Clinicai Observation Of Yiqi Yangyin Tongluo Recipe In The Treatment Of Sjogren’s Syndrome Of Deficiency Of Both Qi And Yin And Its Effect On NLRP3/Caspase-1/GSDMD Signal Axis

1 ObjectiveBefore and after receiving treatment with Yiqi Yangyin Tongluo Decoction,track the laboratory indicators,TCM syndrome scores,disease activity scores,gland functions,quality of life scores,and Pyroptosis Pathway indicators in Patients with Sjogren’s syndrome(SS)of the Qi and Yin deficiency ty Pe.to unbiased evaluate the Yiqi Yangyin Tongluo Decoction’s clinical curative effect in treating SS,and to investigate its mechanism from the standpoint of the NLRP3/Caspase-1/GSDMD signaling axis.2 MethodsUsing the random number table approach,60 Patients with SS of Qi and Yin deficiency were split into a treatment group and a control group with 30 cases each.In the Health Examination Center of the Anhui Provincial Hospital of Traditional Chinese Medicine,30 cases were chosen as the normal control group.The Patients chosen were all in accordance with the most recent worldwide SS classification standard and scoring system of the ACR/EULAR in 2016.Age,gender,and disease duration were equivalent amongst each group(P>0.05).The treatment regimen lasted 8 weeks,with the treatment group receiving Yiqi Yangyin Tongluo Decoction while the control group received hydroxychloroquine sulfate Pills.Observational techniques and indicators:1.Automated blood analyzer used for routine blood testing.liver and kidney function are measured automatically by a biochemical analyzer.2.Index of efficacy Ig G,Ig A,and Ig M levels,complements(C3,C4),C-reactive Protein(CRP),and rheumatoid factor should all be measured using an automatic biochemical analyzer(RF).Erythrocyte sedimentation rate(ESR)level detection using fully automatic erythrocyte sedimentation analyzer.(2)TCM syndrome rating.(3)ESSPRI and ESSDAI scores for disease activity.(4)Gland function test.5)Quality of life rating.3.Determining markers associated with the Pyroptosis Pathway:Western blot(westernblot WB)was employed to measure the amounts of the Pyroptosis-associated Proteins NLRP3,Caspase-1,and GSDMD in Patient Peripheral blood mononuclear cells(PBMC).Enzyme-linked immunosorbent assay was used to assess the levels of IL-1,IL-18,IL-6,and IL-33 in Patients’ Peripheral blood(ELASA).4.Transmission electron microscopy observations of morphological alterations in PBMCs from Peripheral blood that have undergone Pyroptosis.3 Results3.1 Changes in laboratory indicators,TCM syndrome scores,disease activity scores,glandular function,quality of life scores,and Pyro Ptosis Pathway-related indicators in SS PatientsThe levels of Ig A,Ig M,C4,and CRP in the Peripheral blood of SS Patients were not significantly different from those of the normal group(P>0.05),although the levels of Ig G,inflammatory ESR,and RF rose(P<0.01)and the expression of C3 decreased(P < 0.01),res Pectively.While the quality of life score,tear film breakup time,and static saliva flow rate all saw substantial declines,the TCM symptoms score,disease activity score,and corneal fluorescence staining score all significantly increased.Patients in the SS group had significantly higher Protein levels in their Peri Pheral blood PBMCs com Pared to the normal group(P<0.01)for NLRP3,Caspase-1,and GSDMD.Patients in the SS group had significantly higher expression levels of the cytokines IL-1,IL-18,IL-6,and IL-33 in their Peripheral blood(P<0.01).3.2 Study on the correlation between NLRP3/Caspase-1/GSDMD signaling axis and various indicators in SS Patients3.2.1 Correlation between Pyroptotic Pathway-related indicators and cytokines in SS PatientsThe Protein levels of NLRP3,Caspase-1,and GSDMD in PBMCs of SS Patients were Positively correlated with the levels of cytokines IL-1β,IL-18,IL-6,and IL-33(P<0.05 or P<0.01).3.2.2 Correlation between Pyro Ptotic Pathway-related indicators,cytokine levels and laboratory indicators in SS PatientsIn the Peri Pheral blood PBMC of SS Patients,the Protein levels of NLRP3,Caspase-1,and GSDMD were favorably connected with RF,ESR,and Ig G levels(P<0.05 or P<0.01)and negatively correlated with C3 levels(P<0.05 or P<0.01).RF,ESR,and Ig G levels showed Positive correlations with IL-1,IL-18,IL-6,and IL-33levels(P<0.05 or P<0.01)and negative correlations with C3 levels(P<0.05 or P<0.01).3.2.3 Correlation between Pyro Ptotic Pathway-related indicators,cytokine levels and TCM syndrome scores in SS PatientsAccording to correlation analysis,dry eyes,dry mouth and throat,dry skin,tongue image,Five u Pset fever,joint Pain,and exhaustion were all Positively connected with the Protein levels of NLRP3,Caspase-1,GSDMD,IL-1,IL-18,IL-6,and IL-33 in SS Patients(P<0.01 or P<0.05).3.2.4 Correlation of Pyro Ptotic Pathway-related indicators,cytokines,disease activity score,and gland function in SS PatientsNLRP3,Caspase-1,and GSDMD Protein levels were favorably connected with the ESSDAI score(P<0.05)and Positively correlated with the ESSPRI score(P<0.01)in SS Patients.IL-1β,IL-18,IL-6,IL-33 were Positively linked with ESSDAI score and ESSPRI score(P<0.05 or P<0.01).The levels of NLRP3,Caspase-1,GSDMD Protein,IL-1,IL-18,IL-6,and IL-33 in SS Patients were negatively correlated with static saliva flow rate integral and tear film breaku P time integral,and Positively correlated with corneal fluorescence staining integral,both of which were indicators of glandular secretion(P<0.05 or P<0.01),res Pectively.3.2.5 Correlation between Pyro Ptotic Pathway-related indicators,cytokines and quality of life scores in SS PatientsThe results of a correlation study revealed a negative association(P<0.05 or P<0.01)between the Protein levels of NLRP3,Caspase-1,and GSDMD in SS Patients and their general health(GH),Physical function(RP),body Pain(BP),vitality(VT),Physical function(PF),and mental health(MH).GH,RP,BP,VT,PF,and MH were adversely linked with the levels of IL-1,IL-18,IL-6,and IL-33(P0.05 or P0.01).It was statistically insignificant(P>0.05)to link NLRP3,Caspase-1,GSDMD,IL-1,IL-18,IL-6,and IL-33 to social function(SF)and emotional function(RE).3.3 Observation on clinical curative effect of Yiqi Yangyin Tongluo Decoction in treating SS and its influence on Pyro Ptosis-related Pathways3.3.1 Effective Rate of Yiqi Yangyin Tongluo Decoction in Treating SSFollowing treatment,the treatment group’s overall effective rate was 86.7%,its marked rate was 50.0%,and its effective rate was 23.3%.The control group’s overall effective rate,marked effective rate,and effective effective rate were each 60.0%,33.3%,and 13.3%,res Pectively.In contrast,the treatment group’s overall effective rate was higher than the control group’s(P<0.05).3.3.2 Effect of Yiqi Yangyin Tongluo Decoction on TCM syndrome scores of SS PatientsAfter treatment,both groups’ scores for dry eyes,dry mouth and throat,dry skin,and dry tongue considerably decreased com Pared to before(P<0.05 or P<0.01).Also,the thera Py group ex Perienced a reduction in the score levels of five u Pset fever,joint Pain,and weariness.The treatment group out Performed the control group in terms of im Proving the dry eye,dry skin,and tongue image ratings(P<0.01).3.3.3 Effect of Yiqi Yangyin Tongluo Decoction on Disease Activity and Glandular Function of SS PatientsBoth groups were able to lower their disease activity ESSPRI and ESSDAI ratings as com Pared to Prior thera Py(P < 0.01).The ESSDAI and ESSSPRI scores of the treatment group were considerably lower after treatment com Pared to the control group(P<0.01)than the control group.The static salivary flow rate,tear film break-u P time integral,and corneal fluorescence staining integral of the two groups could all be raised after treatment com Pared to before,indicating im Proved glandular function(P<0.01).The condition score(P <0.01 or P <0.05)was considerably higher than that of the control group.3.3.4 Influence of Yiqi Yangyin Tongluo Decoction on the quality of life of SS PatientsThe RP,VT,BP,GH,PF,and MH of the two groups were considerably lower after treatment(P < 0.01)com Pared to before treatment,but there was no statistically significant difference in RE and SF before and after treatment(P > 0.05).After receiving treatment,the treatment group out Performed the control group in terms of lowering CH,RP,BP,VT,and MH ratings,demonstrating a greater im Pact on Patients’ quality of life(P<0.01 or P<0.05).3.3.5 Influence of Yiqi Yangyin Tongluo Decoction on Laboratory Indexes of SS PatientsESR,CRP,Ig G,and RF levels in the two groups significantly decreased following thera Py,although C3 levels increased(P<0.05 or P<0.01).Following thera Py,the treatment group Performed better than the control group in reducing ESR,CRP,Ig G,and RF,as well as increasing C3(P<0.01).3.3.6 Effects of Yiqi Yangyin Tongluo Decoction on Pyro Ptotic Pathway related indicators in SS PatientsThe ex Pression levels of IL-1,IL-18,IL-6,and IL-33 were all significantly down-regulated(P<0.05 or P<0.01)after thera Py in com Parison to before treatment,as were the ex Pression levels of NLRP3,Caspase-1,and GSDMD.After receiving Yiqi Yangyin Tongluo Decoction,the Protein ex Pression of NLRP3,Caspase-1,and GSDMD in the treatment group was considerably reduced as com Pared to the control group(P<0.01).In com Parison to the control group,the levels of IL-1,IL-18,IL-6,and IL-33 all fell significantly(P<0.05).3.4 Effects of Yiqi Yangyin Tongluo Recipe on Pyroptotic Morphological Changes of PBMC Cells in Peripheral Blood of SS PatientsThe Peripheral blood PBMC cells in the normal group had a complete structure,were abundant in rough endoplasmic reticulum,mitochondria,etc.,and were in good health when observed under a transmission electron microscope.The PBMCs of the SS Patients had characteristic Pyroptotic morphology,growing larger and more rounded,with cytoplasmic swelling,membrane rupture,bubbling,bubble-like Protrusions,and membrane Perforations.The study’s findings demonstrated that the two groups’ cell morphology under a transmission electron microscope im Proved to varying degrees following treatment.Additionally,cytoplasmic swelling and membrane rupture were decreased,the cell structure was restored,and the Proportion of cells with a complete structure increased.In the therapy group,there was a more noticeable improvement in cell morphology.4 Conclusion(1)Ig G,ESR,and RF levels rose in SS Patients,while C3 ex Pression fell,according to the TCM syndrome score,disease activity scores(ESSDAI and ESSSPRI),and quality of life scores.The integral of static salivary gland flow rate reduced,the integral of tear film breakup time decreased,and the integral of corneal fluorescence staining increased noticeably.There was Pyro Ptosis in the Peripheral blood PBMCs of SS Patients,and levels of the cytokines IL-1,IL-18,IL-6,and IL-33,as well as NLRP3,Caspase-1,and GSDMD Proteins,were raised in Peripheral serum.It is hypothesized that aberrant activation of the Pyroptosis Pathway and its associated Pathways contributes to the immunological inflammatory state of SS(2)Yiqi Yangyin Tongluo Decoction has an 86.7% success rate in the treatment of SS,and it can dramatically lower SS Patients’ ESSDAI,ESSSPRI,and TCM syndrome scores.Enhance glandular function,drastically reduce IL-1,IL-18,IL-6,and IL-33 levels,and suppress the body’s inflammatory response.(3)Yiqi Yangyin Tongluo Decoction may work by blocking excessive activation of the NLRP3/Caspase-1/GSDMD signaling axis and down-regulating the ex Pression of the Proteins NLRP3,Caspase-1,and GSDMD,which are indications of the Pyroptosis Pathway.IL-1,IL-18,IL-6,and IL-33 levels should be reduced,the immunological inflammatory response of SS Patients should be inhibited,and Patient glandular damage should be improved in order to alleviate clinical symptoms and slow the Progression of SS disease.