and laboratory indexes in KOA groupThe results showed that TNF-?was positively correlated with CRP,ESR and PLT in KOA patients?P<0.01?.IL-10 was negatively correlated with ESR and PLT?P<0.05 or P<0.01?.IL-17 was positively correlated with CRP,ESR,D-D,PLT?P<0.05 or P<0.01?.IL-37 was negatively correlated with ESR and PLT?P<0.05 or P<0.01?.VEGF was positively correlated with CRP,FIB and PCT?P<0.05 or P<0.01?.CD4⁺CD25⁺CD127 ^low/-Treg was negatively correlated with FIB,PT and PLT?P<0.05or P<0.01?.?4?The correlation between TNF-?,IL-10,IL-17,IL-37 and VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg in KOA groupThe results showed that TNF-?in KOA patients was positively correlated with VEGF?P<0.01?and negatively correlated with CD4⁺CD25⁺Treg?P<0.01?.IL-10 was positively correlated with CD4⁺CD25⁺Treg?P<0.01?.IL-17 was positively correlated with VEGF?P<0.05?.IL-37 was negatively correlated with VEGF?P<0.05?,and positively correlated with CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg?P<0.05 or P<0.01?.3.2.5 Clinical Efficacy of Oral Administration of Huayu Tongluo Method and Fumigation for KOA PatientsCompared with the control group,there was no significant difference in clinical therapeutic effect between the two groups?P>0.05?.However,the significant efficiency of the study group was higher than that of the control group?P<0.05?.3.2.6 Effects of Oral Administration of Huayu Tongluo Method and Fumigation on TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-TregAfter treatment,TNF-?,IL-17 and VEGF in both groups decreased significantly?P<0.01?,while IL-10,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg in both groups increased significantly?P<0.01?.After treatment,TNF-?,IL-17 and VEGF in the study group decreased significantly?P<0.05 or P<0.01?,while IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127^low/-Treg increased significantly?P<0.05 or P<0.01?.3.2.7 Comparison of m RNA of IL-17 and IL-37 in Peripheral Blood Mononuclear Cells by Oral Administration and Fumigation of Huayu Tongluo MethodAfter treatment,the m RNA of IL-17 in both groups decreased significantly?P<0.01?,while the m RNA of IL-37 in both groups increased significantly?P<0.01?.After treatment,the m RNA level of IL-37 in the study group increased significantly?P<0.01?.3.2.8 Effect of Oral Administration and Fumigation of Huayu Tongluo Method on Laboratory IndexesAfter treatment,CRP,ESR,D-D,FIB,PLT and PCT in both groups decreased?P<0.01or P<0.05?,APTT,PT and MVP increased?P<0.01?.After treatment,the improvement of CRP,D-D and PCT in the study group was more obvious than that in the control group?P<0.05?.3.2.9 Effect of Oral Administration and Fumigation of Huayu Tongluo Method on TCM Syndrome Score,Symptom Quantification Score,VAS Score,Lequesne Score and WOMAC ScoreAfter treatment,the two groups played a role in reducing the total score of TCM syndromes,quantitative score of symptoms,VAS score,Lequesne score and WOMAC score?P<0.01?,and the research group was superior to the control group in reducing the score of TCM syndromes,quantitative score of symptoms,Lequesne score and WOMAC score?P<0.05 or P<0.01?.3.2.10 Effect of Oral Administration and Fumigation of Huayu Tongluo Method on Scores of SF-36 Dimensions of Quality of Life ScaleAfter treatment,the study group's overall health,physiological function,physiological function,emotional function,social function,physical pain and energy dimension scores increased significantly?P<0.01 or P<0.05?,while the control group's overall health and physical pain dimension scores increased significantly?P<0.01 or P<0.05?.Compared with the two groups after treatment,the improvement of physiological function and physical pain dimension scores in the study group were better than those in the control group?P<0.05?.4 Conclusion4.1 The disease progression of KOA is related to the levels of TNF-?,IL-10,IL-17,IL-37,Treg and VEGF.4.2 The occurrence and development of KOA are closely related to"blood stasis"."Huayu Tongluo Method"is an important therapeutic method for KOA.4.3 The levels of TNF-?,IL-17,VEGF,IL-17m RNA in KOA patients increased,while the levels of IL-10,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg,IL-37m RNA decreased.4.4 TNF-?,IL-10,IL-17,IL-37,VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg,in KOA patients are correlated with TCM syndrome score,VAS score,Lequesne score,WOMAC score and related laboratory indexes.4.5 TNF-?,IL-10,IL-17,IL-37 in KOA patients are correlated with VEGF,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg;4.6 Oral administration and fumigation with Huayu Tongluo Method have significant clinical effects,obviously improving the joint symptoms of patients and improving the quality of life;4.7 Oral administration and fumigation with Huayu Tongluo Method can obviously reduce the levels of TNF-?,IL-17,VEGF and IL-17m RNA in patients,increase the levels of IL-10,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127 ^low/-Treg,IL-37m RNA in patients,and obviously improve the coagulation function and platelet related indexes.4.8 Possible Intervention Mechanism of Professor Ding E's Huayu Tongluo Method for Oral Administration and Fumigation on KOAProfessor Ding E's Huayu Tongluo Method for oral administration and fumigation may protect knee osteoarthritis by lowering IL-17m RNA level,increasing IL-37m RNA level,lowering TNF-?,IL-17 and VEGF level,increasing IL-10,IL-37,CD4⁺CD25⁺Treg,CD4⁺CD25⁺CD127^low/-Treg,level,inhibiting pro-inflammatory reaction,increasing anti-inflammatory reaction,improving coagulation function and platelet related indexes,relieving hypercoagulable state.