山莨菪碱对利血平大鼠胃粘膜损伤的影响精Word格式文档下载.docx

山莨菪碱对利血平大鼠胃粘膜损伤的影响精Word格式文档下载.docx

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一氧化氮

中国病理生理杂志000313

[摘要]目的：

研究山莨菪碱对利血平大鼠胃粘膜损伤的影响及其作用机制。

方法：

利用利血平大鼠胃粘膜损伤模型，观察腹注山莨菪碱对利血平大鼠胃粘膜损伤、胃酸分泌、胃粘液分泌、胃运动、胃粘膜血流量及胃粘膜一氧化氮含量和一氧化氮合成酶活性变化的影响。

结果：

山莨菪碱能抑制利血平大鼠胃粘膜损伤灶的形成；

山莨菪碱能抑制利血平大鼠胃酸分泌，但对胃液分泌的量无影响；

山莨菪碱能促进利血平大鼠胃粘液分泌和胃粘膜血流量，抑制胃的运动；

山莨菪碱能抑制利血平导致的大鼠胃粘膜一氧化氮含量的降低和一氧化氮合成酶活性的降低。

结论：

山莨菪碱抑制利血平大鼠胃粘膜损伤的形成与其抑制胃酸分泌、胃的运动，促进胃粘液分泌、增加胃粘膜血流量有关；

NO可能在介导山莨菪碱抑制利血平大鼠胃粘膜损伤形成中有重要作用。

[中分类号]R961R961[文献标识码]A

[文章编号]1000-4718（2000）03-0237-06

Effectofanisodamineonthereserpine-inducedgastricmucosallesioninrats

WANJun-li

（DepartmentofBiology,YantaiTeachersCollege,Yantai264025，China）

[Abstract]AIM:

Todeterminetheeffectsofanisodamine（Ani）administeredintraperitoneallyonthegastricmucosallesioninducedbyreserpine.METHODS:

Inreserpine-treatedrats,gastricmucosallesion,gastricacidsecretion,gastricbarriermucussecretion,gastriccontraction,gastricmucosalbloodflow（GMBF）,gastricmucosalnitricoxidesynthase（NOS）activityandnitricoxide（NO）contentwereexamined.RESULTS:

Aniindosesof1,5and10mg/kgsignificantlyinhibitedtheformationofgastriclesionsinducedbyreserpine,withthesuppressiverateof60.0%,66.7%and76.6%,respectively.Ani（10mg/kg）significantlyinhibitedthesecretionofgastricacid,buthadnoeffectonthevolumeofgastricjuice.Ani（10mg/kg）significantlypromptedthesecretionofgastricbarriermucus.OurfindingsalsoshowedthatAni（10mg/kg）significantlysuppressedthefrequencyandamplitudeofgastriccontraction.Ani（10mg/kg）significantlypromptedGMBF.Inreserpinetreatedrats,gastricmucosalNOSactivityandNOcontentweredecreasedandAni（10mg/kg）couldinhibitthedecreaseinNOSactivityandNOcontent.CONCLUSIONS:

TheprotectiveeffectofAnimayresultsinpartfrominhibitinggastricacidsecretion,promptinggastricbarriermucussecretion,suppressinggastriccontractionandimprovingGMBF.NOseemstoplayanimportantmediatorroleintheAniprotectivemechanisms.

[MeSH]Gastricmucosa;

Reserpine;

Nitricoxide

[CLCnumber]R961[Documentcode]A

INTRODUCTION

Anisodamine（Ani）isanalkaloidinitiallyisolatedfromAnisodustanguticusPaschinChina.Asananalogofatropin,Anihasnumerouspharmacologicaleffectssuchasantiacetylcholine,antishock,improvingmicro-circulation,inhibitingplateletaggregation,antiarrhythmiaandcalciumantagonisticaction.Ithasbeenextensivelyusedtotreatendotoxinshock,spasmofgastrointestinalandothervisceralsmoothmuscleetc.

Yongetal[1]reportedthatAniadministeredorallywasfoundtoantagonizethegastricmucosaldamageinducedbyindomethacin,restraint,pyloricligationorabsoluteethanolingestioninrats.OurpreviousstudydemonstratedthatintraperitonealAniprotectedtheratsfromcontractinggastriclesioninducedbyrestraintwater-immersion[2].TheeffectofAniongastriclesioninducedbyreserpine,aswellasitsunderlyingmechanism,however,remainstobestudied.

Recentstudieshaveshownthatreductioninendogenousnitricoxide（NO）seemstoberesponsibleforthegastricmucosalinjuryinducedbyethanol[3],ischemia-reperfusion[4]andcoldrestraintstress[5].Nevertheless,whetherNOreductionisimplicatedinthereserpine-inducedgastriclesionhasnotyetbeeninvestigated.

Therefore,theaimsofthisstudyaretoinvestigate:

（1）whetherornotAniadministeredintraperitoneallycanprotectagainstgastriclesioninducedbyreserpineinrats,

（2）ifAnidoesgiveprotection,bywhatmechanisms?

and（3）thepossibleimplicationofNOintheAniprotection.

MATERIALSANDMETHODS

1.AnimalpreparationSprague-Dawleyratsofeithersex（150～220g），keptinindividualcageswitharaisedmeshbottomeach,weredeprivedoffoodbutallowedfreeaccesstowaterfor24hpriortotheexperiments.

2.Experimentalprocedure

ExperimentI.ProductionofgastriclesionandmeasurementofgastriclesionindicesTheanimalsweredividedintofourgroups.GroupI（control）receivedsaline（5mL/kg）intraperitoneally.GroupⅡ，Ⅲ,andⅣreceivedAniindosesof1,5and10mg/kgintraperitoneally,respectively.Thirtyminuteslater,theratsweregivenreserpineinadoseof5mg/kgforeachanimalintraperitoneally.Therodentswerekilledbycervicaldislocation6hafterthereserpineinjection.Theirstomachswereimmediatelyremoved,inflatedbyinjecting10mLof10%formalinandimmersedin10%formalinfor10min.Thestomachswerethenopenedalongthegreatercurvature.Thesumofthelengthofeachlesionwasexpressedasgastriclesionindex（mm）,asdescribedbyGuthetal.

ExperimentⅡ.MeasurementofgastricacidsecretionGastricacidsecretionwasmeasuredinpylorus-ligatedrats.Theanimalsweredividedintothreegroups.GroupI:

salinegroup;

groupⅡ：

reserpinegroup;

groupⅢ：

Anigroup.Saline（5mL/kg）wasgivenintraperitoneallyingroupⅠandⅡ；

Ani（10mg/kg）wasgivenintraperitoneallyingroupⅢ，respectively.Thirtyminuteslater,thepyloricendsofthestomachsweretiedoffunderetheranesthesia.Immediatelythereafter,reserpine（5mg/kgforeachrat）wasadministeredintraperitoneallyingroupⅡandⅢ,andsalinewasgiveningroupI.Sixhoursafterpylorusligation,theanimalswerekilledbycervicaldislocation,thenthestomachswereremovedandthegastriccontentswereseparatelydrainedintograduatedcentrifugetubes.Aftercentrifugationat3000r/minfor10min,thesupernatantswerecarefullydecantedandassayedforthevolumeofthegastricjuiceanditsH+concentration.TheH+concentrationwasdeterminedbytitratingthegastricjuicetopH7.0with0.01mol/LNaOH.Volumesofgastricsecretion,totalacidoutputandtitratableaciditywereexpressedasmLofgastricjuice/100gbodyweight,μmolofH+/100gbodyweightandmmolofH+/L,respectively.

ExperimentⅢ.MeasurementofgastricbarriermucusGastricbarriermucuswasdeterminedaccordingtothemodifiedprocedureofBoltonetal.TheanimalsweredividedintothreegroupsandtreatedasinexperimentⅡ.Sixhoursafterreserpineinjection,theanimalswerekilledandthestomachsremoved.Thestomachswereopenedalongthelessercurvatureandfreemucuswasgentlyremovedfromthesurfaceofthegastricmucosa.Eachofthestomachswasthenplacedseparatelyin20mLAlcineBluesolution（20mgAlcianBluewasdissolvedin100mLMcIlvainebuffer,pH5.8）andincubatedfor2hat20℃,followedbycentrifugationat4000r/minfor10minandthesupernatantswereusedformeasuringabsorbanceat615nmonaspectrophotometer.Gastricbarriermucuswascalculatedusingthefollowingformula:

gastricbarriermucus（mg）=4-4×

（testabsorbance）/（standardabsorbance）

Thecontentofgastricbarriermucuswasexpressedasmg/perstomach.

ExperimentⅣ.MeasurementofgastriccontractionGastriccontractionwasrecordedaccordingtothemethoddescribedbyTakeuchietal.Aminiaturegastricballoon（5mmindiameter）fashionedfromcondomrubberwasconnectedtooneendofapolyethylenetubewithtwosideholesonthisend.Theairintheballoonwasdrivenoutbyfillingitwithwater.Thewaterwasallowedtoescapetozeropressure,atwhichtimetheflaccidballooncontainedapproximately0.3mLofwater.Underetheranesthesia,anincisionontheabdominalwalloftheratwasmadeandtheballoonwasinsertedthroughacauteryholeintothegreatercurvatureoftheforestomachabout5mmfromthelimitingridge.Theballoonwasthentiedinplacesothatitlayintheglandularpartofthestomach,carebeingtakennottodamagetheballoonwiththetube.Theballoonandtubesystemwasconnectedtoapressuretransducerwhichwasjoinedtoaphysiologicalrecorder.Thentheincisionontheabdominalwallwassuturedwithsilk.Ratsweredividedintotwogroups.GroupⅠ:

groupⅡ:

Ani（10mg/kg）group.SalineorAniwasgivenintraperitoneally,respectively.Thirtyminlater,reserpinewasgivenintraperitoneallyinallratsandgastriccontractionswererecordedforatotalof6h.Allwaveswithamplitudesgreater4cmH2Oandlastinglongerthen2swerenotedascontractions.Frequencyofcontractionandamplitudeofcontractionwereanalyzedina1-hrecordingblock.

ExperimentV.MeasurementofgastricmucosalbloodflowRatsweredividedandtreatedasinexperimentⅡ.Gastricmucosalbloodflow（GMBF）wasmeasuredusingneutralredclearancetechniqueaccordingtotheproceduremodifiedbyZhanetal.Briefly,animalswereanesthetizedwithurethanegivenintraperitoneally（1.25g/kg）.Saline（5mL/kg）wasgiveningroupIandⅡ，Ani（10mg/kg）wasgiveningroupⅢ.Thirtyminuteslater,thepyloricendsofstomachsweretiedoff.Afterabolusdoseof1mLneutralredwasgivenintravenously,constantplasmaneutralredlevelwasmaintainedbycontinousintravenousinfusionofneutralred（3mg*kg-1*h-1）atarateof4mL/h,andthereaftersalinewasadministeredingroupIandreserpine（5mg/kg）wasgivenintraperitoneallyingroupⅡandⅢimmediately,respectively.Sixhourslater,samplesofblood（1mL）andgastriccontentwerecollected.ThecontentofneutralredintheplasmaandgastricjuiceweremeasuredandGMBFwerecalculatedaspreviouslydescribedbyZhan.TheresultswereexpressedasmL/min.

ExperimentⅥ.MeasurementofgastricmucosalNOSactivityandNOcontentTheanimalsweredividedandtreatedasinexperimentⅡ.Sixhoursafterreserpineinjection,theanimalswerekilledbycervicaldislocationandthestomachswereremoved.Thestomachswereopenedalongthegreatercurvatureandwashedwithcoldsaline.Themucosallayerwasseparatedfromthemuscularlayerwithasharpblade,themucosalmaterialwasweighe