Qhov sib txawv ntawm Cov Khoom Siv Thiab Kev Siv Tshuaj Kho Mob Ntawm Cov Khoom Raw Thiab Cov Khoom Siv Caw Ua Haujlwm Los Ntawm Cistanches Herba

Abstract:

Lub hom phiajTxhawm rau sib piv cov kev hloov pauv ntawm cov tshuaj muaj pes tsawg leeg thiab kev siv tshuaj kho mob ntawm Roucongrong (Cistanches Herba) ua ntej thiab tom qab ua tiav, txhawm rau qhia meej txog lub hauv paus ntsiab lus ntawm stewing nrog cawv.

Cov txheej txheem LC-MS tau siv los txheeb xyuas cov tshuaj sib xyaw ua ntej thiab tom qab kev sib xyaw ntawm Cistanches Herba, ua ke nrog ntau yam kev txheeb xyuas txheeb xyuas kom pom qhov tsis sib xws ntawm cov khoom ua ntej thiab tom qab stewing nrog cawv. Qhov kev ua tau zoo sib txawv tau ua pov thawj los ntawm cov qauv nas ntawm hydrogenated cortisonekidney yang deficiency.

Cov txiaj ntsigTag nrho ntawm 45 chemical Cheebtsam tau txheeb xyuas los ntawmCistanches Herba cov qauv; Cov khoom sib xyaw ua ke rau qhov sib txawv ntawmCistanches Herba thiab wine-stewed Cistanches Herbayogechinacoside cov tshuaj, verbascoside, isoacteoside thiab tubuloside B; Tag nrho cov phenylethanoid glycosides extract los ntawm Cistanches Herba muaj cov txiaj ntsig zoo ntawm kev txhim kho cov qauv nas nrog rau lub raum yang deficiency tshaj cawv-stewed Cistanches Herba. Xaus Cov ntsiab lus ntawm Cistanche hloov tom qab raug muab tso rau hauv cawv, thiab cov nyhuv ntawm lub raum yang deficiency tau txhim kho. Cov khoom xyaw ntawm kev hloov pauv ntawm Cistanches Herbato wine-stewed Cistanches Herba tej zaum yuav yog cov khoom xyaw nquag uas txhawb nqa lub raum thiab txhawb nqa yang.

Cov lus tseem ceeb: Cistanches Herba; wine-stewed Cistanches Herba;lub raum yang deficiency;echinacoside cov tshuaj;lus verbascoside; isoacteoside; tubuloside B

HIGH-QUALITY CISTANCHE tshuaj ntsuab muag

Txhawb Kev Pabcuam Ntawm Wecistanche

Email: wallence.suen@wecistanche.com

Whatsapp / Tel: +86 15292862950

Nws yog lub npe hu ua "ginseng" thiab feem ntau yog faib nyob rau hauv Inner Mongolia, Ningxia (Azuqi), Gansu (Changma), Xinjiang thiab lwm cheeb tsam [1]. Cistanche yog sov nyob rau hauv qhov, qab zib thiab qab ntsev nyob rau hauv saj, thiab nkag mus rau lub raum thiab cov hnyuv loj meridians. Nws muaj cov txiaj ntsig ntawm kev txhawb nqa lub siab thiab lub raum, muaj txiaj ntsig zoo rau cov ntsiab lus thiab cov ntshav, thiab moistening cov hnyuv thiab txo cov cem quav. Nws tuaj yeem siv rau lub raum yang deficiency, impotence thiab infertility, mob ntawm lub duav thiab lub hauv caug, tsis muaj zog ntawm cov leeg thiab pob txha, plab hnyuv qhuav thiab cem quav [2]. Kev tshawb fawb niaj hnub no qhia tau hais tias lub cev nqaij daim tawv nrog cov nplai nplooj ntawm Cistanche feem ntau muaj ntau yam tshuaj lom neeg xws li phenylethanol glycosides, cyclopentane ether terpenes thiab lawv cov glycosides, lignans, qab zib, thiab lwm yam. [3], thiab muaj ntau yam tshuaj xws lianti-aging,antioxidant,anti-dementia,tiv thaiv qaug zog, thiab moistening cov hnyuv thiab tshem tawm cem quav [4].

Tom qab stewed nrog cawv, lub raum-tonifying thiab yang-txhim kho nyhuv ntawm Cistanche yog txhim kho. Lub hauv paus ntsiab lus ntawm kev ua haujlwm tsis tau piav qhia meej. Fan Yanan et al. [5-7] tau kawm txog qhov cuam tshuam ntawm Cistanche ntawm nas defecation, kev tiv thaiv kab mob, enteric neurotransmitters thiab lwm yam gene nthuav qhia ua ntej thiab tom qab ua. Da et al. siv cov cawv ntawm phenylethanol glycosides ua ntej thiab tom qab ua tiav los piav qhia nws txoj hauv kev ua haujlwm; Zhang Chaw et al. [8] siv cov kev hloov pauv hauv cov ntsiab lus ntawm phenylethanol glycosides ua ntej thiab tom qab ua tiav los piav qhia txog cov txheej txheem ua haujlwm; Zhang Shuyun et al. [9] siv cov kev hloov pauv hauv cov ntsiab lus ntawm betaine ua ntej thiab tom qab ua tiav los piav qhia cov txiaj ntsig ntawm kev ua haujlwm ntawm Cistanche. Cov ntaub ntawv uas twb muaj lawm ntawm kev ua ntawm Cistanche feem ntau yog tsom rau cov tshuaj pharmacology, thiab kev tshawb fawb ntawm cov khoom siv tseem tsom rau kev hloov pauv ntawm cov ntsiab lus ntawm cov khoom tshwj xeeb, tsis muaj kev tshawb fawb txog kev sib txawv ntawm cov khoom sib txawv. Qhov kev sim no siv cov kua chromatography-mass spectrometry (LC-MS) thev naus laus zis ua ke nrog ntau qhov kev txheeb xyuas txheeb cais los txheeb xyuas cov tshuaj lom neeg hauv Cistanche, txhawm rau nrhiav qhov sib txawv ntawm cov khoom nyoos Cistanche thiab wine-stewed khoom, thiab kawm qhov txawv ntawm qhov ua tau zoo ntawm raw Cistanche thiab wine-stewed khoom los ntawm tus qauv nas ntawm lub raum yang deficiency induced los ntawm hydrocortisone, thiaj li piav qhia cov khoom hauv paus ntawmCistanche hauv tonifying lub raum thiab txhawb yangthiab cov txheej txheem ua ntawm wine-stewed Cistanche.

HIGH-QUALITY CISTANCHE tshuaj ntsuab muag

1 Cov khoom siv

1.1 Cov cuab yeej

Qhov kawg 3000 UPLC-LTQ Orbitrap XL electrostatic teb tandem ion ntxiab high-resolution mass spectrometer (Thermo Fisher Scientific, USA), Theraio Xcaliber workstation; BT125D 1/100000 hluav taws xob tshuav nyiaj li cas (Sartorius Instrument System, Lub teb chaws Yelemees); KQ500DE CNC ultrasonic nqus tsev vacuum (Kunshan Ultrasonic Instrument Co., Ltd.); XHF-D high-speed disperser (Guangzhou Hu Ruiming Instrument Co., Ltd.); Multiskan FC microplate nyeem ntawv (Thermo Scientific); HC-3518 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); BW-YLS-1B multifunctional nas voluntary act recorder (Shanghai Ruanlong Technology Development Co., Ltd.).

1.2 Tshuaj

Mass spectrometry qib methanol (Fisher); pawg spectrometry qib formic acid (mass feem 99%, Sigma Aldrich); dej (Watsons distilled dej); Jinkui Shenqi Pills (Beijing Tongrentang Technology Development Co., Ltd. Pharmaceutical Factory, batch naj npawb 1701653); hydrocortisone succinate sodium ntsev (Tianjin Biochemical Pharmaceutical Co., Ltd., batch tooj 011705067); superoxide dismutase (SOD) cov khoom siv (batch naj npawb 20170115) thiab malondialdehyde (MDA) cov khoom siv (batch tooj 20170117) tau yuav los ntawm Nanjing Jiancheng Biotechnology Co., Ltd.; Testosterone (T) thiab estradiol (E2) tau txiav txim siab ntawm Qingdao Kechuang Quality Inspection Center, thiab creatinine (CREA) thiab ntshav urea nitrogen (BUN) tau txiav txim siab ntawm Lub Chaw Ntsuam Xyuas thiab Kev Ntsuas ntawm Inner Mongolia Medical University.

1.3 Tsiaj

100 txiv neej SD nas, hnyav 180-200 g, tau yuav los ntawm Xinglong Experimental Animal Farm hauv Haidian District, Beijing, daim ntawv tso cai naj npawb: SCXK (Beijing) 2016-0003. Qhov kev sim tsiaj no tau pom zoo los ntawm Pawg Saib Xyuas Kev Ncaj Ncees ntawm Beijing University of Chinese Medicine, tus lej coj ncaj ncees: BUCM-4-2022091301-3095.

1.4 Cov qauv

Rau batch ntawm Cistanche deserticola tau txheeb xyuas qhov tseeb los ntawm xibfwb Li Xiangri ntawm Lub Tsev Kawm Ntawv ntawm Suav Tshuaj, Beijing University of Suav Tshuaj, thiab tau muab los ntawm cov tawv nqaij qhuav nrog nplai nplooj ntawm Cistanche deserticola YC Ma, ib tsob nroj ntawm tsev neeg Orobanchaceae. Cov ncauj lus kom ntxaws ntawm cov qauv yog qhia hauv Table 1.

Table 1 Cov ntawv qhia ntawm Cistanches Herba cov qauv

2 Txoj kev

2.1 Kev npaj ua qauv

2.1.1 Kev npaj cov khoom nyoos Siv Cistanche deserticola huv si, muag nws, txiav rau hauv tuab slices, thiab qhuav nws.

2.1.2 Kev npaj cov khoom lag luam wine-stewed

Noj 100 g ntawm Cistanche deserticola slices, ntxiv 30 mL ntawm mov wine, sib tov zoo, thiab stew kom txog thaum lub cawu cub absorbed. Stew nws hauv dej rau 12 teev, tshem tawm thiab cia nws txias me ntsis, thiab qhuav nws. Saib daim duab 1 rau Cistanche deserticola nyoos thiab cawv-stewed Cistanche deserticola cov qauv.

2.2 Kawm txog qhov sib txawv ntawm cov khoom ntawm Cistanche deserticola ua ntej thiab tom qab stewing hauv cawv

2.2.1 Kev npaj kev daws teeb meem

Noj li ntawm {{0}}.2 g ntawm cov qauv hmoov qhuav (dhau los ntawm 65-mesh sieve), hnyav kom raug, tso rau hauv 50 mL xim av volumetric hwj, ntxiv 25 mL ntawm 50% methanol aqueous tov, kaw nruj nreem, hnyav qhov hnyav, tsau rau 30 feeb, ultrasonically kho rau 40 feeb (lub hwj chim 250 W, zaus 35 kHz), txias, hnyav dua, ntxiv 50% methanol aqueous tov los ua kom poob loj, co Zoo, cia sawv ntsug, lim cov supernatant los ntawm 0.22 μm microporous lim membrane, thiab tso cov filtrate rau hauv lub raj mis xim av theem kom tau txais cov tshuaj.

HIGH-QUALITY CISTANCHE tshuaj ntsuab muag

2.2.2 Chromatographic tej yam kev mob

ACQUITY UPLCTM BEH C18 chromatographic kem (100 mm × 2.1 mm, 1.7 μm), kem kub ntawm 35 degree; qauv chamber kub ntawm 10 degree, qauv txhaj tshuaj ntim ntawm 3 μL, methanol (A) -0.1% formic acid aqueous tov (B) raws li theem mobile, gradient elution: 0-4 min, 10%{ {13}}% methanol; 4-18 min, 26% -40% methanol; 18-24 min, 40% -100% methanol; 24-28 min, 100% methanol. Lub ntim ntws tus nqi yog 0.3 mL / min, thiab qhov ntsuas wavelength yog 330 nm.

2.2.3 Mass spectrometry tej yam kev mob

Electrospray ionization qhov chaw (ESI) tau siv, tsis zoo ion hom scanning, capillary kub yog 350 degree, sheath gas ntim tus nqi yog 30 arb, auxiliary gas volume flow rate yog 10 arb, spray voltage yog 3

KV, capillary voltage yog −35 V, raj lens voltage yog −110 V, scanning range yog m/z 100-1 000, data collection system yog Xcalibur 2.1.

2.2.4 Kev txheeb xyuas cov khoom tseem ceeb (PCA)

Tom qab tau txais ob-dimensional matrix ntawm cov ntaub ntawv khaws cia lub sij hawm muaj tseeb, cov cuab yeej vaj huam sib luag yuav tso tawm tag nrho cov ntaub ntawv khaws cia kom raug, thiab PCA txoj kev ntawm cov kev txheeb cais txuas ntxiv yuav raug siv los tshuaj xyuas.

2.2.5 Orthogonal ib feem tsawg squares cais cais (OPLS-DA) LCMS muab cov kev sib cais siab ntawm cov kua chromatography nrog lub siab rhiab heev ntawm huab hwm coj spectrometry. Cov ntaub ntawv khaws tseg yog qhov loj heev, thiab ntau qhov kev txheeb xyuas txheeb cais yuav tsum tau ua cov ntaub ntawv. OPLS-DA muab cov ntaub ntawv faib ua ob pawg thiab siv cov qauv kev soj ntsuam xyuas kom tau txais ntau tus qauv orthogonalized, uas qhia meej qhov sib txawv ntawm ob pawg ntawm cov qauv. Nyob rau hauv daim duab ntxiv S-Plot hom ntawv, txhua lub ntsiab lus sawv cev rau lub sijhawm khaws cia cov ntaub ntawv muaj tseeb, X-axis sawv cev rau qhov sib txawv, qhov deb ntawm cov ntaub ntawv taw tes yog los ntawm xoom point, ntau dua qhov kev koom tes ntawm qhov taw tes rau qhov piv txwv sib txawv. ; Y-axis sawv cev rau kev sib raug zoo ntawm cov qauv hauv tib pawg qauv, qhov deb dua qhov tseeb qhov loj thiab tuav lub sij hawm cov ntaub ntawv khub yog los ntawm xoom point, qhov kev sib raug zoo dua. Yog li ntawd, cov ntaub ntawv ceev ceev ceev ntawm ob qhov kawg ntawm S-shaped nkhaus sawv cev rau cov yam ntxwv zoo tshaj plaws ions los ntawm txhua pawg qauv.

HIGH-QUALITY CISTANCHE tshuaj ntsuab muag

2.2.6 Mass spectrometry qauv tsom xam

Tom qab tau txais cov yam ntxwv ions, lub ntsiab lus muaj pes tsawg leeg thiab kev sib xyaw ua ke tau ua los ntawm kev suav cov txheeb ze txheeb ze molecular loj thiab sib txuas cov ntaub ntawv uas twb muaj lawm thiab cov ntaub ntawv cuam tshuam los txheeb xyuas cov khoom sib txawv.

2.3 Kawm txog qhov sib txawv ntawm kev ua tau zoo ua ntej thiab tom qab stewing nrog cawv

2.3.1 Kev npaj ntawm Cistanche deserticola extract

Cistanche deserticola (batch naj npawb 160901) thiab coj cov cawv txiv hmab-stewed Cistanche deserticola hmoov tau muab rho tawm raws li cov xwm txheej hauv qab "2.2.1", lim tom qab kev kho ultrasonic, rov ua ob zaug, cov lim tau ua ke, concentrated los ntawm lub tshuab nqus tsev distillation, thiab khov- qhuav.

2.3.2 Tsiaj sim pab pawg thiab tswj tshuaj

Fig. 1 Cistanches Herba A thiab wine-stewed Cistanches Herba B

(1) Kev Pabcuam: Cov nas tau txais kev nkag mus rau dej thiab noj zaub mov dawb. Tom qab 1 lub lim tiam ntawm kev yoo mov, lawv tau muab faib ua 5 pawg (Table 2), nrog rau 10 nas nyob rau hauv txhua pab pawg, uas yog ib pab pawg, pawg qauv, cov Cistanche deserticola nyoos pawg (PR), wine-stewed Cistanche deserticola pawg (PW) , thiab tswj zoo Jinkui Shenqi Pill pab pawg (JK). (2) Kev ua qauv: Tsuas yog cov pab pawg ib txwm muaj, plaub pawg lwm pawg tau txhaj tshuaj hydrocortisone sodium succinate im, nrog rau koob tshuaj 20 mg / kg, rau 14 hnub sib law liag. Cov pab pawg ib txwm raug txhaj nrog tib koob tshuaj saline ib txwm. Cov tsos mob ntawm cov nas nyob rau hauv txhua pab pawg tau raug soj ntsuam los txiav txim seb lawv puas pom cov tsos mob xws li ntshai tsam khaub thuas, nyiam sov sov, txias txias, tso zis ntshiab thiab ntev, curled thiab arched rov qab, dull thiab ntxhov plaub hau, thiab kev nyuaj siab. Nyob rau tib lub sijhawm, qhov hnyav ntawm lub cev, tus naj npawb ntawm cov kev ua ub no, kev noj zaub mov, kev noj dej, tso zis tso zis thiab lwm yam ntsuas ntawm cov nas hauv txhua pab pawg tau ntsuas.

(3) Kev tswj hwm: Qhov ntau npaum li cas ntawm nas raug xam raws li kev kho mob ntau npaum ntawm Jinkui Shenqi Pills thiab Cistanche deserticola. Cov koob tshuaj ntawm txhua pawg Cistanche deserticola yog 1.80 g / kg raws li cov tshuaj nyoos. Qhov ntau npaum li cas yog qhia nyob rau hauv Table 2. Txhua extract ntawm Cistanche deserticola thiab Jinkui Shenqi Pills tau npaj rau hauv cov kev daws teeb meem ntawm ib co concentration nrog deionized dej. Cov tshuaj tau muab tshuaj intragastrically txhua hnub nrog ib qho intragastrically ntim ntawm 10 mL / kg rau 30 hnub sib law liag. Cov pab pawg ib txwm muaj thiab cov qauv pab pawg tau tswj hwm intragastrically nrog qhov sib npaug ntawm cov dej deionized txhua hnub. Lub cev hnyav tau ntsuas txhua lub lim tiam, thiab qhov ntim intragastrically tau hloov kho raws li lub cev hnyav ntawm cov nas. Tom qab kev tswj hwm, cov nas tau yoo mov rau 12 teev. Tom qab pentobarbital anesthesia, cov ntshav tau sau los ntawm lub plab aorta rau kev kuaj ntshav index. Lub raum, noob qes, epididymis, seminal vesicles thiab prostate raug tshem tawm kom pom qhov ntsuas ntsuas.