Cov teebmeem ntawm Cistanche Deserticola Extract Ntawm Noov erectile teb hauv Castrated nas

Abstract:Cistanche deserticola(CD) tau raug suav hais tias yog tus neeg sawv cev tonic ntawm kev ua me nyuam rau ntau txhiab xyoo. Cov teebmeem ntawm CD extract raunoov erectilecov lus teb tau raug tshawb xyuas hauv txoj kev tshawb fawb tam sim no. Tom qab kev phais castration, nas tau kho intragastrically nrog CD extract ({0}}.45, 0.90 thiab 1.8 g / kg) txhua hnub rau plaub lub lis piam.Noov erectileCov lus teb tau ntsuas thiab cov tshuaj hormones hauv cov ntshav tau kuaj thaum kawg ntawm qhov kev sim. Nws tau raug soj ntsuam tias coverectilelatencyua ntev thiab coverectile ntevluv luv nyob rau hauv castrated nas piv rau sham-operatedcontrols. Txawm li cas los xij, CD extract shortened luberectile latencythiab ncua lub sijhawm erectile kom txo qis qhov tsis zoo ntawm castration. Ntawm qhov ntau npaum li cas ntawm 0.9g / kg, CD extract tau tswj cov ntshav luteinizing hormone concentration mus rau theem ib txwm nyob hauv cov nas tsuag. Cov kev tshawb pom no qhia tau hais tias CD pab txhawb qhov noov erectile teb thiab hloov kho cov qib tshuaj hormones rau qee yam.

Ntsiab lus:Cistanche deserticola; erectile teb; castration; tshuaj hormone

Yog xav paub ntxiv txog cistanche txhim kho kev sib deev muaj nuj nqi

Taw qhia

CistancheHoffmg. Et Link, ib tug ntawm cov genera ntawm Orobanchaceae tsev neeg, feem ntau yog faib nyob rau hauv lub aridlands thiab suab puam nyob rau hauv sab qaum teb hemisphere, xws li Inner Mongolia,Xinjiang, Ningxia autonomous regions ntawm Tuam Tshoj, thiab Iran, Is Nrias teb, Mongolia, thiab lwm yam Cistanchespecies muaj nyob rau hauv perennial parasite tshuaj ntsuab, uas feem ntau xa mus rau cov hauv paus hniav ntawm xuab zeb-kho nroj tsuag xws li Haloxylox ammodendron, H. persicum, Kalidium foliatumand Tamarix nroj tsuag, thiab lwm yam. thiab Tu 2009).Herba Cistanche, lub qia ntawm Cistanche hom, tau raug suav hais tias yog ib tug superior tonic thiab tau txais lub meej mom ntawm "Ginseng ntawm cov suab puam". Ntawm Cistanche hom, Cistanche deserticola YC MA tau qhia tias yog cov khoom siv tseem ceeb ntawm Cistanche (Stefanova li al.2011). Txawm li cas los xij, vim muaj kev sau qoob loo ntau dhau xyoo tas los no, cov peev txheej ntawm cov tsiaj qus C. deserticola (CD) tsis txaus uas tau sau los ua ib qho ntawm Class IIplants xav tau kev tiv thaiv hauv Suav teb, yog li lwm hom tsiaj ntawm no genus, xws li C. salsa. thiab C. sinensis, kuj yog siv los hloov hauv qee qhov chaw. Los ntawm thawj zaug sau tseg hauv ShenNong's Suav Cov Khoom Siv Medica (ib qho khoom siv tshuaj qub txeeg qub teg), CD tau xav tias ua kom lub raum ua kom lub raum thiab txhawb lub "Yang" hauv kev kho mob raws li cov tshuaj suav tshuaj suav tshuaj, uas tau liam tias ua tau zoo rau kev yug me nyuam, kev loj hlob thiab fertility muaj nuj nqi. Lub caij no, CD kuj tau muab cov nqaij nyug, qos yaj ywm los yog nplej los tsim cov khoom noj tonic los kho kev puas tsuaj rau ntau pua xyoo, qhia tias nws muaj kev nyab xeeb rau kev noj qhov ncauj (Xiong li al.2013). Txawm li cas los xij, cov kev siv tsis zoo no feem ntau muaj nyob hauv Suav incunabula, nrog cov ntaub ntawv tshawb fawb tsis tshua muaj kev sib tham txog cov tswv yim tshuaj niaj hnub no.Erectile dysfunction (ED) yog ib qho mob tshwm sim los ntawm qhov tsis txaus erection ntawm noov lossis erection uas tsis kav ntev txaus rau cov txiv neej laus (Cohan 200Koren). ). Nyob rau hauv teb rau niaj hnub, sophisticated, tshuaj neeg sawv cev, muaj ib tug tshiab txaus siab mus tshawb fawb tshuaj zoo thiab kev nyab xeeb los ntawm cov nroj tsuag rau kev kho mob ntawm ED (Adimoelja 2000). Ib txoj kev tshawb fawb yav dhau los (Liuet al. 2009) tau qhia tias CD electuary accelerate qhov noov erection hauv cov nas ib txwm muaj, txawm li cas los xij, tsis muaj cov ntaub ntawv cuam tshuam txog kev cuam tshuam ntawm CD ntawm erectile kawg.Hauv tsiaj, phais castration nrog ob sab orchiectomyhas tau suav tias yog tus qauv ntawm castration. forinvestigating ED (Tombal 2005). Yog li, qhov kev tshawb fawb tam sim no tau ua los ntsuas qhov muaj peev xwm cuam tshuam ntawm CD tiv thaiv ED los ntawm kev tshawb xyuas qhov erectile latency thiab sijhawm ntev hauv cov nas tsuag.

Cov khoom siv thiab cov txheej txheem

 Chemicals and materials Echinacoside (purity >98%) and acteoside (purity >98 feem pua) tau yuav los ntawm National Institutes for Food and Drug Control (Beijing, China). HPLC qib methanol yog yuav los ntawm Sigma-Aldrich (St. Louis, MO, USA). Cov dej huv huv tau tsim los siv MilliporeMilli-Q Integral 3 System thiab Q-POD Milli-Q System (Bedford, MA, USA). Cov stems ntawm C. deserticola tau yuav los ntawm Shenzhen GURU Biology Co., Ltd (GuangDong, Suav teb) thiab hauv av rau hauv cov hmoov zoo nrog apulverizer dhau los ntawm 60mesh sieve.HPLC tsom xam thiab cov khoom siv cog Echinacoside thiab acteoside cov ntsiab lus ntawm CD hmoov tau txiav txim siab raws li peb txoj kev dhau los. (Zhao et al. 2011). Tom qab ntawd, txhua 100 g qhuav hmoov yog boiled nyob rau hauv 1.5 L dej rau 1.5 teev. Txias ntawm chav tsev kub, cov dej cawv sib tov tau centrifuged ntawm 2000 rpm rau 10 min (Eppendorf 5810R, Lub teb chaws Yelemees) thiab cov precipitate tau reextracted ob zaug nyob rau hauv tib yam mob. Cov supernatants sib xyaw ua ke tau tsom los ntawm lub tshuab nqus tsev rotaryevaporator (Heidolph labourota 4001-ntau, Lub teb chaws Yelemees) ntawm 60 degree, thiab tom qab ntawd lyophilized thiab yielded txog 42 feem pua ​​​​cov hmoov qhuav extract. Thaum ua qhov kev sim tom ntej, cov extract qhuav tau yaj cov dej huv hauv chav sov.

Kev sim tsiaj

Cov neeg laus txiv neej Sprague-Dawley nas (200-220g) tau txais los ntawm Kev Tshawb Fawb Tsiaj

Center of Guangdong Province (Guangzhou, PR Tuam Tshoj), tuav tswj qhov kub thiab txias nrog 12-hr teeb/darkcycles ntawm chav tsev kub (22-24 degree) thiab constanthumidity ({3}} feem pua) nyob rau hauv SPF (tshwj xeeb pathogen-free) kuaj thiab pub ad libitum. Cov txheej txheem sim tau ua raws li NationalInstitutes of Health Guide for Care and Use of LaboratoryAnimals.Experimental process.

Ib lub lis piam ua ntej kev tswj hwm, tsib caug txiv neej SD nas tau kho nrog sodium pentobarbital intraperitoneally (40 mg/kg) (Jung et al. 1999) thiab muab faib rau tsib pawg: tshwj tsis yog cov nas nyob hauv pawg neeg ua haujlwm sham (Sham), tus so yog ob tog castrated los ntawm txoj kev scrotal (Anderes 2003, Schlatt li al. 2002). Lub caij no, cov pab pawg CDtreated tau muab tshuaj intragastrically nrog 0.45g / kg (tsawg koob ntawm CD extract, LCD ntxiv rau Cas), 0.9g / kg (qhov nruab nrab koob ntawm CD extract, MCD ntxiv Cas) thiab 1.8g / kg (siab koob tshuaj CD. extract, HCD plus Cas) CD extract, raws li, thaum lub sham thiab castrated pab pawg neeg raug tswj xyuas nrog tus nqi sib npaug ntawm cov dej xwb.Lub cev qhov hnyav tau coj mus rau qhov pib ntawm cov txheej txheem thiab tom qab ntawd txhua lub lim tiam. Tag nrho cov nas tau ua laparotomy tam sim ntawd tom qab kev tshem tawm kev tuag thiab tom qab qhov hnyav ntawm cov kab mob hauv qab no: cov qog nqaij hlav, cov qog nqaij hlav, cov qog prostate thiab cov leeg nqaij.

Noov erectile teb ntsuas

Kev ua haujlwm erectile ntawm nas nyob rau hauv txhua pab pawg tau soj ntsuam raws li txoj kev dhau los (Ji li al. 2009, Luo et al.2006). Luv luv, lub stainless-steel bipolar electrode (BL420F Data Acquisition & Analysis System, TmeTechnology Co., Ltd, Chengdu, PR Tuam Tshoj) tau ua tib zoo muab tso rau ntawm noov kom txog thaum nws tau tsa nrog lub mem tes ntawm lub zog stimulation (5 V) 0.2 s. Lub sij hawm los ntawm stimulationto erection tau sau tseg raws li qhov noov erectile latency.Thaum lub noov erectile teb tau soj ntsuam, lub electrodecould yuav raug rho tawm ntawm noov, thiab lub sij hawm ntev lub noov khaws cia erection yog kaw raws li lub sij hawm erectile.

Kev kuaj ntshav thiab tshuaj ntsuam xyuas cov tshuaj hormones

Cov nas tsuag tau tshuaj loog nrog sodium pentobarbitalintraperitoneally (40mg / kg) (Jung et al. 1999).

Cov ntshav tau kos thiab txhaws ntawm 4 degree rau 4h, los ntawm cov ntshav tau sau los ntawm centrifugation ntawm 4000rpm rau 5 min ntawm 4 degree (Eppendorf 5810 R, Lub teb chaws Yelemees) thiab tam sim ntawd khaws cia ntawm -20 degree lub freezer kom txog thaum Kev soj ntsuam ntxiv.Tag nrho LH thiab FSH qib tau ntsuas los ntawm kev siv cov khoom siv hluav taws xob hauv xov tooj cua (RIA) (Beijing NorthInstitute of Biological Technology, Beijing, Tuam Tshoj) xa mus rau cov chaw tsim khoom cov lus qhia (Matsumoto etal. 1986). Raws li kev sib tw radioactive immunetechnique, I125-labeled LH (los yog FSH) tau siv nyob rau hauv qhov kev soj ntsuam.Liab, 100µl I125-labeled LH (los yog FSH) tus neeg sawv cev tau ntxiv rau 100µl cov ntshav cov qauv thiab incubated nrog 100µlpurified luav antibody ntawm 4 degree (los yog 37 degree) hmo ntuj. Tom qab ntawd, tus neeg sawv cev ntawm luav-tiv thaiv kab mob luav tau ntxiv rau hauv cov tshuaj tiv thaiv kab mob antigen thiab centrifuged (3500rpm, 15 min) kom cais tawm dawb thiab tshuaj tiv thaiv kab mob 125I-LH (lossis FSH). Tom qab ntawd, lub xov tooj cua ntawm precipitate tau ntsuas ntawm lub xov tooj cua tiv thaiv kab mob (DMF-96, Zhongcheng hluav taws xob thev naus laus zis thev naus laus zis Co., Ltd., Hefei, Tuam Tshoj). Cov kev txwv rhiab heev ntawm LH thiab FSHassay yog ob qho tib si 1.0mIU / ml.

STATISTICAL analysis

Kev tshuaj xyuas cov ntaub ntawv tau ua tiav siv SPSS version 16.0software.

Cov txiaj ntsig tau qhia raws li qhov txhais tau tias ± tus qauv sib txawv (SD). Cov ntaub ntawv Parametric tau txheeb xyuas siv kev tshuaj xyuas ib-txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li kev xeem tom qab hoc LSD. Cov ntaub ntawv nonparametric tau txheeb xyuas los ntawm Kruskal-Wallis test. Qhov sib txawv tau txiav txim siab qhov tseem ceeb yog tias qhov tshwm sim fordeviation tsawg dua 0.05.

TSEEM CEEB

Kev txiav txim kom muaj nuj nqis ntawm echinacoside andacetonide cov ntsiab lus ntawm CD hmoov

Cov ntsiab lus ntawm echinacoside thiab acetonide hauv CD hmoov yog 1.27±{2}} 01 feem pua ​​​​thiab 0.52±0.003 feem pua ​​, uas ua raws li qhov yuav tsum tau muaj ntawm Suav Pharmacopoeia. HPLCchromatogram ntawm CD hmoov tau pom hauv daim duab. 1.

(4.6mm × 250 mm, 5 µm, Waters, Milford, MA, USA);mob theem: 0.5 feem pua ​​(V/V) acetic acid aqueous solution(A) thiab methanol ( B) ingradient (0min, A: B=25:75; 10-30 min, A: B=40: 60; 30 min, A: B=0: 100); Tus nqi ntws: 1ml / min; Kev tshawb nrhiav wavelength: 330nm; kub: 30.

Daim duab 2: Noov erectile latency thiab ntev nyob rau hauv castratedrats. Cov ntaub ntawv raug qhia raws li qhov txhais tau tias ± SD (n=10). ##p<0.01 relative="" to="" sham-operated="" group,=""><0.01 relative="" to="" castrated="" group.="" abbreviation="" (below):="" sham,="" sham-operated="" group;="" cas,="" castration="" control="" group;="" lcd="" +cas="" (0.45g/kg),="" mcd="" +cas="" (0.9g/kg)="" and="" hcd="" +cas="" (1.8g/kg),="" three="" groups="" of="" cd="" extract="" treated="" in="" castrated="">

Fig. 3: Cov teebmeem ntawm CD extract ntawm cov ntshav LHconcentrations nyob rau hauv castrated nas. Cov ntaub ntawv raug qhia raws li qhov txhais tau tias ± SD (n=10). #p<0.05 relative="" to="" sham-operated="" group,="" *=""><0.05 relative="" to="" castrated="" group.="" body="" weights="" and="" relative="" accessory="" organs="" weights="" shown="" in="" table="" 1,="" all="" rats="" were="" of="" similar="" initial="" body="" weights="" with="" an="" increase="" from="" day="" 1="" to="" day="" 28.="" however,="" mean="" values="" in="" castrated="" group="" increased="" from="" 216.8±9.2="" g="" to="" 251.4±12.7,="" 283.3±15.1,="" 306.8±15.4="" and="" 322.2±18.4="" g,="" gaining="" less="" weights="" compared="" to="" the="" sham="" operated="" counterparts=""><0.05). there="" were="" no="" statistical="" difference="" between="" sham="" group="" and="" cd="" treated="">

Tsis tas li ntawd, qhov nruab nrab qhov hnyav ntawm lub caj pas preputial, qog prostate & seminal vesicle thiab levator ani leeg incastrated nas tau qis dua (p<0.05) than="" those="" of="" normal="" controls,="" respectively.="" compared="" with="" the="" rats="" castrated="" only,="" the="" relative="" values="" of="" these="" organs="" weight="" tended="" to="" be="" higher="" under="" the="" treatment="" of="" cd="" extract.="" erectile="" response="" assessment="" in="" contrast="" to="" sham="" group,="" castration="" made="" the="" rats="" spend="" more="" time="" gaining="" penis="" erection="" under="" electric="" stimulation=""><0.01). however,="" the="" erectile="" latency="" was="" shortened="" by="" cd="" extract,="" exhibiting="" a="" significant="" decrease="" by="" 52.7%="" in="" lcd="" +cas="" group=""><0.01), 73.5%="" in="" mcd="" +cas="" group=""><0.01) and="" 74.5%="" in="" hcd="" +cas="" group=""><0.01), respectively.="" furthermore,="" castration="" shortened="" the="" erectile="" duration="" with="" the="" mean="" value="" of="" 37.2±17.3="" s="" compared="" to="" sham="" operated="" group=""><0.01). however,="" the="" erectile="" duration="" in="" rats="" was="" significantly="" prolonged="" approximately="" by="" 2.6-fold="" in="" mcd="" +cas="" group=""><0.01) and="" 2.1-fold="" in="" hcd="" +cas="" group=""><0.01). it="" showed="" no="" strict="" dose-related="" correlation.="" (fig.="">

Qib ntawm cov tshuaj hormones

Kev soj ntsuam los ntawm RIA kev soj ntsuam, qhov nruab nrab cov ntshav concentration ntawm LH hauv cov nas castrated yog 2.50± 0.34mIU / ml, uas qhia tias muaj kev nce ntxiv piv rau kev tswj hwm sham (p<0.05). however,="" the="" value="" descended="" about="" 49.2%="" in="" mcd="" +cas="" group="" relative="" to="" that="" in="" castrated="" control=""><0.05) (fig.="" 3).="" shown="" in="" fig.="" 4,="" no="" statistical="" differences="" were="" observed="" among="" all="" groups="" on="" fsh="">

Kev sib tham

Peb cov txiaj ntsig tau pom tias castration ua rau lub sijhawm ntev ntawm qhov chaw mos erectile latency thiab luv luv theerectile duration, nrog rau cov khoom siv organsatrophy zoo kawg li. Txawm li cas los xij, CD extract tau yooj yim rau qhov noov erectileresponse thiab modulated cov ntshav LH qib mus rau qee qhov hauv cov nas tsuag.Qhov erection spontaneous yog txhawb nqa thiab tswj hwm los ntawm androgen ntawm cellular, molecular, andphysiologic mechanisms, nrog rau kev so ntawmcavernosal smooth 0, 0. Talha et al. 2007). Cov qib testosterone tsis txaus ua rau txo qis hauv kev so (McClure 1988), uas ua rau cov txiv neej impotence (Bivalacqua li al. 2000).

Tshwj xeeb hauv cov nas, nws tau tswj hwm los ntawm ntau qhov systemictestosterone ntau npaum li {{{0}}} feem pua ​​, hauv qab no cov lus teb theerectile yog attenuated nyob rau hauv ib koob tshuaj-dependentmanner (Armagan li al. 2006). Hauv kev tshawb fawb tam sim no, thetestosterone concentration hauv kev ncig yog tsawg dhau los ua kom tau raws li qhov kev xav tau txwv (0.02ng / ml) hauv txhua tus nas castrated (cov ntaub ntawv tsis tau pom hauv cov txiaj ntsig). Txawm li cas los xij, CDextract ua rau txo qhov noov erectile latency atthree dosages thiab ncua lub sij hawm erectile ntawm 0.90g / kg thiab 1.8 g / kg. Nws yog tej zaum qhov tshwm sim ntawm androgen-zoo li cov nyhuv ntawm CD tau hais hauv qee qhov kev tshawb fawb yav dhau los (Nws li al. 1996a, He et al. 1996b). Qee cov khoom siv hauv nruab nrog cev, xws li prostate thiab seminalvesicle, yog androgen nyob ntawm, yog li qhov hnyav hloov tuaj yeem cuam tshuam qhov Kev hloov pauv hauv endocrine xwm txheej (Trisomboon et al. 2007, Jang et al. 2011).

Nws tshwm sim theem pib ntawm testosterone, hauv qab no uas themetabolism ntawm cov kabmob no tau inhibited (Trachtenberg1985). Nws tau raug lees paub hauv peb txoj kev tshawb fawb tias castrationinduced cov khoom siv hauv cov menyuam yaus qhov hnyav kom txo qis vim qhov tsis muaj testosterone.Testosterone feem ntau yog zais los ntawm testis Leydig celland tswj los ntawm cov tshuaj hormones hypophysial, xws li cov tshuaj hormones (LH) thiab follicle-stimulatinghormone (FSH), uas ua si. lub luag haujlwm tseem ceeb hauv kev tswj xyuas qhov noov ua haujlwm ntawm cov tsiaj castrated (Robert li al. 2004). Raws li lub hypothalamus-pituitary-gonad (HPG) axis yog kaw-voj tawm tswv yim tswj mechanism, qis gonadal cov tshuaj hormones tuaj yeem tswj qhov tso tawm ntawm LH thiab FSH nyob rau hauv lem (Morales li al. 2004). Peb txoj kev tshawb fawb tau tshawb xyuas tias cov qib ntshav LH tau nce ntau hauv cov nas tsuag thiab tom qab ntawd nqis mus rau qib qis los ntawm CD extract ntawm 0.90 g / kg, uas tau pom zoo nrog cov lus xaus dhau los uas nce LHlevel tau cuam tshuam nrog kev pheej hmoo siab ntawm erectiledysfunction (Kupelian li al. 2006).

Nyob rau hauv testis, LHstimulates lub synthesis thiab tso tawm ntawm testosterone, cov lus teb rau kev tswj ntawm uas yog ib tug kuj zoo ib yam, thaum FSH ncaj qha ua rau cov Sertoli hlwb rau kev noj qab haus huv nrog thefeedback tswj complex (Hellqvist li al. 2008). Cov kev tshawb pom no tau muaj kev txhawb nqa ntau dua rau qhov kev xav tias CDexpressed androgen zoo li cov nyhuv, uas tej zaum yuav yog lub luag haujlwm tseem ceeb rau kev txo cov ntshav LH ntau dua li FSH.Nyob rau hauv kev soj ntsuam tam sim no, kev cuam tshuam ntawm kev loj hlob ntawm lub cev hnyav hauv cov txiv neej nas yog zoo ib yam li cov kev tshawb pom hauv yav dhau los (Christoffersen et al. 2006), hos contrary to the results in pojniam nas (Pantaleão etal. 2010). CD extract tsis tuaj yeem txhim kho lub cev qhov hnyav, uas yog tej zaum vim qhov kev kho mob tsis txaus lossis qhov cuam tshuam rau kev noj qab haus huv tag nrho.Nyob rau hauv xaus, castration prolonged noov erectilelatency thiab shortened lub sij hawm tseem ceeb, withserum LH nce thiab accessory kev sib deev kab mob atrophy Txawm li cas los, CD extract tau yooj yim rau qhov noov erectileresponse los ntawm shortening lub erectile latency thiab prolonging lub erectile lub sij hawm nyob rau hauv castrated nas. Cov teebmeem no tau tshwm sim los ntawm cov nyhuv zoo li androgen, uas tau txais kev txhawb los ntawm kev soj ntsuam tias CD extract normalized theserum LH concentrations tshwj xeeb tshaj yog nyob rau ntawm qhov ntau npaum ntawm 0.90g / kg.

Txawm li cas los xij, cov pov thawj ntxiv yog xav tau los ua kom pom cov txiaj ntsig ntawm CD extract tiv thaiv ED athypothalamic thiab limbic qib thiab ntau qhov kev saib xyuas tuaj yeem tau txais rau cov tshuaj tseem ceeb hauv ntiaj teb niaj hnub no.