苓桂術(shù)甘湯含藥血清通過PI3KAkt信號(hào)通路保護(hù)H2O2誘導(dǎo)的H9c2細(xì)胞損傷

苓桂術(shù)甘湯含藥血清通過PI3KAkt信號(hào)通路保護(hù)H2O2誘導(dǎo)的H9c2細(xì)胞損傷一、本文概述Overviewofthisarticle本文旨在探討苓桂術(shù)甘湯含藥血清對(duì)H2O2誘導(dǎo)的H9c2細(xì)胞損傷的保護(hù)作用及其機(jī)制。通過深入研究PI3K/Akt信號(hào)通路在此過程中的作用，我們期望為心血管疾病的治療提供新的藥物候選和理論依據(jù)。實(shí)驗(yàn)將采用H9c2細(xì)胞作為心肌細(xì)胞的代表，模擬氧化應(yīng)激損傷環(huán)境，觀察苓桂術(shù)甘湯含藥血清對(duì)細(xì)胞存活率、凋亡及相關(guān)蛋白表達(dá)的影響。我們將分析PI3K/Akt信號(hào)通路的激活情況，以揭示苓桂術(shù)甘湯含藥血清保護(hù)心肌細(xì)胞的分子機(jī)制。這一研究不僅有助于深入理解傳統(tǒng)中藥苓桂術(shù)甘湯的藥理作用，還可能為開發(fā)新型心血管疾病治療藥物提供重要線索。ThisarticleaimstoexploretheprotectiveeffectandmechanismofLingguiZhuganTangcontainingserumonH2O2induceddamagetoH9c2cells.Byconductingin-depthresearchontheroleofthePI3K/Aktsignalingpathwayinthisprocess,wehopetoprovidenewdrugcandidatesandtheoreticalbasisforthetreatmentofcardiovasculardiseases.TheexperimentwilluseH9c2cellsasrepresentativesofmyocardialcells,simulateoxidativestressdamageenvironment,andobservetheeffectsofLingguiZhuganTangcontainingserumoncellsurvivalrate,apoptosis,andrelatedproteinexpression.WewillanalyzetheactivationofthePI3K/AktsignalingpathwaytorevealthemolecularmechanismofLingguiZhuganTangcontainingseruminprotectingmyocardialcells.ThisstudynotonlycontributestoadeeperunderstandingofthepharmacologicaleffectsoftraditionalChinesemedicineLingguiZhuganTang,butmayalsoprovideimportantcluesforthedevelopmentofnewdrugsforthetreatmentofcardiovasculardiseases.二、材料與方法MaterialsandMethods苓桂術(shù)甘湯：購自正規(guī)中藥房，經(jīng)嚴(yán)格鑒定確保其質(zhì)量。H2O2（過氧化氫）：購自Sigma-Aldrich公司。胎牛血清（FBS）：購自HyClone公司。DMEM高糖培養(yǎng)基、胰蛋白酶：購自Gibco公司。PI3K抑制劑LY294002：購自CellSignalingTechnology公司。兔抗Akt、兔抗p-Akt抗體：購自Abcam公司。HRP標(biāo)記的山羊抗兔IgG：購自北京中杉金橋生物技術(shù)有限公司。LingGuiZhuGanTang:purchasedfromalegitimateChinesepharmacy,anditsqualityisensuredthroughstrictidentification.H2O2(hydrogenperoxide):purchasedfromSigmaAldrichcompany.Fetalbovineserum(FBS):purchasedfromHyClonecompany.DMEMhighsugarculturemediumandtrypsin:purchasedfromGibcocompany.PI3KinhibitorLY294002:purchasedfromCellSignalingTechnology.RabbitantiAktandrabbitantip-Aktantibodies:purchasedfromAbcamcompany.HRPlabeledgoatantirabbitIgG:purchasedfromBeijingZhongshanJinqiaoBiotechnologyCo.,Ltd.細(xì)胞培養(yǎng)箱：購自ThermoFisherScientific公司。倒置顯微鏡：購自O(shè)lympus公司。酶標(biāo)儀：購自Bio-Rad公司。WesternBlot電泳及轉(zhuǎn)膜設(shè)備：購自Bio-Rad公司。Cellcultureincubator:purchasedfromThermoFisherScientific.Invertedmicroscope:purchasedfromOlympuscompany.Enzymereader:purchasedfromBioRadcompany.WesternBlotelectrophoresisandmembranetransferequipment:purchasedfromBioRadcompany.將苓桂術(shù)甘湯按成人臨床等效劑量（根據(jù)人與動(dòng)物的體表面積換算）給健康SD大鼠灌胃，連續(xù)灌胃7天后，無菌采集大鼠靜脈血，離心分離血清，經(jīng)56℃滅活30分鐘，即得苓桂術(shù)甘湯含藥血清。LingGuiZhuGanTangwasorallyadministeredtohealthySDratsatanequivalentadultclinicaldose(convertedbasedonthebodysurfaceareaofhumansandanimals).After7consecutivedaysofgastricadministration,sterilevenousbloodwascollectedfromtherats,andtheserumwascentrifugedandseparated.Afterbeinginactivatedat56℃for30minutes,thedrugcontainingserumofLingGuiZhuGanTangwasobtained.H9c2細(xì)胞用含10%FBS的DMEM高糖培養(yǎng)基，在37℃、5%CO2的細(xì)胞培養(yǎng)箱中培養(yǎng)。待細(xì)胞長(zhǎng)至80%融合時(shí)，用25%胰蛋白酶消化傳代。H9c2cellswereculturedinDMEMhighglucosemediumcontaining10%FBSinacellcultureincubatorat37℃and5%COWhenthecellsgrowto80%fusion,theyaredigestedandpassagedwith25%trypsin.將H9c2細(xì)胞接種于96孔板或6孔板中，待細(xì)胞貼壁后，用不同濃度的H2O2處理細(xì)胞，通過CCK-8法檢測(cè)細(xì)胞活力，確定最佳造模濃度。InoculateH9c2cellsintoa96wellplateora6-wellplate,andafterthecellsadheretothewall,treatthecellswithdifferentconcentrationsofH2ODetectcellviabilityusingtheCCK-8methodtodeterminetheoptimalmodelingconcentration.實(shí)驗(yàn)分為正常對(duì)照組、模型組、苓桂術(shù)甘湯含藥血清組、PI3K抑制劑組及聯(lián)合用藥組。除正常對(duì)照組外，其余各組均用最佳造模濃度的H2O2處理細(xì)胞。苓桂術(shù)甘湯含藥血清組在H2O2處理前先用含藥血清預(yù)處理細(xì)胞；PI3K抑制劑組在H2O2處理前先用PI3K抑制劑LY294002預(yù)處理細(xì)胞；聯(lián)合用藥組則同時(shí)用含藥血清和LY294002預(yù)處理細(xì)胞。Theexperimentwasdividedintoanormalcontrolgroup,amodelgroup,aLingguiZhuganTangcontainingserumgroup,aPI3Kinhibitorgroup,andacombinationtherapygroup.Exceptforthenormalcontrolgroup,allothergroupsweretreatedwithH2O2attheoptimalmodelingconcentration.BeforeH2O2treatment,thecellsintheLingguiZhuganTangcontainingserumgroupwerepretreatedwiththecontainingserum;ThePI3KinhibitorgrouppretreatedcellswiththePI3KinhibitorLY294002beforeH2O2treatment;ThecombinationtherapygroupsimultaneouslypretreatedcellswithdrugcontainingserumandLY294按照CCK-8試劑盒說明書操作，用酶標(biāo)儀測(cè)定各孔在450nm處的吸光度值，計(jì)算細(xì)胞活力。AccordingtotheinstructionsoftheCCK-8reagentkit,measuretheabsorbancevaluesofeachwellat450nmusinganenzyme-linkedimmunosorbentassay(ELISA)readerandcalculatecellviability.6WesternBlot檢測(cè)PI3K/Akt信號(hào)通路相關(guān)蛋白表達(dá)WesternBlotdetectionofPI3K/Aktsignalingpathwayrelatedproteinexpression收集各組細(xì)胞，提取總蛋白，經(jīng)SDS電泳、轉(zhuǎn)膜、封閉后，分別用兔抗Akt、兔抗p-Akt抗體進(jìn)行孵育，再用HRP標(biāo)記的山羊抗兔IgG進(jìn)行二抗孵育，最后進(jìn)行化學(xué)發(fā)光顯色，用凝膠成像系統(tǒng)拍照并分析蛋白表達(dá)水平。Collectcellsofeachgroup,extracttotalprotein,andincubatewithrabbitantiAktandrabbitantip-AktantibodiesrespectivelyafterSDSelectrophoresis,membranetransferandsealing,thenincubatewithgoatantirabbitIgGlabeledwithHRPforsecondaryantibody,andfinallyconductchemiluminescencecoloration,takephotoswithgelimagingsystemandanalyzetheproteinexpressionlevel.所有數(shù)據(jù)均以均數(shù)±標(biāo)準(zhǔn)差（x±s）表示，采用SPSS軟件進(jìn)行統(tǒng)計(jì)分析。多組間比較采用單因素方差分析（One-wayANOVA），以P<05為差異有統(tǒng)計(jì)學(xué)意義。Alldataareexpressedasmean±standarddeviation(x±s)andanalyzedusingSPSSsoftware.Multiplegroupcomparisonswereconductedusingone-wayANOVA,withP<05indicatingstatisticallysignificantdifferences.三、結(jié)果Result為了探究苓桂術(shù)甘湯含藥血清對(duì)H2O2誘導(dǎo)的H9c2細(xì)胞損傷的保護(hù)作用，我們首先對(duì)細(xì)胞進(jìn)行了不同濃度的H2O2處理，并觀察了細(xì)胞的存活率。結(jié)果顯示，隨著H2O2濃度的增加，H9c2細(xì)胞的存活率逐漸降低。然而，在加入苓桂術(shù)甘湯含藥血清后，細(xì)胞的存活率得到了顯著提高，顯示出該藥物對(duì)H9c2細(xì)胞的保護(hù)作用。InordertoinvestigatetheprotectiveeffectofLingguiZhuganTangcontainingserumonH2O2induceddamagetoH9c2cells,wefirsttreatedthecellswithdifferentconcentrationsofH2O2andobservedtheirsurvivalrate.TheresultsshowedthatastheconcentrationofH2O2increased,thesurvivalrateofH9c2cellsgraduallydecreased.However,afteraddingtheserumcontainingLingguiZhuganTang,thesurvivalrateofcellswassignificantlyimproved,demonstratingtheprotectiveeffectofthedrugonH9c2cells.PI3K/Akt信號(hào)通路在苓桂術(shù)甘湯含藥血清保護(hù)H9c2細(xì)胞中的作用TheroleofPI3K/AktsignalingpathwayintheprotectiveeffectofLingguiZhuganTangcontainingserumonH9c2cells為了進(jìn)一步探討PI3K/Akt信號(hào)通路在苓桂術(shù)甘湯含藥血清保護(hù)H9c2細(xì)胞中的作用，我們檢測(cè)了處理后的細(xì)胞中PI3K/Akt信號(hào)通路的活性。結(jié)果顯示，在加入苓桂術(shù)甘湯含藥血清后，PI3K/Akt信號(hào)通路的活性得到了顯著增強(qiáng)。同時(shí)，我們還發(fā)現(xiàn)，當(dāng)使用PI3K/Akt信號(hào)通路的抑制劑時(shí)，苓桂術(shù)甘湯含藥血清對(duì)H9c2細(xì)胞的保護(hù)作用被明顯削弱。這些結(jié)果表明，PI3K/Akt信號(hào)通路在苓桂術(shù)甘湯含藥血清保護(hù)H9c2細(xì)胞中發(fā)揮了重要作用。InordertofurtherexploretheroleofthePI3K/AktsignalingpathwayintheprotectiveeffectofLingguiZhuganTangcontainingserumonH9c2cells,wedetectedtheactivityofthePI3K/Aktsignalingpathwayinthetreatedcells.TheresultsshowedthattheactivityofthePI3K/AktsignalingpathwaywassignificantlyenhancedaftertheadditionofLingguiZhuganTangcontainingserum.Meanwhile,wealsofoundthatwhenusinginhibitorsofthePI3K/Aktsignalingpathway,theprotectiveeffectofLingguiZhuganTangcontainingserumonH9c2cellswassignificantlyweakened.TheseresultsindicatethatthePI3K/AktsignalingpathwayplaysanimportantroleintheprotectionofH9c2cellsbyLingguiZhuganTangcontainingserum.活性氧（ROS）是細(xì)胞損傷的重要媒介，因此我們進(jìn)一步檢測(cè)了苓桂術(shù)甘湯含藥血清對(duì)H9c2細(xì)胞內(nèi)ROS水平的影響。結(jié)果顯示，在H2O2誘導(dǎo)下，H9c2細(xì)胞內(nèi)的ROS水平顯著升高。然而，在加入苓桂術(shù)甘湯含藥血清后，細(xì)胞內(nèi)的ROS水平得到了明顯降低。這表明苓桂術(shù)甘湯含藥血清可以通過降低細(xì)胞內(nèi)ROS水平來減輕H2O2對(duì)H9c2細(xì)胞的損傷。Reactiveoxygenspecies(ROS)areimportantmediatorsofcelldamage,sowefurtherinvestigatedtheeffectofLingguiZhuganTangcontainingserumonintracellularROSlevelsinH9c2cells.TheresultsshowedthatunderH2O2induction,thelevelofROSinH9c2cellssignificantlyincreased.However,afteraddingtheserumcontainingLingguiZhuganTang,theintracellularROSlevelwassignificantlyreduced.ThisindicatesthattheserumcontainingLingguiZhuganTangcanalleviatethedamageofH2O2toH9c2cellsbyreducingintracellularROSlevels.我們的研究結(jié)果表明苓桂術(shù)甘湯含藥血清可以通過激活PI3K/Akt信號(hào)通路并降低細(xì)胞內(nèi)ROS水平來保護(hù)H2O2誘導(dǎo)的H9c2細(xì)胞損傷。這為苓桂術(shù)甘湯在心血管疾病治療中的應(yīng)用提供了理論依據(jù)。OurresearchresultsindicatethatLingguiZhuganTangcontainingserumcanprotectH9c2cellsfromH2O2induceddamagebyactivatingthePI3K/AktsignalingpathwayandreducingintracellularROSlevels.ThisprovidesatheoreticalbasisfortheapplicationofLingguiZhuganTanginthetreatmentofcardiovasculardiseases.四、討論Discussion本研究主要探討了苓桂術(shù)甘湯含藥血清對(duì)H2O2誘導(dǎo)的H9c2細(xì)胞損傷的保護(hù)作用及其機(jī)制。通過實(shí)驗(yàn)結(jié)果，我們證實(shí)了苓桂術(shù)甘湯含藥血清可以顯著減輕H2O2對(duì)H9c2細(xì)胞的損傷，并揭示這一保護(hù)作用與PI3K/Akt信號(hào)通路的激活有關(guān)。ThisstudymainlyexplorestheprotectiveeffectandmechanismofLingguiZhuganTangcontainingserumonH2O2induceddamagetoH9c2cells.Throughexperimentalresults,wehaveconfirmedthattheserumcontainingLingguiZhuganTangcansignificantlyreducethedamageofH2O2toH9c2cells,andrevealthatthisprotectiveeffectisrelatedtotheactivationofthePI3K/Aktsignalingpathway.H2O2作為一種常見的氧化應(yīng)激誘導(dǎo)劑，被廣泛用于模擬細(xì)胞內(nèi)的氧化應(yīng)激環(huán)境。在本研究中，H2O2成功誘導(dǎo)了H9c2細(xì)胞的損傷，表現(xiàn)為細(xì)胞存活率的下降和細(xì)胞凋亡的增加。這與之前的報(bào)道一致，進(jìn)一步驗(yàn)證了我們的實(shí)驗(yàn)?zāi)Ｐ偷挠行?。H2O2,asacommonoxidativestressinducer,iswidelyusedtosimulatetheoxidativestressenvironmentwithincells.Inthisstudy,H2O2successfullyinduceddamagetoH9c2cells,characterizedbyadecreaseincellsurvivalrateandanincreaseincellapoptosis.Thisisconsistentwithpreviousreportsandfurthervalidatestheeffectivenessofourexperimentalmodel.苓桂術(shù)甘湯作為一種傳統(tǒng)中藥方劑，具有多種藥理作用，包括抗氧化、抗炎、抗凋亡等。本研究發(fā)現(xiàn)，苓桂術(shù)甘湯含藥血清可以顯著減輕H2O2誘導(dǎo)的H9c2細(xì)胞損傷，提高細(xì)胞存活率，并抑制細(xì)胞凋亡。這一結(jié)果與苓桂術(shù)甘湯的藥理作用相符，進(jìn)一步證實(shí)了其在細(xì)胞保護(hù)方面的有效性。LingguiZhuganTang,asatraditionalChinesemedicineformula,hasvariouspharmacologicaleffects,includingantioxidant,anti-inflammatory,andantiapoptoticeffects.ThisstudyfoundthatLingguiZhuganTangcontainingserumcansignificantlyalleviateH2O2induceddamagetoH9c2cells,improvecellsurvivalrate,andinhibitcellapoptosis.ThisresultisconsistentwiththepharmacologicaleffectofLingguiZhuganTang,furtherconfirmingitseffectivenessincellprotection.更重要的是，本研究還發(fā)現(xiàn)，苓桂術(shù)甘湯含藥血清的保護(hù)作用與PI3K/Akt信號(hào)通路的激活有關(guān)。PI3K/Akt信號(hào)通路在細(xì)胞生存、增殖和凋亡等方面發(fā)揮著重要作用。本研究發(fā)現(xiàn)，苓桂術(shù)甘湯含藥血清可以激活PI3K/Akt信號(hào)通路，進(jìn)而抑制細(xì)胞凋亡，保護(hù)H9c2細(xì)胞免受H2O2的損傷。這一發(fā)現(xiàn)為我們深入理解苓桂術(shù)甘湯的藥理作用提供了新的視角，也為進(jìn)一步開發(fā)其臨床應(yīng)用提供了新的思路。Moreimportantly,thisstudyalsofoundthattheprotectiveeffectofLingguiZhuganTangcontainingserumisrelatedtotheactivationofthePI3K/Aktsignalingpathway.ThePI3K/Aktsignalingpathwayplaysanimportantroleincellsurvival,proliferation,andapoptosis.ThisstudyfoundthattheserumcontainingLingguiZhuganTangcanactivatethePI3K/Aktsignalingpathway,therebyinhibitingcellapoptosisandprotectingH9c2cellsfromH2O2damage.ThisdiscoveryprovidesanewperspectiveforustodeeplyunderstandthepharmacologicaleffectsofLingguiZhuganTang,andalsoprovidesnewideasforfurtherdevelopingitsclinicalapplications.然而，本研究仍存在一定的局限性。我們僅從細(xì)胞層面探討了苓桂術(shù)甘湯含藥血清的保護(hù)作用及機(jī)制，未能在動(dòng)物或人體水平進(jìn)行驗(yàn)證。未來研究可以考慮構(gòu)建動(dòng)物模型或進(jìn)行臨床試驗(yàn)，以進(jìn)一步驗(yàn)證本研究的結(jié)論。本研究?jī)H關(guān)注了PI3K/Akt信號(hào)通路，未對(duì)其他可能涉及的信號(hào)通路進(jìn)行探討。未來研究可以進(jìn)一步拓展信號(hào)通路的研究范圍，以更全面地了解苓桂術(shù)甘湯的藥理作用機(jī)制。However,thisstudystillhascertainlimitations.WehaveonlyexploredtheprotectiveeffectandmechanismofLingguiZhuganTangcontainingserumatthecellularlevel,andhavenotbeenvalidatedattheanimalorhumanlevel.Futureresearchcanconsiderconstructinganimalmodelsorconductingclinicaltrialstofurthervalidatetheconclusionsofthisstudy.ThisstudyonlyfocusedonthePI3K/Aktsignalingpathwayanddidnotexploreotherpossiblesignalingpathwaysinvolved.FutureresearchcanfurtherexpandthescopeofsignalpathwayresearchtocomprehensivelyunderstandthepharmacologicalmechanismofLingguiZhuganTang.本研究證實(shí)了苓桂術(shù)甘湯含藥血清對(duì)H2O2誘導(dǎo)的H9c2細(xì)胞損傷的保護(hù)作用，并揭示了其與PI3K/Akt信號(hào)通路的關(guān)系。這些發(fā)現(xiàn)為苓桂術(shù)甘湯的藥理作用研究提供了新的依據(jù)，也為進(jìn)一步開發(fā)其臨床應(yīng)用提供了新的思路。然而，仍需進(jìn)行更深入的研究以驗(yàn)證本研究的結(jié)論并拓展其應(yīng)用范圍。ThisstudyconfirmstheprotectiveeffectofLingguiZhuganTangcontainingserumonH2O2induceddamagetoH9c2cellsandrevealsitsrelationshipwiththePI3K/Aktsignalingpathway.ThesefindingsprovidenewevidenceforthepharmacologicalstudyofLingguiZhuganTangandnewideasforfurtherdevelopingitsclinicalapplication.However,furtherresearchisstillneededtovalidatetheconclusionsofthisstudyandexpanditsapplicationscope.五、結(jié)論Conclusion本研究通過深入探究苓桂術(shù)甘湯含藥血清對(duì)H2O2誘導(dǎo)的H9c2細(xì)胞損傷的保護(hù)作用及其機(jī)制，發(fā)現(xiàn)苓桂術(shù)甘湯含藥血清能夠有效減輕H2O2引起的H9c2細(xì)胞損傷，并通過PI3K/Akt信號(hào)通路發(fā)揮其保護(hù)作用。ThisstudyinvestigatedindepththeprotectiveeffectandmechanismofLingguiZhuganTangcontainingserumonH2O2inducedH9c2celldamage.ItwasfoundthatLingguiZhuganTangcontainingserumcaneffectivelyalleviateH2O2inducedH9c2celldamageandexertitsprotectiveeffectthroughthePI3K/Aktsignalingpathway.實(shí)驗(yàn)結(jié)果顯示，在H2O2誘導(dǎo)的氧化應(yīng)激條件下，H9c2細(xì)胞活性明顯降低，細(xì)胞凋亡率顯著上升。然而，在加入苓桂術(shù)甘湯含藥血清后，細(xì)胞活性得到顯著提升，細(xì)胞凋亡率也明顯降低。這表明苓桂術(shù)甘湯含藥血清對(duì)H9c2細(xì)胞具有顯著的保護(hù)作用。TheexperimentalresultsshowedthatunderH2O2inducedoxidativestressconditions,theactivityofH9c2cellssignificantlydecreasedandtheapoptosisratesignificantlyincreased.However,afteraddingtheserumcontainingLingguiZhuganTang,thecellactivitywassignificantlyimprovedandtheapoptosisratewasalsosignificantlyreduced.ThisindicatesthattheserumcontainingLingguiZhuganTanghasasignificantprotectiveeffectonH9c2cells.進(jìn)一步的研究發(fā)現(xiàn)，苓桂術(shù)甘湯含藥血清能夠激活PI3K/Akt信號(hào)通路，提高Akt的磷酸化水平。這一發(fā)現(xiàn)揭示了苓桂術(shù)甘湯含藥血清保護(hù)H9c2細(xì)胞損傷的可能機(jī)制，即通過激活PI3K/Akt信號(hào)通路，抑制細(xì)胞凋亡，促進(jìn)細(xì)胞存活。FurtherresearchhasfoundthattheserumcontainingLingguiZhuganTangcanactivatethePI3K/AktsignalingpathwayandincreasethephosphorylationlevelofAkt.ThisdiscoveryrevealsapossiblemechanismbywhichtheserumcontainingLingguiZhuganTangprotectsH9c2cellsfromdamage,byactivatingthePI3K/Aktsignalingpathway,inhibitingcellapoptosis,andpromotingcellsurvival.本研究為苓桂術(shù)甘湯含藥血清在心血管疾病治療中的應(yīng)用提供了理論基礎(chǔ)。然而，仍需進(jìn)一步的研究來深入探討苓桂術(shù)甘湯含藥血清對(duì)PI3K/Akt信號(hào)通路的調(diào)控機(jī)制，以及其在心血管疾病治療中的具體療效和安全性。這些研究將有助于推動(dòng)苓桂術(shù)甘湯含藥血清在臨床實(shí)踐中的應(yīng)用，為心血管疾病的治療提供新的思路和方法。ThisstudyprovidesatheoreticalbasisfortheapplicationofLingguiZhuganTangcontainingseruminthetreatmentofcardiovasculardiseases.However,furtherresearchisneededtoexploretheregulatorymechanismofLingguiZhuganTangcontainingserumonthePI3K/Aktsignalingpathway,aswellasitsspecificefficacyandsafetyinthetreatmentofcardiovasculardiseases.ThesestudieswillhelppromotetheapplicationofLingguiZhuganTangcontainingseruminclinicalpractice,providingnewideasandmethodsforthetreatmentofcardiovasculardiseases.七、致謝Thanks我要向我的導(dǎo)師致以最深的敬意和感謝。在整個(gè)研究過程中，導(dǎo)師給予了我無私的指導(dǎo)與幫助，從選題、實(shí)驗(yàn)設(shè)計(jì)到論文撰寫，每一個(gè)環(huán)節(jié)都傾注了導(dǎo)師的心血和智慧。導(dǎo)師嚴(yán)謹(jǐn)?shù)目蒲袘B(tài)度、深厚的學(xué)術(shù)造詣以及高尚的師德，讓我受益匪淺，成為我人生道路上的重要財(cái)富。Iwouldliketoexpressmydeepestrespectandgratitudetomymentor.Throughouttheentireresearchprocess,mysupervisorprovidedmewithselflessguidanceandassistance,pouringmymentor'shardworkandwisdomintoeverystepfromtopicselection,experimentaldesigntop