(2)高倍光学显微镜下观察HE切片,空白对照组、氯氮平2mg/kg组、氟伏沙明2mg/kg组、氯氮平2mg/kg+氟伏沙明2mg/kg组、氯氮平10mg/kg+氟伏沙明2mg/kg组小鼠的胰岛均位于胰腺外分泌部之中,其岛型丰满、轮廓清晰、胰岛细胞均匀分布,均未见淋巴细胞浸润;尽管氯氮平10mg/kg组小鼠胰岛位于胰腺外分泌部
之中,其岛型丰满、轮廓清晰、胰岛细胞均匀分布,未见淋巴细胞
浸润,但是该组有2只小鼠的胰岛周围有淋巴细胞浸润,但胰岛内无淋巴细胞浸润。
(3)空白对照组、氯氮平2mg/kg组、氯氮平10mg/kg组、氟伏
博士学位论文中文摘要
沙明2mg/kg组、氯氮平2mg/kg+氟伏沙明2mg/kg组、氯氮平10mg/kg+氟伏沙明2mg/kg组胰岛细胞分布均匀,p细胞胞浆呈棕黄色胰岛素阳性表达,未见p细胞凋亡。
(4)小鼠GLUT4mRNA相对表达量与小鼠空腹血糖、去甲氯氮平血药浓度的值呈负相关(PO.05,r=0.347"--'0.555)。
四结论
连续28d给雄性C57BL/6小鼠腹腔注射10mg/kg氯氮平,
可以降低小鼠体重及高密度脂蛋白水平(皿L.C),升高小鼠空腹
血糖、糖耐量及胰岛素水平。
研究发现:
单独给予2mg/kg氟伏沙明治疗没有上述作用,氟伏沙明(2mg/kg)联合10mg/kg氯氮平治疗可以改善10mg/kg氯氮平对血糖、血脂的不良影响。
此结果为临床上氯氮平和氟伏沙明联合使用提供理论依据。
氟伏沙明与氯氮平联合使用可以抑制氯氮平的代谢,使去甲氯氮平的血浓度明显降低。
研究发现:
小鼠的空腹血糖与去甲氯氮平的浓度没有显著性相关,但有随氯氮平血药浓度升高而升高的趋势。
这提示:
氟伏沙明可以降低小鼠血浆中去甲氯氮平的血浓度,这可能是其与氯氮平联合使用可以改善氯氮平对血糖的升高作用的重要机制之一。
此结果为临床上使用氯氮平患者出现血糖升高的个体差
博士学位论文中文摘要
异性提供理论依据。
2mg/kg氯氮平和10mg/kg氯氮平可以降低小鼠股四头中GLUT4mRNA的表达。
研究发现:
氟伏沙明(2mg/kg)与氯氮平联合使用可以改善氯氮平对GLUT4mRNA表达的抑制作用;在小鼠体内GLUT4mRNA相对表达量与空腹血糖、去甲氯氮平血药浓
度的值呈负相关。
氟伏沙明、不同剂量的氯氮平对胰腺13细胞的凋亡没有影响,说明氟伏沙明、氯氮平对胰腺13细胞没有明显的毒性作用。
上述结果部分阐明了氯氮平升高血糖的机制。
关键词:
氯氮平,氟伏沙明,血糖,葡萄糖转运蛋白4,细胞凋亡
博士学位论文英文摘要
EFFECTSOFATYPICALANTIPSYCHOTICSCLOZAPINEONTHEBLOODGLUCOSEINTHEMALEC57BL/6MICEANDITSⅣ匝CHANISMRESEARCH
ABSTRACT
oBJECTIVE
Tostudytheeffectsofcoadministrationofclozapineandfluvoxamineversusclozapinemonotherapyonthemetabolismandinvestigatethemechanismofclozapine’hyperglycemiainthemaleC57BL/6mice.
METHoDS
1.TheeffectsofcoadministrationofelozapineandfluvoxamineversusclozapinemonotherapyonthemetabolisminthemaleC57BL/6mice
120maleC57BL/6miceweredividedinto6groups,
(1)vehicle,
(2)
clozapine2mg/kg,(3)clozapine10mg/kg,(4)fluvoxamine2mg/kg,
(5)clozapine2mg/kg+fluvoxamine2mg/kg,(6)clozapine10mg/kg+fluvoxamine2mg/kg.Themiceweretreatedwithcorrespondingdrugintraperitoneally(IP)QDat8:
30AMfor28consecutivedays.Bodyweightwasrecordedondays0,7,14,21,and28.Toassessglucosetolerance,miceunderwentovemightfastingonday29,andtheirbloodglucoseresponsestoIPadministrationof25%glucose(2g/kg)weredetermined.Bloodglucoseconcentrationwereassessedinthewhole
bloodcollectedfromthetailveinbeforeglucoseadministration(ie,time
博士学位论文英文摘要
0,FBG)and30,60,and120minutesafterglucoseadministration.Themicewerefastedfor6hoursafterglucoseadministrationandwereobtained0.6mLbloodbyremovingthemice’eyeballafteranesthetizedusingsodiumpentobarbital130mg/kg.Thebloodsampleswereimmediatelyplacedinheparin-coatedtubesforplasmalipid(includingtriglyceride,totalcholesterol,highdensitylipoproteincholesterolandlowdensitylipoproteincholester01),insulindeterminationandplasmadrugconcentration.
2.Theeffectsofeoadministrationfluvoxamineontheplasma
clozapineanditsmetabolitesconcentration
Theplasmaclozapineanditsmainmetabolites(desmethylclozapine
andclozapineN—oxide)concentrationweredeterminedinthefollowingexperimentgroup:
(2)clozapine2mg/kg,(3)clozapine10mg/kg,(5)
clozapine2mg/kg+fluvoxamine5mg/kgand(6)clozapine10mg/kg+fluvoxamine2mg/kg.Thenumberofplasmadeterminedineverygroupwasten.Thecorrelationbetweentheplasmaclozpineanditsmetabolites
(desmethylclozapineandclozapineN-oxide)concentration,plasmalipid
andfastedbloodglucosewasanalyzed.
Ahighperformanceliquidchromatographyelectrospraymass
spectrometry(HPLC-MS/ESI)methodstosimultaneouslydetermineclozapineanditsmetabolitesdesmethyl—clozapineandclozapine-N-oxideinmiceplasma.TheextractedsamplewasperformedonliquidchromatographyusingaWatersXtera刑MSC18column(150mmx3.9
mln,5grn)withcolumntemperature40。
C.Mobilephaseconsistedof
acetonitrileandwater(36:
64,includinglOmmol/Lammoniumacetateand0.01%formicacid,pH=5.96)withaflowrate1.0mL/min.Thecompoundswereionizedintheelectrosprayionization(ESI)ionsource,
.TX.
fragmentedbyin—sourcecollisionsandtheionizedmolecularandfragmentionsdetectedintheselectedionrecording(SIR)mode.ThesampleswereevaporatedbyN2anddissolvedmobilephaseandsamplesizewere209L.
3.ThemechanismofclozapineOHthebloodglucoseinthemaleC57BL/6mice
MaleC57BL/6miceweresacrificedbyceruicaluertebraluxationafterobtainingplasmabymovingmice’eyeball.ThenpancreaticsalivaryglandandmusculusquadricepsfexorisofC57BL/6
miceweretakenout.Pancreaticglandwereplacedinpercent10neutral
formaldehyde,fixed,imbeddedbyparaffinandcutparaffinsection(twoportion,thickness4um).MusculusquadricepsfexorisofC57BL/6miceweredeepfreezedbyplacingliquidnitrogenandtransferredinto一70。
C
refrigeratoruntildetermination.
(1)GLUT4mRNAexpressioninthemicemusculusquadricepsfexorisweredeterminedbyreversetranscriptionpclymerase(RT—PCR).
(2)Aparaffinsectionofmicepancreaticglandwereroutinely
deparaffinaged,stainedbyhematine—cosine(HE),andobservedwhether
theyhavetheinsulitis.
(3)Aparaffinsectionofmicepancreaticglandwereroutinelydeparaffinaged.Apoptosispcellweremarkedbyinsitunickend—labeling(TUNEL),pcellwerelabeledbystreptavidin—biotin-peroxidasecomplex(SABC)andthecountofpcellapoptosiswerecounted.
(4)ThecorrelationwereanalysedbetweentheexpressionofGLUTmRNA,thecountofpancreas13cellapoptosis,thedegreeofpancreatic
.X.
博士学位论文英文摘要
islandlymphocyteinfiltrateandfastedbloodglucose,plasmalipid,
plasmaclozapineanditsmetabolitesconcentration.RESULTS
1.ResultsofeffectsofcoadministrationofclozapineandfluvoxamineversusclozapinemonotherapyonthemetabolisminthemaleC57BL/6mice
Therewerenosiginificanceinbodyweightandfastedbloodglucoseamongtrialgroupsbeforeexperiment(尸>O.05).Thebodyweightsignificantlydecreasedinthe10mg/kgclozapine—treatedgroupsatdays14,21,and28(all,Pgroup.Inthe(2mg/kgclozapine-I-10mg&gfluvoxamine)-treatedgroup,bodyweightsignificantlyincreasedcomparedwiththevehicle-treatedgroup(尸O.05).Intheclozapine10mg/kggroup,bloodglucoseconcentrationsignificantlyincreased0,30,60,and120minutesafterglucoseadministration(all,P<0.05).Theclozapine10mg/kggroupalsohadsignificantincreaseinplasm