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

Li Gu, Wen Ting Xiong, Yan Lei Zhuang, Jian Shuang Zhang, thiab Xin Liu*

Khoom noj khoom haus thiab kev noj qab haus huv Engineering Research Center ntawm Lub Xeev Education Ministry, Sun Yat-sen University, Guangzhou, Suav

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 ntawm noov erectile teb tau tshawb xyuas nyob rau hauv txoj kev tshawb 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 erectile teb tau ntsuas thiab cov tshuaj hormones hauv cov ntshav tau soj ntsuam thaum kawg ntawm qhov kev sim. Nws tau raug soj ntsuam tias qhov erectile latency tau ntev dua thiab lub sijhawm erectile luv luv luv hauv cov nas castrated piv rau kev tswj hwm sham. Txawm li cas los xij, CD extract txo qhov erectile latency thiab ncua lub sijhawm erectile kom txo qis qhov tsis zoo ntawm castration. Ntawm qhov ntau npaum ntawm 0.9g / kg, CD extract tswj cov ntshav luteinizing hormone concentration mus rau theem ib txwm nyob hauv cov nas tsuag. Cov kev tshawb pom no tau qhia tias CD pab txhawb qhov noov erectile teb thiab modulated cov ntshav cov tshuaj hormones rau qee yam.

Nyem cistanche tonic khoom

Taw qhia

Cistanche Hoffmg. Et Link, ib tug ntawm cov genera ntawm Orobanchaceae tsev neeg, feem ntau yog faib nyob rau hauv arid av thiab suab puam nyob rau hauv sab qaum teb hemisphere, xws li Inner Mongolia, Xinjiang, thiab Ningxia autonomous cheeb tsam ntawm Tuam Tshoj, Iran, India, Mongolia, thiab lwm yam. Hom kab no yog cov tshuaj tua kab mob perennial, uas feem ntau txuas rau cov hauv paus hniav ntawm cov xuab zeb kho cov nroj tsuag xws li Haloxylox ammo dendron, H. persicum, Kalidium foliated, thiab Tamarix nroj tsuag, thiab lwm yam. (Jiang and Tu 2009).

Herba Cistanche, qia ntawm Cistanche hom, tau raug suav hais tias yog ib qho zoo tshaj tonic thiab tau txais kev qhuas 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) yog qhov tsis txaus uas tau sau los ua ib qho ntawm cov nroj tsuag Class II xav tau kev tiv thaiv hauv Suav teb, yog li lwm hom ntawm cov genus no, xws li C. salsa thiab C. sinensis, kuj yog siv los hloov hauv qee qhov chaw. Los ntawm thawj zaug sau tseg hauv Shen Nong's Suav Cov Khoom Siv Medica (ib qho khoom siv tshuaj thaum ub), CD tau xav kom ua kom lub raum ua kom lub raum thiab ua kom lub raum "Yang" hauv kev kho mob raws li kev cai suav tshuaj suav tshuaj, uas tau liam tias muaj txiaj ntsig zoo rau kev loj hlob, kev loj hlob, thiab fertility muaj nuj nqi. Lub caij no, cov CD kuj tau muab cov nqaij nyug, qos yaj ywm, los yog mov los ua cov zaub mov tonic los kho kev puas tsuaj ntau dhau 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 anecdotal no feem ntau muaj nyob hauv Suav incunabula, nrog cov ntaub ntawv tshawb fawb tsawg tau tham txog cov ntsiab lus niaj hnub pharmacological.

Erectile dysfunction (ED) yog ib yam mob uas tshwm sim los ntawm qhov tsis txaus erection ntawm noov lossis erection uas tsis kav ntev txaus rau cov txiv neej laus (Cohan thiab Korenman 2000). Nyob rau hauv cov lus teb rau niaj hnub, sophisticated, chemical agents, muaj ib tug tshiab kev txaus siab mus nrhiav tau cov tshuaj zoo thiab nyab xeeb los ntawm cov nroj tsuag rau kev kho mob ntawm ED (Adimoelja 2000). Ib txoj kev tshawb fawb yav dhau los (Liu li al. 2009) tau qhia tias CD electuary accelerated 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 cov tsiaj, kev phais castration nrog ob sab orchiectomy tau raug suav tias yog tus qauv ntawm castration rau kev tshawb xyuas ED (Tombal 2005). Yog li, txoj 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 erectile latency thiab lub sijhawm ntev ntawm cov nas castrated..

Cov khoom siv thiab cov txheej txheem

Tshuaj thiab khoom siv

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 tau yuav los ntawm Sigma-Aldrich (St. Louis, MO, USA). Cov dej huv huv tau tsim los siv Millipore Milli-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, Tuam Tshoj) thiab hauv av rau hauv cov hmoov zoo nrog lub pulverizer dhau los ntawm 60 mesh sieve.

Kev tshuaj xyuas HPLC thiab kev npaj cov khoom cog

Echinacoside thiab acteoside cov ntsiab lus ntawm CD hmoov tau txiav txim siab raws li peb txoj kev dhau los (Zhao li al. 2011). Tom qab ntawd, txhua 100 g ntawm qhuav hmoov yog boiled nyob rau hauv 1.5

l dej rau 1.5 teev. Txias ntawm chav tsev kub, cov dej cawv sib xyaw 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 ua ke tau concentrated los ntawm lub tshuab nqus tsev evaporator (Heidolph laborota 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 cov kev sim tsiaj tom ntej, cov extract qhuav tau yaj hauv dej huv ntawm chav tsev kub.

Kev sim tsiaj

Cov neeg laus txiv neej Sprague-Dawley nas (200-220g) tau txais los ntawm Lub Chaw Kho Mob Kev Tshawb Fawb Tsiaj ntawm Guangdong Xeev (Guangzhou, PR Tuam Tshoj), khaws cia nyob rau hauv qhov ntsuas kub nrog 12-hr teeb / tsaus voj voog ntawm chav ntsuas kub (22-24 degree ) thiab cov av tsis tu ncua (40-70 feem pua ​​) hauv SPF (tsis muaj kab mob tshwj xeeb) kuaj thiab pub ad libitum. Cov txheej txheem sim ua raws li National Institutes of Health Guide for Care and Use of Laboratory Tsiaj.

Kev sim txheej txheem

Ib lub lis piam ua ntej kev tswj hwm, tsib caug txiv neej SD nas tau muab tshuaj loog nrog sodium pentobarbital intraperitoneally (40 mg/kg) (Jung et al. 1999) thiab muab faib rau tsib pawg: tshwj tsis yog nas nyob rau hauv pawg neeg ua haujlwm sham (Sham ), tus so yog ob tog castrated los ntawm scrotal mus kom ze (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 ntawm CD extract, HCD plus Cas) CD extract, ntsig txog, thaum lub sham thiab castrated pawg tau tswj hwm tus nqi sib xws ntawm cov dej xwb. Kev ntsuas lub cev hnyav tau muab coj los ua thaum pib ntawm cov txheej txheem thiab tom qab ntawd txhua lub lim tiam. Txhua tus nas tau ua laparotomy tam sim ntawd tom qab tuag rau kev tshem tawm thiab tom qab qhov hnyav ntawm cov kab mob hauv qab no: cov qog ua ntej, cov qog nqaij hlav, cov qog prostate, thiab levator ani leeg.

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, Tme Technology Co., Ltd, Chengdu, PR China) tau ua tib zoo muab tso rau ntawm noov kom txog thaum nws tau tsim nrog lub mem tes ntawm lub zog stimulation (5 V) 0.2 s. Lub sij hawm los ntawm stimulation mus rau erection tau kaw raws li noov erectile latency. Thaum pom qhov noov erectile teb, cov electrode yuav raug rho tawm ntawm noov, thiab lub sij hawm ntev uas noov khaws cia erection raug 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 pentobarbital intraperitoneally (40mg/kg) (Jung et al. 1999). Cov ntshav kuaj tau kos thiab txhaws ntawm 4 degree rau 4h, los ntawm cov ntshav tau sau los ntawm centrifugation ntawm 4000 rpm 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 rau thaum kev soj ntsuam ntxiv. . Tag nrho LH thiab FSH qib tau ntsuas los ntawm kev siv cov khoom siv hluav taws xob ua lag luam (RIA) (Beijing North Institute of Biological Technology, Beijing, Tuam Tshoj) xa mus rau cov chaw tsim khoom cov lus qhia (Matsumoto li al. 1986). Raws li kev sib tw xov tooj cua tiv thaiv kab mob, kuv125- sau npe LH (los yog FSH) tau siv rau hauv qhov kev xeem. Luv luv, 100µl I{12}}labeled LH (los yog FSH) tus neeg sawv cev tau ntxiv rau hauv 100µl cov qauv ntshav thiab incubated nrog 100µl purified luav antibody ntawm 4 degree (los yog 37 degree) thaum hmo ntuj. Tom qab ntawd, tus nees luav-tiv thaiv kab mob sib cais tau ntxiv rau cov tshuaj tiv thaiv kab mob antigen thiab centrifuged (3500 rpm, 15 min) kom cais cov dawb thiab cov tshuaj tiv thaiv kab mob 125I-LH (lossis FSH). Tom qab ntawd, lub xov tooj cua ntawm cov nag lossis daus 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, Suav). Cov kev txwv rhiab heev ntawm LH thiab FSH kev soj ntsuam yog ob qho tib si 1.0mIU / ml.

STATISTICAL analysis

Kev tshuaj xyuas cov ntaub ntawv tau ua tiav siv SPSS version 16.0 software. Cov txiaj ntsig tau qhia raws li qhov txhais tau tias ± tus qauv sib txawv (SD). Cov ntaub ntawv parametric tau raug tshuaj xyuas siv kev tshuaj ntsuam 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 suav tias yog qhov tseem ceeb yog tias qhov tshwm sim ntawm qhov sib txawv tsawg dua 0.05.

TSEEM CEEB

Kev txiav txim siab ntawm echinacoside thiab acteoside cov ntsiab lus ntawm CD hmoov

Cov ntsiab lus ntawm echinacoside thiab acteoside 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. HPLC chromatogram ntawm CD hmoov tau pom hauv daim duab. 1.

Fig. 1

HPLC chromatographic profile ntawm Cistanche deserticola hmoov. Peak 1, echinacoside; Peak 2, acteoside. X axis, tuav lub sijhawm; Y axis, absorbance unit. HPLC tej yam kev mob: Dej HPLC system (Binary HPLC Pump1525, Refractive Index Detector 2414, Photodiode Array Detector 2996); nyob ruaj khov theem: Symmetry C18(4.6mm × 25{{10}} mm, 5 µm, Dej, Milford, MA, USA); mobile theem: 0.5 feem pua ​​(V/V) acetic acid aqueous tov (A) thiab methanol (B) muaj (0min, A: B=25:75; 10- 30 min, A: B{{16 }}: 60; 30 min, A: B=0: 100); Tus nqi ntws: 1ml / min; Kev tshawb nrhiav wavelength: 330nm; kub: 30.

Fig.2

Noov erectile latency thiab ntev nyob rau hauv castrated nas. Cov ntaub ntawv raug qhia raws li qhov txhais tau tias ± SD (n=10). ##p<0.01 relative to sham-operated group, **p<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 rats

Fig.3

Cov teebmeem ntawm CD extract ntawm cov ntshav LH concentrations 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, * p<0.05 relative to castrated group.

Lub cev hnyav thiab txheeb ze cov khoom siv hauv nruab nrog cev hnyav

Raws li pom nyob rau hauv Table 1, tag nrho cov nas muaj qhov sib npaug ntawm lub cev hnyav nrog qhov nce ntawm hnub 1 txog rau hnub 28. Txawm li cas los xij, qhov tseem ceeb hauv pawg castrated tau nce los ntawm 216.8 ± 9.2 g rau 251.4 ± 12.7, 283.3 ± 15.1, 306.8 ± 15.4. thiab 322.2 ± 18.4 g, nce qhov hnyav dua piv rau cov neeg ua haujlwm sham (p<0.05). There were no statistical differences between the sham group and CD treated group.

Fig.4

Cov teebmeem ntawm CD extract ntawm cov ntshav FSH concentrations hauv cov nas castrated. Cov ntaub ntawv raug qhia raws li qhov nruab nrab ± SD (n=10).

Cov theem ntawm cov tshuaj hormones

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

Kev sib tham

Peb cov txiaj ntsig tau pom tias castration ua rau lub sijhawm ntev ntawm qhov chaw mos erectile latency thiab txo lub sijhawm erectile, nrog rau cov khoom siv hauv nruab nrog cev zoo kawg nkaus atrophy. Txawm li cas los xij, CD extract tau yooj yim rau qhov noov erectile teb thiab hloov kho cov ntshav LH qib rau qee qhov hauv cov nas castrated.

Lub cev erection feem ntau yog txhawb nqa thiab tswj hwm los ntawm androgen ntawm cellular, molecular, thiab physiologic mechanisms, nrog rau kev so ntawm cov leeg nqaij (Traish li al. 2007, Talha li al. 2007) . Cov qib testosterone tsis txaus ua rau txo qis hauv nws qhov kev so (McClure 1988), uas ua rau cov txiv neej impotence (Bivalacqua li al. 2000). Tshwj xeeb tshaj yog nyob rau hauv nas, nws yog tswj los ntawm ib tug ntau yam ntawm systemic testosterone concentrations li 10-12 feem pua, hauv qab no uas cov erectile teb yog attenuated nyob rau hauv ib tug npaum li cas ntawm koob tshuaj (Armagan li al. 2006). Hauv kev tshawb fawb tam sim no, testosterone concentration hauv kev ncig yog tsawg dhau los ua kom tau raws li qhov kev xav tau (0.02ng / ml) hauv txhua tus nas castrated (cov ntaub ntawv tsis tau qhia hauv cov txiaj ntsig). Txawm li cas los xij, CD extract tau txo qis qhov noov erectile latency ntawm peb qhov ntau npaum thiab ua kom lub sijhawm erectile ntawm 0.90 g / kg thiab 1.8 g / kg. Nws yog tej zaum qhov tshwm sim ntawm cov nyhuv androgen zoo li CD hais hauv qee qhov kev tshawb fawb yav dhau los (He et al. 1996a, He et al. 1996b).

Qee cov khoom siv hauv nruab nrog cev, xws li prostate thiab seminal vesicle, yog androgen-dependant, yog li qhov kev hloov pauv hnyav tuaj yeem cuam tshuam txog kev hloov pauv hauv endocrine xwm txheej (Trisomboon li al. 2007, Jang et al. 2011). Nws tshwm sim ib theem pib ntawm testosterone, hauv qab no uas cov metabolism hauv cov kabmob no tau inhibited (Trachtenberg 1985). Nws tau raug lees paub hauv peb txoj kev tshawb fawb tias castration induced accessory reproductive organ 'weight to reduce vim testosterone deficiency.

Testosterone feem ntau yog secreted los ntawm testis Leydig cell thiab tswj los ntawm hypophysial cov tshuaj hormones, xws li luteinizing hormone (LH) thiab follicle-stimulating hormone (FSH), uas ua lub luag haujlwm tseem ceeb hauv kev tswj xyuas qhov chaw mos ntawm cov tsiaj castrated (Robert li al. 2{ {6}}04). Raws li lub hypothalamus-pituitary-gonad (HPG) axis yog kaw-voj tawm tswv yim tswj mechanism, tsawg 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 siab hauv cov nas castrated thiab tom qab ntawd nqis mus rau qib ib txwm los ntawm CD extract ntawm 0.90 g / kg, uas tau pom zoo nrog cov lus xaus dhau los uas nce qib LH tau cuam tshuam nrog kev pheej hmoo siab ntawm erectile kawg. (Kupelian et al. 2006). Nyob rau hauv testis, LH stimulates lub synthesis thiab tso tawm ntawm testosterone, lub tswv yim tswj ntawm uas yog kuj zoo ib yam, thaum FSH ncaj qha ua nyob rau hauv Sertoli hlwb rau kev noj haus nrog lub tswv yim tswj complex (Hellqvist li al. 2008). Cov kev tshawb pom no tau muaj kev txhawb nqa ntau dua rau qhov kev cia siab tias CD qhia txog cov nyhuv zoo li androgen, uas yuav yog lub luag haujlwm tseem ceeb rau kev txo cov ntshav LH ntau dua li FSH.

Hauv kev soj ntsuam tam sim no, kev cuam tshuam ntawm kev loj hlob ntawm lub cev hnyav hauv cov txiv neej nas zoo ib yam li qhov kev tshawb pom hauv kev tshawb fawb yav dhau los (Christoffersen li al. 2006), tsis zoo li cov txiaj ntsig ntawm poj niam nas (Pantaleão et al. 2010). CD extract tsis tuaj yeem txhim kho lub cev hnyav, uas tej zaum yog vim lub sijhawm kho tsis txaus lossis cuam tshuam rau tag nrho cov kev noj qab haus huv.

Hauv kev xaus, castration prolonged noov erectile latency thiab shortened lub sij hawm tseem ceeb, nrog rau cov ntshav LH nce thiab accessory txiv neej pw atrophy.

Txawm li cas los xij, CD extract tau yooj yim rau qhov noov erectile teb los ntawm shortening lub erectile latency thiab prolonging lub sij hawm erectile nyob rau hauv castrated nas. Cov teebmeem no tej zaum yog vim cov nyhuv zoo li androgen, uas tau txais kev txhawb nqa los ntawm kev soj ntsuam tias CD extract normalize cov ntshav LH concentrations, tshwj xeeb tshaj yog nyob rau ntawm ntau npaum li cas ntawm 0.90 g / 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 ntawm qib hypothalamic thiab limbic thiab yuav tau txais kev saib xyuas ntau dua rau cov tshuaj tseem ceeb hauv lub ntiaj teb niaj hnub no.

TXOJ CAI

Txoj haujlwm no tau txais kev txhawb nqa los ntawm cov nyiaj pab los ntawm Science thiab Technology Support los ntawm Project hauv Xinjiang Uygur Autonomous Region (PR Tuam Tshoj) (200840102-15).

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