Efficiency Thiab Mechanisms Ntawm Angelica Sinensis Cistanche Fiber Compound Rau cem quav

Abstract:cem quavyog ib qho kab mob loj heev uas yuav cuam tshuam rau tib neeg lub neej zoo thiab ua rau muaj kev pheej hmoo kis kab mob. Nyob rau hauv txoj kev tshawb no, achronic constipation nas qauv yog tsim los siv loperamide hydrochloride.Mice tau gavaged ib Angelica sinensis Cistanche Fiber Compound, muaj Dang Gui [Angelica sinensis (Oliv.) Diels],Roob Cong Rong (Cistanche deserticolaY.C.Ma), nplej fiber ntau, thiab qis-molecular-yuag xylan ntawm qhov ntau npaum li cas (3.6 g·kg-1, 30 npaug ntawm qhov pom zoo rau tib neeg koob) thiab noj tsawg (0.6 g·kg{{8}) }, 5 zaug pom zoo tib neeg koob) rau 14 hnub. Txoj kev tshawb no tau soj ntsuam cov kev kho mob ntawm cov tshuaj los ntawm kev soj ntsuam fecal morphology, ntsuas dej cov ntsiab lus, thiab ua cov kev sim me me ntawm cov hnyuv. Tsis tas li ntawd, enzyme-linkedimmunosorbent assays (ELISA) tau ua los ntsuas cov qib ntshav ntawm cov tshuaj hormones hauv plab nrog rau motilin (MTL), gastrin (GAS), inflammatoryfactors interleukin -1 (IL-1 ), thiab qog necrosis factor. - (TNF- ). Kev kuaj mob histopathological nrog H&E staining tau ua haujlwm los ntsuas cov ntaub so ntswg puas tsuaj. Tsis tas li ntawd, Western blot thiab immunohistochemistry thwmsim tau ua los tshuaj xyuas cov qib qhia ntawm aquaporin-3 (AQP3) thiab c-Kit hauv txoj hnyuv. Theresults tau qhia tias Angelica sinensis Cistanche Fiber Compound tuaj yeem txhim kho fecal morphology, nce fecal cov ntsiab lus, thiab txhim kho cov hnyuv me me hauv cov nas. Tsis tas li ntawd, muaj kev nce qib tseem ceeb hauv cov ntshav qab zib cov tshuaj hormones thiab kev txo qis hauv cov qib ntawm inflammatoryfactors. Kev txhim kho hauv kev puas tsuaj ntawm cov kab mob hauv cov kab mob hauv colonic histopathological tau nrog los ntawm kev txo qis hauv kev qhia ntawm cov kab mob hauv cov dej hauv cov kab mob protein AQP3 thiab qhov tseem ceeb nce hauv c-Kit qhia. Cov txiaj ntsig no tau sib sau ua ke qhia tias muaj kev sib raug zoo ntawm koob tshuaj. Cov kev tshawb pom no qhia tau hais tias Angelicasinensis Cistanche Fiber Compound zoo txo ​​qis cem quav hauv nas. Nws cuam tshuam nrog kev tswj hwm ntawm cov kab mob hauv cov dej hauv cov kua dej AQP3 thiabc-Kit qhia, nrog rau kev hloov pauv ntawm cov ntshav qabzib thiab cov tshuaj hormones hauv plab. Qhov kev sim no tau pom zoo los ntawm Animal ExperimentEthics Committee ntawm Jinan University.

Ntsiab lus:cem quav; plab hnyuv lubrication thiab cem quav nyem; khoom noj khoom haus; txoj hnyuv; tshuaj suav tshuaj

Nws yuav siv sij hawm ntev npaum li cas rau CISTANCHE ua haujlwm?

cem quavyog akab mob gastrointestinal. The global prevalence of constipation in the elderly (>60 xyoo) yog 18.9% [1], thiab feem ntau ntau dua rau cov poj niam [2]. Nws yog tus cwj pwm los ntawm qhov nyuaj ntawm qhov defecation thiab nquag cem quav. txo thiab tawv tawv ntawm cov quav, thiab lwm yam, ua rau lub cev tsis xis nyob thiab plab hnyuv, thiab txawm inducing los yog aggravating gastrointestinal kab mob, kab mob plawv [3], thiab lwm yam., cuam tshuam rau lub neej zoo. Cov tshuaj suav tshuaj suav tshuaj tau dav siv los txhim kho cem quav thiab muaj txiaj ntsig zoo thiab paub tseeb kho mob [4,5]. Angelica thiab Cistanche deserticola fiber ntau siv hauv txoj kev tshawb no yog npaj los ntawm Cistanche deserticola, Angelica sinensis, nplej fiber ntau, thiab xylooligosaccharides. Cistanche deserticola thiab Angelica sinensis yog ob hom tshuaj suav tshuaj suav tshuaj hauv cov tshuaj laxatives, uas muaj cov txiaj ntsig ntawm kev tswj cov hnyuv ua haujlwm thiab txhim kho cem quav. [6-9], ob hom tshuaj suav tshuaj muaj keeb kwm tib yam li tshuaj thiab zaub mov hauv kuv lub tebchaws. Lawv siv ua ke tuaj yeem ua tiav cov txiaj ntsig ntawm kev ntxiv qi thiab ntshav thiab kev sib haum xeeb yin thiab yang. Nplej fiber ntau yog nplua nuj nyob rau hauv insoluble thiab soluble noj fiber ntau, uas yuav txhawb txoj hnyuv peristalsis thiab defecation, thiab feem ntau yog siv los ua ib qho tshuaj pab los kho cem quav; xylo-oligosaccharide yog ib qho kev ua haujlwm ntawm oligosaccharide uas tuaj yeem tswj cov kab mob hauv plab, yog li txhawb txoj hnyuv peristalsis. , nce moistness ntawm cov quav [10], muab cov plaub qhov tsw, thiab kho cov hauv paus hniav ntawm tus kab mob. Txoj kev tshawb no tsom los ntsuas qhov ua tau zoo ntawm cov angelica thiab cistanche deserticola fiber ntau ntawm cov nas cem quav thiab tshawb nrhiav nws txoj kev ua tau zoo.

Cov ntaub ntawv thiab cov txheej txheem

Kev sim tsiaj: 40 yam kab mob tsis muaj kab mob (SPF) qib txiv neej Kunming nas, hnub nyoog 4 txog 5 lub lis piam, tau yuav los ntawm Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., daim ntawv tso cai naj npawb SCXK (Guangdong) 2021-0011, The ambient kub yog 22 ± 2 degree, cov av noo yog 50% ± 10%, thiab nruab hnub thiab hmo ntuj voj voog yog 12 h / 12 h. Cov txheej txheem kev sim thiab cov txheej txheem ua raws li kev coj noj coj ua ntawm kev siv tsiaj thiab kev saib xyuas thiab tau txais kev pom zoo los ntawm Pawg Saib Xyuas Tsiaj Txhaum Cai ntawm Jinan University.

Cov tshuaj ntsuam xyuas: Angelica sinensis thiab Cistanche deserticola fiber compound npaj yog tsim los ntawm Angelica Sinensis thiab Cistanche deserticola, uas yog muab rho tawm nrog dej, concentrated, qhuav, thiab crushed kom tau ib tug qhuav paste hmoov (lub paste yield yog 52.95%). Cov hmoov nplej qhuav yog tov nrog cov nplej fiber ntau thiab xylo-oligosaccharide rau kev siv tom qab. Angelica sinensis (batch tooj 230404CP108) thiab Cistanche deserticola (batch tooj 230303CP109) tau yuav los ntawm Hebei Wanxiu Pharmaceutical Co., Ltd., thiab tau sim ua raws li 2020 tsab ntawm Suav Pharmacopoeia. Xylo-oligosaccharides tau yuav los ntawm Shandong Bailong Chuangyuan Biotechnology Co., Ltd. Tus lej batch yog 20210829026, thiab kev tshuaj xyuas ua raws li GB / T35545; cov nplej fiber ntau los ntawm J. Rettenmaier & Söhne GmbH + Co KG, thiab tus lej batch yog 07501230126. Raws li tus neeg laus 60 kg, qhov pom zoo noj ntawm angelica thiab cistanche fiber ntau rau tib neeg yog 7.118 g·d-1 , ntawm qhov ntau npaum li cas ntawm cov nplej fiber ntau yog 3.5g·d-1. Lub compatibility ntawm no compound yog angelica: cistanche: nplej fiber ntau: xylooligosaccharides=2: 2: 3: 2 (qhov ntau npaum li cas ntawm decoction pieces yog raws li cov tshuaj crude). Noj 30 npaug ntawm cov neeg tau txais kev pom zoo raws li pawg koob tshuaj siab thiab 5 npaug ntawm cov tib neeg pom zoo noj raws li pawg neeg tsawg, raws li 3.6 thiab 0.6 g·kg-1·d-1. Npaj ib qho kev ncua nrog dej ntshiab, sib tov zoo, thiab tso tseg.

Tshuaj thiab Reagents Loperamide Hydrochloride (loperamide, LOP), specifications: Txhua ntsiav tshuaj muaj loperamide hydrochloride

Amine 2 mg, 12 capsules/box, khoom batch naj npawb: MIJ3102, muas los ntawm Xi'an Janssen Pharmaceutical Co., Ltd.; Mosapride Citrate Tablets (Quali), specifications: Txhua ntsiav tshuaj muaj 5 mg ntawm Mosapride Citrate, 24 ntsiav tshuaj / lub thawv, khoom batch naj npawb: 25220603, muas los ntawm Lunan Beite Pharmaceutical Co., Ltd.; activated carbon hmoov, specification: 200 mesh xim av hmoov, batch naj npawb A805337, muas los ntawm Shanghai McLean Biochemical Technology Co., Ltd.; Cov pos hniav arabic, specification: hmoov, batch naj npawb A108975, yuav los ntawm Shanghai Aladdin Biochemical Technology Co., Ltd.

Tsis muaj ethanol (Tianjin Damao Chemical Reagent Factory, 64-17-5); Ammonium persulfate (Shanghai McLean Biochemical Technology Co., Ltd., A6295-500g); Tris-Base (FD2010), 5 × loading buffer (FD002), -actin antibody (FD0060), tshis tiv thaiv luav theem nrab antibody (FDR007), tshis tiv thaiv nas thib ob antibody (FDM007), thiab ECL xim kev loj hlob cov khoom (FD8020) tau tag nrho yuav los ntawm Hangzhou Fude Biotechnology Co., Ltd.; RIPA lysis buffer (Shanghai Biyuntian Biotechnology Co., Ltd., P0013B); 4% paraformaldehyde (Biosharp tuam txhab, BL539A); Cov khoom siv Immunohistochemistry (Jiangsu KGI Biotechnology Co., Ltd., KGOS60); bicinchoninic acid (BCA) Protein quantification kit (Thermo Fisher Scientific, SLZ60212); motilin (MTL), gastrin (GAS), thiab inflammatory factor interleukin -1 (IL-1), qog necrosis factor (tumor necrosis factor- , TNF- ) ELISA cov khoom (Shanghai Enzyme Biotechnology Co., Ltd. ., YJ201829, YJ037432, YJ002095, YJ301814); aquaporin 3 (aquaporin 3, AQP3) antibody (Affinity Company, AF5222); c -Kit antibody (Cell Signaling Technology, 3074S).

Khoom siv: Super ntshiab dej ntsuas (Millipore Company, USA, Milli Q); Electronic Balance (Sartorius, Lub teb chaws Yelemees)

Lub tuam txhab, BS224S); Electronic analytical tshuav nyiaj li cas (Shanghai Benpu Instrument Technology Co., Ltd., FA2204N); High-speed refrigerated centrifuge (Sigma tuam txhab, lub teb chaws Yelemees, 3K15); Microplate nyeem ntawv (Thermo Company, USA, Multiskan Mk 3); Paraffin microtome (Leica, Lub Tebchaws Yelemees) Lub Tuam Txhab, RM2255); Kev kub ceev hlau da dej (Shanghai Yiheng Technology Co., Ltd., TU-100C); Tsis siv neeg qauv ceev ceev grinder (Shanghai Jingxin Technology Co., Ltd., JXFSTPRP-24); Electrophoresis ntsuas (Beijing Liuyi Biotechnology Co., Ltd., DYY-6C); scanning microscope (Precipoint Company, Precipoint M8); chemiluminescence imaging system (Shanghai Tianneng Technology Co., Ltd., Tanon 5200).

Kev koom ua pawg thiab kev tsim qauv: 40 nas tau hloov kho rau 5 hnub thiab muab faib ua pawg tswj tsis zoo.

(tswj), pawg qauv (LOP), pawg siab thiab qis koob tshuaj (LOP + PF-H, OP + PF-L), pawg tswj zoo (mosapride citrate, LOP + MOS). Thaum 9 txog 10 sawv ntxov txhua hnub, txhua pab pawg tshwj tsis yog pawg tswj hwm tsis zoo tau muab 10 mg·kg-1 loperamide hydrochloride los ntawm gavage los tsim tus qauv cem quav, thiab pab pawg tswj tsis zoo tau muab rau sib npaug ntawm cov dej ntshiab los ntawm gavage. Los ntawm 14 mus rau 15 teev, cov pab pawg muaj koob tshuaj ntau thiab cov koob tshuaj qis tau muab cov tshuaj sib xyaw 3.6 thiab 0.6 g·kg-1 raws li, thiab pawg tswj hwm zoo tau muab mosapride citrate (MOS) 5 mg·kg{{ 19}} ib.

, pawg tswj tsis zoo thiab pawg tswj hwm tus qauv tau muab sib npaug ntawm cov dej ntshiab los ntawm gavage rau 14 hnub sib law liag.

Cov xwm txheej thiab cov quav ntawm cov nas tau pom thiab sau tseg thaum lub sijhawm siv tshuaj. Cov xwm txheej thiab cov quav ntawm nas nyob hauv txhua pab pawg tau sau tseg.

Cov cim, hnyav 3 zaug hauv ib lub lis piam thiab kho qhov ntau npaum li cas. Hnub 3, 7, 10, thiab 14 tom qab kev tswj hwm, 3 mus rau 4 pellets ntawm cov quav tshiab los ntawm txhua tus nas tau sau, ntsuas tam sim ntawd, thiab sau cia li ntub qhov hnyav, qhuav hauv qhov cub, thiab sau cia li qhuav qhov hnyav, thiab fecal noo noo. cov ntsiab lus raug xam. Xam cov fecal noo noo cov ntsiab lus (%)=(ntub qhov hnyav − qhov hnyav qhuav) / ntub dej × 100%.

Kev txiav txim siab ntawm txoj hnyuv me me ua haujlwm tau ua raws li cov kev sim uas yuav tsum tau ua los ntawm cov ntaub ntawv siv [11] thiab "Txoj Kev Ntsuas thiab Kev Ntsuas Kev Ua Haujlwm ntawm Cov Khoom Noj Noj Qab Haus Huv (2023 tsab)". Tom qab koob tshuaj zaum kawg, cov nas hauv txhua pab tau yoo mov thiab dej tsis pub dhau 16 teev. Cov qauv pawg, cov qauv sib xyaw Cov pab pawg siab- thiab qis qis thiab pab pawg tswj tau zoo tau muab loperamide hydrochloride ncua kev kawm (10 mg·kg-1) los ntawm gavage, thiab pawg tswj tsis zoo tau muab rau sib npaug ntawm cov dej ntshiab los ntawm gavage. 0.5 teev tom qab ntawd, cov tshuaj sib xyaw ua ke ntawm cov koob tshuaj siab thiab cov tshuaj qis qis thiab cov pab pawg tswj tau zoo tau muab tus cwj mem (muaj 5%) uas muaj cov koob tshuaj coj mus kuaj (3.6 g·kg-1, 0.6 g·kg-1, 5 mg·kg-1) raws. (activated charcoal hmoov, 10% cov pos hniav arabic), thiab sib npaug ntawm tus number case tau muab rau cov pawg tsis zoo thiab cov qauv. Cov tsiaj raug tua tam sim ntawd tom qab 0.5 h los ntawm lub ncauj tsev menyuam dislocation, thiab "tag nrho qhov ntev ntawm cov hnyuv" thiab "ink advancement length" raug ntsuas. Xam tus number case nce qib=ink advancement length (cm)/tag nrho qhov ntev ntawm cov hnyuv (cm) × 100%.

Txhawm rau txheeb xyuas cov qib ntawm MTL, GAS, TNF- , thiab IL-1 hauv cov ntshav, cov ntshav tau muab los ntawm cov nas pob qhov muag, centrifuged ntawm 4 degree, 3 500 r·min-1 rau 10 feeb, cov ntshav siab tau muab coj los ua, thiab MTL thiab GAS hauv cov ntshav nas tau txiav txim siab raws li cov khoom siv ELISA. thiab TNF- , IL-1 qib.

H&E kuaj pom cov kev hloov pauv ntawm cov kab mob hauv cov hnyuv. Cov nyuv ntawm nas yog ntxuav nrog ib txwm saline thiab incubated nyob rau hauv 4% paraformaldehyde.

Lawv tau kho nyob rau hauv aldehyde, embedded ib txwm tom qab 24 teev, muab faib, ci, thiab H&E stained, thiab cov pathological hloov ntawm cov nyuv cov ntaub so ntswg raug soj ntsuam nyob rau hauv lub teeb lub teeb.

Cov txheej txheem immunohistochemical tau siv los txheeb xyuas qhov kev qhia ntawm AQP3 thiab c-Kit hauv txoj hnyuv. Cov ntaub so ntswg ntawm cov nas los ntawm txhua pawg tau sau.

Paraffin seem, dewaxing, hydration, antigen retrieval, blocking endogenous catalase, thaiv nrog 2% tshis serum, ntxiv AQP3 thiab c-Kit antibodies feem, thiab incubating hmo ntuj ntawm 4 degree, phosphate buffered saline, PBS) rau 5 min Hematoxylin countertains ethanol lub cev qhuav dej, xylene pob tshab, nruab nrab cov pos hniav sealing, scanning nyob rau hauv lub tshuab kuaj kab mob, thiab siv Image J software los soj ntsuam cov txiaj ntsig.

Western blot txoj kev tau siv los txheeb xyuas qhov kev qhia ntawm AQP3 thiab c-Kit hauv txoj hnyuv. Noj 10 mg ntawm cov nqaij khov nab kuab thiab muab tso rau hauv ib lub raj centrifuge nrog 150 μL ntawm RIPA lysis tsis muaj 1% protease inhibitor. Siv lub tshuab grinder ceev kom homogenize cov ntaub so ntswg. , centrifuge lub homogenate nrog ib tug tsawg-kub centrifuge ntawm 10 000 r·min-1 rau 10 feeb, coj lub supernatant, txiav txim siab cov protein concentration los ntawm BCA protein quantification txoj kev, sib tov nrog lub loading tsis, denature cov protein, thiab txheeb xyuas cov txiaj ntsig raws li txhua tus qauv. 30 ug protein tau thauj khoom, raug rau SDS-PAGE electrophoresis, ntub dej hloov mus rau daim nyias nyias, thiab tom qab ntawd thaiv nrog 5% skimmed mis nyuj hmoov nyob rau hauv chav tsev kub rau 1.5 h, ntxuav peb zaug nrog TBST, tso thawj cov tshuaj tiv thaiv ntawm lub shaker thiab incubated thaum hmo ntuj. 4 degree, ces tshem tawm, thiab ntxuav nrog TBST tsis ntxuav 3 zaug nrog tov, incubate nrog lwm yam tshuaj tiv thaiv rau 1 h ntawm chav tsev kub thiab ces ntxuav 3 zaug nrog TBST. ECL chemiluminescence ua hauj lwm tov yog siv rau kev loj hlob xim, thiab ib tug tag nrho tsis siv neeg chemiluminescence imaging system yog siv rau txoj kev loj hlob. Image J software yog siv los txheeb xyuas cov txiaj ntsig.

Cov txheej txheem txheeb cais Tag nrho cov ntaub ntawv koom nrog hauv qhov kev sim no tau nthuav tawm hauv daim ntawv txhais tau tias ± tus qauv sib txawv. GraphPad Prism tau siv los npaj cov ntaub ntawv sim, thiab kev sib piv ntawm cov pab pawg tau ua tiav siv ib txoj kev ANOVA lossis ob txoj kev ANOVA; Thaum qhov sib txawv ntawm cov pab pawg tau txheeb xyuas qhov tseem ceeb, qhov sib txawv tsawg kawg nkaus (LSD) txoj kev siv. ) Ua kev sib piv ua ke ntawm cov pab pawg, thiab P < 0.05 yog suav tias yog qhov sib txawv tseem ceeb.