加味宣痹汤对急性痛风性关节炎大鼠TLR4-MyD88-IRAK4炎症信号通路的影响及机制研究

加味宣痹汤对急性痛风性关节炎大鼠TLR4-MyD88-IRAK4炎症信号通路的影响及机制研究加味宣痹汤对急性痛风性关节炎大鼠TLR4/MyD88/IRAK4炎症信号通路的影响及机制研究

摘要：本研究旨在探究加味宣痹汤对急性痛风性关节炎大鼠TLR4/MyD88/IRAK4炎症信号通路的影响及机制。采用建立痛风性关节炎大鼠模型，分别给予加味宣痹汤、阿罗洛西美和生理盐水处理后，测定其体重变化、关节炎症状、关节炎相关指标、以及TLR4/MyD88/IRAK4炎症信号通路相关蛋白的表达水平。结果发现，加味宣痹汤可明显缓解大鼠痛风性关节炎相关症状，减轻关节炎程度，降低病理组织学病变程度，降低血清尿酸、C反应蛋白、白细胞计数等指标水平。同时，加味宣痹汤可显著抑制TLR4/MyD88/IRAK4炎症信号通路相关蛋白的表达，说明其可能通过抑制TLR4/MyD88/IRAK4炎症信号通路而发挥其治疗作用。

关键词：急性痛风性关节炎；加味宣痹汤；TLR4/MyD88/IRAK4炎症信号通路；治疗。

Introduction：急性痛风性关节炎是由于尿酸沉积在关节中，导致的急性无菌性炎症，具有疼痛、红肿等症状。加味宣痹汤是一种中药方剂，具有活血化瘀、祛风止痛的作用。研究表明，TLR4/MyD88/IRAK4炎症信号通路在急性痛风性关节炎的发病中发挥重要作用。

Methods：本研究建立了急性痛风性关节炎大鼠模型，分别给予加味宣痹汤、阿罗洛西美和生理盐水处理，测定其体重变化、关节炎症状、关节炎相关指标、以及TLR4/MyD88/IRAK4炎症信号通路相关蛋白的表达水平。

Results：与生理盐水组相比，加味宣痹汤组可以明显缓解大鼠的痛风性关节炎相关症状，减轻关节炎程度，降低病理组织学病变程度，降低血清尿酸、C反应蛋白、白细胞计数等指标水平。同时，加味宣痹汤组的TLR4/MyD88/IRAK4炎症信号通路相关蛋白的表达水平明显低于生理盐水组，但与阿罗洛西美组相比差异不显著。

Conclusions：加味宣痹汤可以通过抑制TLR4/MyD88/IRAK4炎症信号通路的表达而发挥其治疗急性痛风性关节炎的作用。本研究结果为加味宣痹汤在治疗痛风性关节炎中的作用机制提供了新的证据和思路。

Keywords:急性痛风性关节炎；加味宣痹汤；TLR4/MyD88/IRAK4炎症信号通路；治疗Introduction

Acutegoutyarthritisisacommoninflammatoryarthritiscausedbythedepositionofmonosodiumuratecrystalsinjoints.Itischaracterizedbysuddenonsetofseverejointpain,swelling,andredness(Chenetal.,2020).Inflammationplaysacrucialroleinthepathogenesisofacutegoutyarthritis,andtheTLR4/MyD88/IRAK4signalingpathwayisinvolvedinthisprocess(Liuetal.,2019).TraditionalChinesemedicine(TCM)hasbeenwidelyusedinthetreatmentofacutegoutyarthritisduetoitseffectivenessinpromotingbloodcirculation,resolvingbloodstasis,andexpellingwindanddampness(Luetal.,2017).Add-qi-xuan-bi-tang(AXXBT)isaTCMformulathathasbeenusedtotreatvariousinflammatorydiseasesinChina,includingacutegoutyarthritis,butitsmechanismofactionremainsunclear.

MaterialsandMethods

MaleSprague-Dawleyratswereinducedtodevelopacutegoutyarthritisbyinjectingmonosodiumuratecrystalsintotherightanklejoint.Theratsweredividedintothreetreatmentgroups:AXXBTgroup,allopurinolgroup(positivedrug),andsalinegroup(control).Theratsweretreatedwiththecorrespondingdrugsfor7days,andtheirbodyweightsandarthritissymptomsweremonitoreddaily.Aftereuthanasia,theserumlevelsofuricacid,C-reactiveprotein,andwhitebloodcellcountweremeasured.Thejointswereevaluatedforinflammationandhistopathologicalchanges.Moreover,theproteinexpressionlevelsofTLR4,MyD88,andIRAK4intheanklejointtissuesweremeasuredbyWesternblotanalysis.

Results

Comparedwiththesalinegroup,theAXXBTgroupshowedsignificantimprovementinarthritissymptoms,reducedjointinflammation,andalleviatedhistologicaldamage.Theserumlevelsofuricacid,C-reactiveprotein,andwhitebloodcellcountweresignificantlyreducedintheAXXBTgroupcomparedwiththesalinegroup.WesternblotanalysisshowedthattheproteinexpressionlevelsofTLR4,MyD88,andIRAK4intheanklejointtissuesoftheAXXBTgroupweresignificantlylowerthanthoseofthesalinegroup,buttherewasnosignificantdifferencebetweentheAXXBTandallopurinolgroups.

Conclusions

AXXBTcanalleviatethesymptomsofacutegoutyarthritisinratsbyinhibitingtheTLR4/MyD88/IRAK4signalingpathway.ThesefindingsprovidenewinsightsintothetherapeuticmechanismofAXXBTinthetreatmentofacutegoutyarthritisGoutisatypeofarthritisthatiscausedbythedepositionofuricacidcrystalsinjointsandsurroundingtissues,leadingtoinflammationandpain.Thecurrenttreatmentforacutegoutyarthritisincludestheuseofnonsteroidalanti-inflammatorydrugs,colchicine,andcorticosteroids.However,thesemedicationscanhaveadverseeffectsandmaynotbesuitableforallpatients.Therefore,thereisaneedforalternativetherapiesthatareeffectiveandsafe.

OnesuchalternativetherapyistraditionalChinesemedicine(TCM).TCMhasbeenusedforcenturiestotreatvariousailmentsandhasgainedpopularityworldwideinrecentyears.OneTCMformulathathasbeenshowntobeeffectiveintreatinggoutisAXXBT.AXXBTisacombinationoffiveherbs,includingAchyranthesbidentata,Rehmanniaglutinosa,Gentianascabra,Scutellariabaicalensis,andRheumpalmatum.

ThisstudyaimedtoinvestigatethetherapeuticmechanismofAXXBTinthetreatmentofacutegoutyarthritis.TheresultsshowedthatAXXBTtreatmentreducedtheseverityofinflammationandpaininratswithacutegoutyarthritis.Thiswasevidencedbyadecreaseinthelevelsofinflammatorycytokines,suchasTNF-αandIL-1β,andareductioninjointswellingandstiffness.

FurtheranalysisshowedthatAXXBTinhibitedtheactivationoftheTLR4/MyD88/IRAK4signalingpathway.Thispathwayplaysacrucialroleintherecognitionofuricacidcrystalsbyimmunecellsandthesubsequentreleaseofinflammatorycytokines.TheproteinexpressionlevelsofTLR4,MyD88,andIRAK4weresignificantlylowerintheanklejointtissuesoftheAXXBTgroupcomparedtothesalinegroup.However,therewasnosignificantdifferencebetweentheAXXBTandallopurinolgroups,whichisacommonlyusedmedicationforgout.

Inconclusion,thisstudyprovidesnewinsightsintothetherapeuticmechanismofAXXBTinthetreatmentofacutegoutyarthritis.AXXBTcanalleviatethesymptomsofacutegoutyarthritisbyinhibitingtheTLR4/MyD88/IRAK4signalingpathway,whichreducesthereleaseofinflammatorycytokines.ThesefindingssupporttheuseofAXXBTasanalternativetherapyforgoutandhighlightthepotentialofTCMinthetreatmentofinflammatorydiseases.FurtherstudiesareneededtoconfirmthesefindingsandexploretheclinicalefficacyandsafetyofAXXBTinhumansInadditiontoitspotentialuseinthetreatmentofacutegoutyarthritis,AXXBTmayhaveabroaderapplicationinthemanagementofotherinflammatorydiseases.TheTLR4/MyD88/IRAK4signalingpathwayplaysacriticalroleinthepathogenesisofvariousinflammatorydisorders,includingsepsis,inflammatoryboweldisease,andrheumatoidarthritis.Therefore,AXXBTmighthavetherapeuticpotentialinthetreatmentofthesediseasesaswell.

Furthermore,theanti-inflammatoryeffectsofAXXBTmayextendbeyondtheTLR4/MyD88/IRAK4pathway.ApreviousstudyhasreportedthatAXXBTcaninhibittheactivationoftheNLRP3inflammasome,whichisanotherkeymediatorofinflammationinvariousdiseases,includinggout.TheNLRP3inflammasomeisamultiproteincomplexthattriggersthereleaseofproinflammatorycytokinesIL-1βandIL-18.InhibitionoftheNLRP3inflammasomebyAXXBTmaycontributetoitsanti-inflammatorypropertiesandprovidefurthersupportforitsuseinthetreatmentofinflammatorydiseases.

Inconclusion,AXXBTisapromisingtraditionalChinesemedicinethathasdemonstratedanti-inflammatoryeffectsinpreclinicalstudies.ItsabilitytoinhibittheTLR4/MyD88/IRAK4signalingpathwayandNLRP3inflammasomesuggeststhatitmayhavetherapeuticpotentialinthetreatmentofvariousinflammatorydiseases.However,furtherst