富路韓穎

哈爾濱醫科大學附屬第一醫院心內科雄激素對心衰雄鼠心臟交感神經重構的影響

TheFirstClinicalCollegeofHarbinMedicalUniversityCardiacContractility交感神經系統在心力衰竭中的作用PeripheralPerfusionSympatheticActivationApoptosis、Necrosis、Fibrosis、Hypertrophy

?-Adrenergicdesensitization

SympatheticRemodelingCardiacRemodelingViciousCycle

TheFirstClinicalCollegeofHarbinMedicalUniversity

研究背景Androgenbeneficial?deleterious?女性CHF發病率死亡率男性Estrogenicprotection

TheFirstClinicalCollegeofHarbinMedicalUniversity研究背景DeficienciesinDHEAS,TT,eFTandIGF-1wereseenacrossallagecategoriesinmenwithCHFDHEAS,TTandeFTwereinverselyrelatedtoNYHAclassirrespectiveofcauseMenwithCHFandnormallevelofallanabolichormoneshadthebest3-yearsurvivalrate(83%)comparedwiththosewithdeficenciesin1(74%),2(55%),orall3(27%)anabolicendocrineaxes

TheFirstClinicalCollegeofHarbinMedicalUniversity

實驗目的

觀察內源性雄激素缺乏及生理劑量睪酮補充治療對慢性心力衰竭雄鼠心臟交感神經重構的作用實驗方法Wistar雄鼠60只（200±20g）

去勢/假去勢術后1w給予異丙腎340mg/kg,共2次正常組ControlS-Cas+PlaCas+PlaCas+TesCas+Tes+Flu術后予花生油2g/kg,1次/2d,共8周術后予花生油2mg/kg,1次/2d,共8周術后予睪酮2mg/kg,1次/2d,共8周術后予睪酮2mg/kg,1次/2d,氟他胺30mg/kg，1次/2d,共8周血流動力學心肌NE血漿NE免疫組化THNGFWesternTHNGF是NE合成的限速酶，其陽性表達可代表交感神經在心臟中的分布。是神經營養因子家族的代表，對交感神經的生長、分化、存活、突觸功能調控都有重要作用實驗結果

TheFirstClinicalCollegeofHarbinMedicalUniversity血流動力學結果組別例數HR,beats/min

LVSDP,mmHg

LVEDP,mmHg

LeftventriculardP/dt,mmHg/s

Control

10367±29

109.6±8.8

5.3±2.6

4587.6±566.8

S-cas+Pla

9414±25*87.5±9.3

*18.9±2.5

*△3352.0±639.9

*Cas+Pla

8425±22*

82.4±9.1*

23.7±2.7

*3064.3±594.7*

Cas+Tes

10408±21

92.3±9.6

*16.8±3.1

*△3467.7±435.0

*Cas+Tes+Flu

9404±26

88.7±10.1

*17.5±2.9

*△3212.0±618.6

*與Control組比較，*P<0.05；與Cas+HF+P組比較，△

P<0.05。

表1各組大鼠血流動力學指標比較

TheFirstClinicalCollegeofHarbinMedicalUniversity血漿及心肌NE*△**△*△△*△*△*△*

△與正常組比較，*P<0.05與Cas+HF+P組比較，△P<0.05

TheFirstClinicalCollegeofHarbinMedicalUniversity

心肌TH免疫組化染色

正常組S-cas+Pla組Cas+Pla組Cas+TesCas+Tes+Flu組

TheFirstClinicalCollegeofHarbinMedicalUniversity心肌TH免疫組化染色*△*△*△*△與正常組比較，*P<0.05與Cas+HF+P組比較，△P<0.05每組大鼠觀察兩張切片，每張切片計數5個（×200）陽性率評定標準為結合表達面積和染色強度綜合判定。

TheFirstClinicalCollegeofHarbinMedicalUniversity

心肌NGF免疫組化染色

正常組S-cas+Pla組Cas+Pla組Cas+Tes組Cas+Tes+Flu組

TheFirstClinicalCollegeofHarbinMedicalUniversity心肌NGF免疫組化染色與正常組比較，*P<0.05與Cas+HF+P組比較，△P<0.05每組大鼠觀察兩張切片，每張切片計數5個（×200）陽性率評定標準為結合表達面積和染色強度綜合判定。

**△*△*△△

TheFirstClinicalCollegeofHarbinMedicalUniversity心肌THWesternBlot

與正常組比較，*P<0.05；與Cas+HF+P組比較，△P<0.05TH60kDaGAPDH36kDaControlS-cas+Pla

Cas+Pla

Cas+Tes

Cas+Tes+Flu*△*△*△*△

TheFirstClinicalCollegeofHarbinMedicalUniversity心肌NGFWesternBlot

與正常組比較，*P<0.05；與Cas+HF+P組比較，△P<0.05NGF13kDaGAPDH36kDaControlS-cas+Pla

Cas+Pla

Cas+Tes

Cas+Tes+Flu△*△*△*△*討論

TheFirstClinicalCollegeofHarbinMedicalUniversity心衰時心肌壞死、纖維瘢痕形成→心臟交感神經軸索受其影響而出現功能障礙心衰時血漿NE作用于心肌細胞→心肌中NGF的含量降低→交感神經分布減少TH、NGF↓，心臟交感神經分布密度↓交感神經失支配動物心衰交感神經重構血漿NE

水平↑

，心肌NE含量↓心肌NE耗竭釋放NE↑再攝取NE

↓

TH↓，NE合成↓心衰交感神經過度激活心肌NE耗竭觸發反應遲鈍

TheFirstClinicalCollegeofHarbinMedicalUniversity本研究顯示心肌NE含量TH、NGF陽性神經密度心肌TH、NGF蛋白表達

去勢生理睪酮替代心臟交感神經重構

TheFirstClinicalCollegeofHarbinMedicalUniversityAndrogensignaltransductionTTargetcellNongenomiceffectsT5α-reductasearomataseDHTTE2ARERα/βGenomiceffectsCellfunctionGrowthDifferentiationProliferationapoptosis本實驗中應用AR阻斷劑未能阻斷睪酮的神經保護作用

TheFirstClinicalCollegeofHarbinMedicalUniversity結論

在慢性心力衰竭雄性大鼠，內源性睪酮水平下降加重了心功能不全，并加重心臟交感神經重構；

生理劑量睪酮替代治療可以改善大鼠的心功能和心臟交感神經重構。

這為睪酮補充治療男性慢性心衰的臨床可行性提供了理論依據。12

TheFirstClinicalCollegeofHarbinMedicalUniversityThankYou!LuFu,YingHan

CardiovascularDepartment,FirstAffiliatedHospital,HarbinMedicalUniversity,China

Theeffectsoftestosteroneuponcardiacsympatheticfunctioninratswithinducedheartfailure

TheFirstClinicalCollegeofHarbinMedicalUniversityCardiacContractilityThesympatheticsysteminCHFPeripheralPerfusionSympatheticActivationApoptosis、Necrosis、Fibrosis、Hypertrophy

?-Adrenergicdesensitization

SympatheticRemodelingCardiacRemodelingViciousCycle

TheFirstClinicalCollegeofHarbinMedicalUniversityBackgroundAndrogenbeneficial?deleterious?FemalesCHFmorbiditymortality

MalesEstrogenicprotection

TheFirstClinicalCollegeofHarbinMedicalUniversityBackgroundDeficienciesinDHEAS,TT,eFTandIGF-1wereseenacrossallagecategoriesinmenwithCHFDHEAS,TTandeFTwereinverselyrelatedtoNYHAclassirrespectiveofcauseMenwithCHFandnormallevelofallanabolichormoneshadthebest3-yearsurvivalrate(83%)comparedwiththosewithdeficenciesin1(74%),2(55%),orall3(27%)anabolicendocrineaxes

TheFirstClinicalCollegeofHarbinMedicalUniversityNeuroprotectiveeffectsoftestosterone

Thishormonehasbeenlinkedtoanincreaseinneuronalsomalsize,neuriticgrowth,plasticityandsynaptogenesisinbothmotoneuronsofthespinalnucleusofthebulbocavernosusandseveralpopulationsofpelvicautonomicneurons

Maleshavehighersympatheticcardiacautonomicactivitythanfemales.Post-infarctionarrhythmiasinducedbysympatheticactivationaremoresevereinmalethanfemalerats.

Testosteroneactivatescholineacetyltransferaseandtyrosinehydroxylase(TH)andinfluencesNEmetabolism,storage,andrelease

Testosteroneisinvolvedwithnervoussystemdevelopment.Someresearchhasshownthatandrogenhasaneuroprotectionagainsthypoxia-ischemiabrain

damage.

TheFirstClinicalCollegeofHarbinMedicalUniversity

Aim

Thecurrentstudywasdesignedtodeterminewhethercastrationwithorwithouttestosteronereplacementresultedinchangesincardiacsympatheticnerveactivityinratswithinducedheartfailure.

Methods60Wistarmalerats（200±20g）

1w

aftercastration/sham-castration,ratsreceivedasubcutaneousinjectionofisoproterenolat340mg/kg,atotaloftwoinjections

ControlS-Cas+PlaCas+PlaCas+TesCas+Tes+Flupeanutoil2g/kg,1/d,for8weekspeanutoil2g/kg,1/d,for8weekstestosterone

2mg/kg,1/d,for8weekstestosterone

2mg/kg,flutamide

25mg/kg,1/d,for8weeksMyocardialNEplasmaNEImmunohisto-chemistry

THNGFWesternBlotanalysis

THNGFrestinghemodyna-micsResults

TheFirstClinicalCollegeofHarbinMedicalUniversityResultsofrestinghemodynamics

*P<0.05vscontrolgroup；△

P<0.05

vsCas+Plagroup。

Table1resultsof

restinghemodynamics

groupnHR,beats/min

LVSDP,mmHg

LVEDP,mmHg

LeftventriculardP/dt,mmHg/s

Control

10367±29

109.6±8.8

5.3±2.6

4587.6±566.8

S-cas+Pla

9414±25*87.5±9.3

*18.9±2.5

*△3352.0±639.9

*Cas+Pla

8425±22*

82.4±9.1*

23.7±2.7

*3064.3±594.7*

Cas+Tes

10408±21

92.3±9.6

*16.8±3.1

*△3467.7±435.0

*Cas+Tes+Flu

9404±26

88.7±10.1

*17.5±2.9

*△3212.0±618.6

*

TheFirstClinicalCollegeofHarbinMedicalUniversitytheplasmaandcardiacNEcontents

*△**△*△△*△*△*△*

△*P<0.05vscontrolgroup△P<0.05vsCas+Plagroup

TheFirstClinicalCollegeofHarbinMedicalUniversity

ImmunohistochemicalstainingforTH

ControlgroupS-cas+PlagroupCas+PlagroupCas+TesgroupCas+Tes+Flugroup

TheFirstClinicalCollegeofHarbinMedicalUniversitytheTH-positivenervecelldensity

*△*△*△*△ThedensityofTH-labelednervefiberswasquantitativelyestimatedfrom10randomlyselectedfieldsatamagnificationof×200.

Thenervedensitywasmeasuredontracingsbycomputerizedplanimetry.

*P<0.05vscontrolgroup△P<0.05vsCas+Plagroup

TheFirstClinicalCollegeofHarbinMedicalUniversity

Immunohistochemicalstainingfor

NGFControlgroupS-cas+PlagroupCas+PlagroupCas+TesgroupCas+Tes+Flugroup

TheFirstClinicalCollegeofHarbinMedicalUniversitytheNGF-positivenervecelldensity

與正常組比較，*P<0.05與Cas+HF+P組比較，△P<0.05**△*△*△△ThedensityofTH-labelednervefiberswasquantitativelyestimatedfrom10randomlyselectedfieldsatamagnificationof×200.

Thenervedensitywasmeasuredontracingsbycomputerizedplanimetry.

TheFirstClinicalCollegeofHarbinMedicalUniversity

theexpressionofTHprotein

*P<0.05vscontrolgroup；△

P<0.05

vsCas+Plagroup。TH60kDaGAPDH36kDaControlS-cas+Pla

Cas+Pla

Cas+Tes

Cas+Tes+Flu*△*△*△*△

TheFirstClinicalCollegeofHarbinMedicalUniversitytheexpressionofNGFprotein

*P<0.05vscontrolgroup；△

P<0.05

vsCas+Plagroup。NGF13kDaGAPDH36kDaControlS-cas+Pla

Cas+Pla

Cas+Tes

Cas+Tes+Flu△*△*△*△*Discussion

TheFirstClinicalCollegeofHarbinMedicalUniversityMyocardialischaemiaandcelldeath→cardiacsympatheticnervefiberdysfunctionPlasmaNEaffectsmyocardium→NGF↓

→thedensityofcardiacsympatheticnerve↓

TH、NGF↓，thedensityofcardiacsympatheticnerves↓cardiacsympatheticremodeling

SympatheticRemodelinginCHFplasma

NE↑

，myocardialNE↓exhaustofmyocardialNENErelease↑NE

reuptake↓

TH↓NE

synthesis

↓exhaustofmyocardialNEover-activationofcardiacsympatheticnervoussystem

TheFirstClinicalCollegeofHarbinMedicalUniversityWhatthereasearchtellusmyocardialNEcontentTH/NGF

positivenervedensityTH/NGF

proteinexpressionCastrationTes