PART 1 Anti-Osteoporotic Activity Of An Edible Traditional Chinese Medicine Cistanche Deserticola Ntawm Cov Pob Txha Metabolism Ntawm Ovariectomized nas Los ntawm RANKL/RANK/TRAF6- Mediated Signaling Pathways

Bo Zhang 1†, Ling-Ling Yang 1 †, Shu-Qin Ding 1 †, Jing-Jing Liu 1, Yan-Hong Dong 1,

Yan-Ting Li 1, Nan Li 1, Xiao-Jun Zhao 1, Chang-Ling Hu 2, Yiping Jiang 3* and Xue-Qin Ma 1*

1 Lub Chaw Haujlwm Tseem Ceeb ntawm Hui Ethnic Medicine Modernization, Department of Pharmaceutical Analysis, Ministry of Education, Lub Tsev Kawm Ntawv ntawm Pharmacy, Ningxia Medical University, Yinchuan, Tuam Tshoj, 2 Lub Chaw Haujlwm Saib Xyuas Khoom Noj thiab Tib Neeg Kev Noj Qab Haus Huv, Center for Excellence in Post-Sau Technologies, North Carolina Tshawb fawb Campus, North Caroline A&T State University, Greensboro, NC, United States, 3 Department of Pharmacognosy, The Second Military Medical University, Shanghai, Suav

cistanchekho tauosteoporosis

Muab cov kev txwv ntawm cov kws kho mob uas twb muaj lawm rau kev kho mob osteoporosis postmenopausal osteoporosis, tseem tseem xav tau kev xaiv ntau ntxiv nrog rau ob qho tib si ua tau zoo thiab muaj kev cuam tshuam tsawg dua.Cistanche deserticolaYC Ma yog lub npe hu ua cov tshuaj tonic nrov ib txwm siv los kho lub raum tsis muaj zog nrog rau cov leeg tsis muaj zog hauv cov neeg tsawg hauv cov nroog Esxias. Raws li txoj kev xav ntawm "lub raum dominate pob txha," ib qho ovariectomized (OVX) nas qauv ntawm postmenopausal osteoporosis tau siv los ntsuas cov kev kho mob ntawmC. deserticolaextract (CDE) ntawm pob txha poob. Plaub caug-yim tus poj niam Sprague-Dawley nas, hnub nyoog li ntawm 12 lub lis piam, tau random muab tso rau hauv rau pawg, suav nrog cov sham pab pawg neeg hais lus nrog 0.5 feem pua ​​carboxymethyl cellulose sodium (CMC-Na) (sham), pawg zoo kho nrog 1 mg / kg ntawm estradiol valerate (EV), qis, nruab nrab, thiab siab ntau pab pawg neeg hais lus nrog 200, 400, thiab 800 mg / kg / hnub ntawm CDE, feem. Tom qab 3 lub hlis ntawm kev cuam tshuam tas mus li, CDE tau nthuav tawm cov haujlwm tseem ceeb los tiv thaiv osteoporotic ua pov thawj los ntawm kev txhim kho tag nrho cov pob txha pob txha pob txha, ameliorated pob txha microarchitecture; nce alkaline phosphatase kev ua haujlwm; txo deoxypyridinoline, cathepsin K, tartrate-resistant acid phosphatase, thiab malondialdehyde ntau ntau; thaum lub cev, lub tsev menyuam, thiab qhov chaw mos qhov hnyav hauv OVX nas tsis cuam tshuam los ntawm CDE kev cuam tshuam. Tsis tas li ntawd, qhov tshwm sim zoo li tsis sib haum xeeb ntawm qib calcium thiab phosphorus ntawm OVX thiab sham nas tau pom thiab piav qhia. Mechanistically, CDE txo qis-tswj cov qib ntawm TRAF6, RANKL, RANK, NF-κB, IKK, NFAT2, thiab nce- tswj cov phosphatidylinositol 3-kinase (PI3K), AKT, osteoprotegerin, thiab c-Fos kab lus, uas txhais tau hais tias CDE tuaj yeem cuam tshuam RANKL / RANK-induced activation ntawm downstream NF-κB thiab PI3K / AKT txoj hauv kev, thiab thaum kawg, tiv thaiv kev ua ntawm cov tseem ceeb osteoclastogenic proteins NFAT2 thiab c-Fos. Tag nrho cov ntaub ntawv qhia tias CDE muaj peev xwm tiv thaiv osteoporotic kev ua ub no thiab cov nyhuv no yog, tsawg kawg yog ib feem, koom nrog kev hloov kho ntawm RANKL/RANK/TRAF6-kho NF-κB thiab PI3K / AKT signaling nrog rau c- Fos thiab NFAT2 qib. Yog li, CDE tuaj yeem sawv cev rau tus neeg sib tw muaj txiaj ntsig zoo rau kev kho mob osteoporosis postmenopausal.

Ntsiab lus:Cistanche deserticola, antiosteoporotic, TRAF6, RANKL, RANK

Yog xav paub ntxiv thov hu rau:Joanna.jia@wecistanche.com

Cistanche deserticola muaj ntau yam teebmeem, nyem qhov no kom paub ntau ntxiv

Taw qhia

Txij li thaum osteoporosis tsis tau suav tias yog ib feem ntawm kev laus ib txwm muaj, nws tau txais kev nce qib hauv cov pob txha pob txha. Niaj hnub no, osteoporosis tau dhau los ua kev phom sij loj rau kev noj qab haus huv uas cuam tshuam ntau dua 200 lab tus tib neeg thoob plaws ntiaj teb nrog cov neeg tuaj koom 10 billion nyiaj tau siv txhua xyoo rau cov tib neeg no (Ma et al., 2015). Osteoporosis, suav nrog postmenopausal thiab senile osteoporosis, yog tus cwj pwm los ntawm micro-architectural deterioration thiab cov pob txha pob txha tsawg (BMD) vim qhov tsis sib xws ntawm osteoblast-mediated pob txha tsim thiab osteoclast-mediated pob txha resorption, ntsig txog (Kanis li al. ). Nyob rau hauv kev txiav txim siab ntawm cov kev txwv ntawm cov kev xaiv kho tam sim no thiab qee qhov kev tsis zoo ntawm cov khoom siv hluavtaws (Ye et al., 2015) rau kev kho cov kab mob osteoporosis, nws yog ib qho tsim nyog rau lwm txoj hauv kev uas muaj txiaj ntsig zoo thiab muaj kev phiv tsawg (Tan li al., 2017). Raws li ib qho zoo kawg thiab lwm txoj kev kho mob rau kev kho mob ntawm ntau yam kab mob xws li postmenopausal osteoporosis, cov tshuaj ntsuab tshwj xeeb tshaj yog cov tshuaj suav tshuaj (TCM) tau maj mam lees paub thiab tau lees paub (Czerwinska thiab Melzig, 2018; Ma et al., 2018). Ob txhiab xyoo dhau los,Cistanche deserticolaYC Ma tau pom muaj txiaj ntsig zoo rau tib neeg kev noj qab haus huv thiab tau sau tseg hauv phau ntawv suav tshuaj suav tshuaj npe hu ua Shen Nong Herbal (Anonymous, 1981). Tam sim no, nws yog ib qho tseem ceeb TCM tau sau tseg nyob rau hauv Suav pharmacopeia (Suav thiab Pharmacopoeia, 2015) thiab kuj yog ib tug nto moo tonic tshuaj ntsuab uas tau siv nyob rau hauv Asian counties nrog rau Tuam Tshoj thiab Nyiv rau ntau pua xyoo (Wong et al., 2006) .C. deserticolatau pom tias muaj kev nyab xeeb zoo (Liao li al., 2018) thiab dav siv tshuaj kho mob: hauv pej xeem, nws tau dav siv los ua noj nrog ntau hom tshuaj ntsuab cuisine rau kev kho mob raum tsis txaus, cov leeg tsis muaj zog, thiab lumbar tsis muaj zog. ; Niaj hnub no, kev saib xyuas ntau ntxiv tau them rau nws ntau yam tshuaj siv tshuaj xws li tshuaj tiv thaiv kab mob, tiv thaiv qaug zog, tshuaj tiv thaiv kab mob, tshuaj tiv thaiv kab mob antioxidant, txhim kho kev tiv thaiv kab mob, thiab lwm yam (Gu et al., 2016; Fu li al., 2018). Nws tseem yog lub npe hu ua "ginseng ntawm cov suab puam" hauv Suav teb vim nws cov tshuaj kho mob zoo heev. Ob qho tib si thiab niaj hnub daim ntawv thov ntawmC. deserticolaua cov tshuaj ntsuab no nrov nyob rau hauv ob qho tib si tshuaj thiab kev noj qab haus huv kev lag luam, yog li tau raug tsim los rau hauv cov tshuaj thiab zaub mov kua pom zoo los ntawm lub Xeev Food and Drug Administration. MuabC. deserticolafeem ntau yog ua hauj lwm los daws lub raum tsis muaj peev xwm hauv Suav pej xeem cov tshuaj, uas txhais tau hais tias cov tshuaj ntsuab no tuaj yeem suav tau tias yog ib tus neeg sawv cev zoo tshaj plaws los cuam tshuam cov kab mob osteoporosis raws li kev xav ntawm "lub raum dominate pob txha" (Wang li al., 2016). Cov ntaub ntawv luam tawm tau ua pov thawj cov nyhuv anti-osteoporotic ntawm C. deserticola extracts ob leeg hauv vivo thiab hauv vitro (Liang li al., 2011; Li et al., 2012; Liang et al., 2013; Xu et al., 2017; Nkauj li al., 2018), thiab ntau lub tebchaw cais xws li echinacoside (Li et al., 2013), acteoside (Lee et al., 2013), thiab cistanoside A (Xu et al., 2017) uas kuj tau tshaj tawm txog kev ua haujlwm tiv thaiv. - osteoporotic kev ua ub no; thiab lwm cov tebchaw xws li 2'-acetylacteoside tau lees paub tias muaj cov tshuaj tiv thaiv kab mob antioxidant, tiv thaiv kab mob, neuroprotective, hepatoprotective, kev tiv thaiv kab mob (Li li al., 2016), thiab kev tiv thaiv kev laus (Peng li al., 2016). Txog rau hnub tim, kaum ob ntawm bioactive phenylethanoid glycosides tau raug txheeb xyuas los ntawmCistanche tshuaj ntsuab(Wang li al., 2012), echinacoside, thiab acteoside yog cov ntsiab lus tseem ceeb uas muaj nyob hauv feem ntau ntawm Cistanche hom nrog cov ntsiab lus yog 1.83–41.49 thiab 0.27–8.28 mg/kg , raws li; whereas lwm cov phenylethanoid glycosides suav nrog 2'-acetylacteoside, 6'-acetylacteoside, cistanoside A, cistanoside C, thiab isoacteoside yog 1.56–3.16 mg/g, 0.49–1.66 mg/kg, 1.41–1{ 29}}.11 mg/kg, 0.33–2.24 mg/kg, thiab 0.08–5.00 mg/kg, ntsig txog (Dong et al., 2018). Raws li cov ntaub ntawv tshaj tawm saum toj no, peb pom tias cov hauv qab los tiv thaiv osteoporotic molecular mechanisms ntawmC. deserticolatshwj xeeb tshaj yog qhov kev taw qhia txog dej ntws tsis tau meej; thiab cov compound nws tus kheej tsis tuaj yeem sawv cev rau cov txiaj ntsig ntawm cov tshuaj ntsuab uas muaj cov cuab yeej sib koom ua ke los ntawm ntau hom khoom sib txawv; Tsis tas li ntawd, cov hom phiaj tshwj xeeb ntawm kev taw qhia txoj hauv kev tau sib txawv ntawm cov khoom sib txawv. Txoj haujlwm tam sim no tsom rau kev tshawb xyuas qhov muaj peev xwm tiv thaiv tau ntawmC. deserticolatawm tsam ovariectomized (OVX) nas nyob rau hauv vivo nrog rau qhov tseem ceeb ntawm txoj cai ntawm qog necrosis yam receptor-associated factor 6 (TRAF6), receptor activator ntawm nuclear factor kappa B ligand (RANKL), RANK qhia thiab RANKL/RANK/TRAF{{3} } induced nuclear factor kappa B (NF-κB), thiab phosphoinositide 3-kinase (PI3K)/protein kinase (AKT) signalings. Kev kho pob txha, suav nrog kev tsim kho thiab kho, yog ib qho txheej txheem sib txawv uas tswj hwm los ntawm qhov sib npaug ntawm cov pob txha osteoblastic thiab osteoclastic pob txha resorption, thiab nce pob txha resorption feem ntau tshwm sim los ntawm kev txhim kho osteoclastogenesis feem ntau ua rau osteoporosis txawm pob txha puas. Feem ntau ntawm cov xov tooj ntawm tes, cov tshuaj hormones, thiab kev loj hlob yam tseem ceeb rau kev sib txawv ntawm osteoclast thiab kev ua haujlwm tau tswj hwm los ntawm RANK thiab nws ob lub ligands, RANKL thiab osteoprotegerin (OPG) (Lacey li al., 1998). Thiab qhov kev tshawb pom tsis ntev los no tau tshawb pom ntxiv TRAF6 yog qhov tseem ceeb ntawm kev tswj hwm hauv RANKL / RANK ua rau muaj teeb meem tshwm sim (Tan li al., 2017). Los ntawm kev nrhiav neeg ua haujlwm ntawm TRAF6, RANKL khi rau nws cov receptor RANK, thiab tom qab ntawd cov kev taw qhia qis qis suav nrog NF-κB thiab PI3K / AKT tau tshwm sim, tom qab ntawd qhov tseem ceeb osteoclastogenic cov proteins ntawm c-Fos thiab cov khoom siv nuclear ntawm activated T hlwb c2 (NFAT2) ( Takayanagi, 2007) tau tswj hwm, thiab thaum kawg, qhov sib txawv ntawm osteoclast tau pib. Nyob rau hauv txoj kev tshawb no, lub hauv paus ntawm kev tiv thaiv osteoporotic mechanism ntawm CDE ntawm RANKL/RANK/TRAF6-kho cov pob txha resorption yog soj ntsuam thiab sib tham.

Cov khoom siv thiab cov txheej txheem

Tsiaj txhu, Cells, Antibodies, thiab Reagents

Poj niam laus Sprague-Dawley nas (Ningxia Medical University, Yinchuan, Suav); RAW264.7 hlwb (Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, Tuam Tshoj); macrophage colony stimulating factor (M-CSF) thiab RANKL (PerroTech, Inc. USA); estradiol valerate [estradiol valerate (EV), Delpharm Lille SAS, Paris, Fabkis]; cathepsin K (CK) ELISA cov khoom siv (BioVision, American); deoxypyridinoline (DPD) thiab pob txha gla-protein (BGP) cross-links ELISA cov khoom siv (Xinyu Biological Engineering Co., Ltd, Shanghai, Suav); malondialdehyde (MDA), glutathione (GSH), thiab superoxide dismutase (SOD) reagent kits (Jianchen Biological Engineering, Nanjing, Suav teb); penicillin thiab streptomycin (Gibco, USA); polyvinylidene fluoride (PVDF) membrane (Millipore Life Sciences, USA); thawj cov tshuaj tiv thaiv suav nrog GAPDH (18AF0401), -actin (17AV0411), IκB kinase (IKK) (AD01134589), TRAF6 (2), RANKL (GR3193842-5), RANK (AA02113656), NFκBIA (32K261), AC09021266), OPG (AG06292776), c-Fos (AG12059411), AKT (AF05173234), NFAT2 (AO11015648), thiab lwm yam tshuaj tiv thaiv kab mob (horseradish peroxidase-conjugated tshis los tiv thaiv-Lavbit5, immunoglobulin) tau muab , Suav teb); Dulbecco's modified eagle's medium (DMEM), bicinchoninic acid protein quantization assay kit, fetal bovine serum (FBS), thiab Dulbecco's phosphate buffered saline (PBS) tau yuav los ntawm Hylcone (Logan, UT, USA); echinacoside [kev ua haujlwm siab ua kua chromatography (HPLC) Ntau dua lossis sib npaug li 98 feem pua ​​, 11167-200503, National Institutes for Food and Drug Control, China]; acteoside (HPLC Ntau dua lossis sib npaug li 98 feem pua, AB0497, ALFA, Tuam Tshoj); isoacteoside (HPLC Ntau dua lossis sib npaug li 98 feem pua, AB17021202, ALFA, Tuam Tshoj); 2'-acetyl acteoside (HPLC Ntau dua lossis sib npaug li 98 feem pua, PRF7081243, Chengdu Pu Rui Fa Technology, Co., Ltd. China); cistanoside A, cistanoside C, thiab 6'-acetyl acteoside (HPLC Ntau dua lossis sib npaug li 95 feem pua, cais thiab ntxuav hauv peb lub chaw kuaj) (Xu li al., 2017).

Cov khoom siv cog thiab kev npaj

Qhuav stems ntawmC. deserticolaYC Ma tau yuav los ntawm Lub Nroog Yongning, Ningxia xeev thaum lub Cuaj Hlis 2015, qhov chaw tshwj xeeb ntawm kev xaiv cov nroj tsuag yog (106.026597, 38.262816). Cov tshuaj ntsuab tau txheeb xyuas los ntawm Prof. Ling Dong (chaw haujlwm ntawm kws tshuaj, Ningxia Medical University), nrog rau daim ntawv pov thawj (#20150901) muaj nyob rau hauv herbarium ntawm kev tshuaj ntsuam xyuas tshuaj. Tag nrho ntawm 10.0 kg qhuav thiab hmoovC. deserticolatau muab rho tawm los ntawm kev siv cov txheej txheem kub reflux, cov txheej txheem ntxaws raws li hauv qab no: cov kuab tshuaj yog 75 feem pua ​​​​ethanol, qhov piv ntawmC. deserticolaCov khoom siv rau cov kuab tshuaj yog 1: 8, lub sij hawm reflux tau kav ntev li 2 teev, thiab cov ntaub ntawv tau reflux rau peb zaug; Tom qab ntawd tag nrho cov filtrates tau muab sib xyaw ua ke thiab ua kom pom tseeb hauv qhov txo qis kom them taus 6.5 kgC. deserticolaethanol extracts (CDE). Rau cov tsiaj sim, {{0}}.5 feem pua ​​​​carboxymethyl cellulose sodium (CMC-Na) tau siv los tshem tawm CDE, thiab cov tshuaj noj hauv qhov ncauj tau teem tseg li 200, 400, thiab 800 mg / kg / hnub, nrog ntim ntawm 1 ml / 100 g lub cev hnyav ntawm txhua tus tsiaj; rau western blot tsom xam, dimethyl sulfoxide tau ua hauj lwm los yaj CDE thiab ces diluted nrog DMEM kom tau qhov kawg concentration ntawm 0.01, 0.1 1 mg/ml; rau HPLC tsom xam, methanol tau siv rau yaj CDE, ua raws li lim thiab diluted nrog thawj theem mobile kom tau raws li qhov xav tau.

High-Performance Liquid Chromatography Analysis ntawmCistanche deserticola Extract

Cov tshuaj lom neeg tseem ceeb ntawm CDE tau txheeb xyuas los ntawm kev siv HPLC nrog cov khoom siv sib xws. Cov xwm txheej chromatography tau teeb tsa raws li hauv qab no: Agilent 1220 RP-HPLC ntsuas nrog C18 kem (4.6 id × 250 mm; 5 μm), gradient elution mobile theem feem ntau muaj dej thiab acetonitrile siv. kom tau txais cov txiaj ntsig zoo sib cais: hnyav A (dej muaj 0.5 feem pua ​​​​acetic acid) thiab B (acetonitrile): 0–4 min, 85–83 feem pua ​​A; 4–10 feeb, 83–80 feem pua ​​A; 10–30 feeb, 80–75 feem pua ​​A; 30–40 min, 75–70 feem pua ​​A. Cov qauv txhaj tshuaj yog 5 ul, qhov kub thiab txias tau tswj ntawm 30 degree, thiab UV kuaj tau ua tiav ntawm 333 nm (Dong et al., 2018). Cov tshuaj siv xws li echinacoside, acteoside, isoacteoside, cistanoside A, cistanoside C, 2'-acetyl acteoside, thiab 6'-acetyl acteoside tau siv los txheeb xyuas qhov sib thooj.

cistancheua tauanti-apoptosis

Kev sim tsiaj

Rau cov tsiaj sim, tag nrho cov txheej txheem tau ua raws li cov txheej txheem pom zoo los ntawm lub koom haum saib xyuas tsiaj thiab siv pawg thawj coj ntawm Ningxia Medical University (NXMU{{0}}}). Plaub caug-yim tus poj niam laus Sprague-Dawley nas hnyav txog 237 ± 25 g tau txais (Ningxia Medical University, Yinchuan, Suav teb) thiab khaws cia rau 1 lub lis piam nrog kev noj zaub mov pellet thiab dej kais dej hauv ib puag ncig kev tswj hwm. Tom qab ntawd, txhua tus nas tau txhaj tshuaj loog (chloral hydrate, 100 mg / kg, ip) nkaus xwb, sham ovariectomized (sham), lossis ob lub zes qe menyuam raug tshem tawm thiab tom qab ntawd muab faib ua tsib pawg: pawg qauv (OVX) tau hais lus nrog 0.5. feem pua ​​CMC-Na; pawg zoo (EV) nrog 1 mg / kg / hnub ntawm estradiol valerate; qis (CDEL), nruab nrab (CDEM), thiab siab (CDEH) pawg noj tshuaj nrog 200, 400, thiab 800 mg / kg / hnub ntawm CDE, feem. Cov kev sim tau siv sijhawm li 12 lub lis piam; lub cev hnyav ntawm txhua tus nas raug ntsuas ob lub lis piam nrog kev tswj hwm koob tshuaj hloov raws li. Tom qab hnub kawg ntawm kev cuam tshuam, 24 teev cov zis, ntshav, femora, tibia, thiab lub tsev menyuam tau raug sau thiab khaws cia ntawm −80 degree rau kev txheeb xyuas ntxiv.

Biochemical Parameters

Cov qib ntawm serum SOD, GSH, MDA, cathepsin K, BGP, thiab urinary DPD tau txiav txim siab los ntawm cov khoom siv reagent, thaum cov kev ua ntawm tartrate-resistant acid phosphatase (TRAP) tau soj ntsuam raws li kev siv (Jiao li al., 2009). Kev ua haujlwm ntawm alkaline phosphatase (ALP) thiab qib Ca thiab P tau kwv yees los ntawm kev siv lub tshuab ntsuas tsis siv neeg (Ciba-Corning 550, USA).

Pob txha Mineral Density thiab Micro-Computed Tomography Analysis

Ob lub zog X-ray absorptiometry tshuab (Lunar, USA) tau ua haujlwm los kwv yees cov pob txha pob txha pob txha ntom ntom (BMD, g / cm2) ntawm txoj cai femur ntawm txhua tus nas nrog hom scan tau teeb tsa ua tsiaj me. Tom qab ntawd, tib lub femur tau siv los ntsuas cov duab 3D ntawm trabecular pob txha microarchitecture los ntawm kev siv micro-CT scanner apparatus (GE, American), thaj tsam ntawm kev txaus siab (ROI) tau xaiv los ntawm kev teeb tsa tib lub koom haum hauv lub phaj loj hlob ntawm femur ntawm txhua tus qauv, microarchitecture tsis suav nrog cov pob txha ntxhia cov ntsiab lus (BMC), cov ntaub so ntswg ntxhia (TMC), cov ntaub so ntswg ntxhia ntom ntom (TMD), trabecular sib cais (Tb. sp), trabecular tooj (Tb. N), thiab trabecular thickness ( Tb. Th) tau los ntawm kev txheeb xyuas ROI.

Western Blot Determination

Osteoclasts raug ntxias los ntawm kev siv RAW 264.7 hlwb ntxiv nrog MCSF (25 ng / ml) thiab RANKL (20 ng / ml) (Yang et al., 2019). Tom qab 6 hnub ntawm kev txhawb nqa ntev, cov hlwb osteoclast uas tau pom zoo rau kev ua haujlwm TRAP tau kho nrog lossis tsis muaj CDE (0.01, 0.1, thiab 1 mg / ml, raws li) rau 48 h, ces cov hlwb raug lysed los ntawm lysis tsis, thiab Cov supernatants tau centrifuged thiab sib cais los ntawm 10 feem pua ​​​​sodium dodecyl sulfate-polyacrylamide gel electrophoresis thiab xa mus rau PVDF daim nyias nyias. Lub daim nyias nyias tau thaiv nrog 5 feem pua ​​​​ntawm cov mis nyuj tsis rog ntawm qhov kub thiab txias rau 1 h thiab tom qab ntawd incubated nrog cov tshuaj tiv thaiv thawj zaug tiv thaiv TRAF6 (1:400), RANKL (1: 400), RANK (1: 400), IKK (1: 400). ), NFκBIA (1:400), OPG (1:400), PI3K (1:400), AKT (1:400), c-Fos (1:400), NFAT2 (1:400), -actin, thiab GAPDH (1:1,{44}}). Tib daim nyias nyias raug stripped thiab soj ntsuam dua nrog cov tshuaj tiv thaiv kab mob, kuaj pom los ntawm Image Lab Software thaum kawg. Txhua qhov kev sim tau rov ua dua peb zaug, nrog -actin lossis GAPDH raws li kev tswj hwm sab hauv.

Kev txheeb cais

Cov ntaub ntawv ntawm peb qhov kev sim, piav raws li qhov txhais tau tias ± SD, tau txheeb xyuas los ntawm kev siv ib txoj kev tsom xam ntawm qhov sib txawv raws li Dennett qhov kev sim (SPSS 22.0 software, SPSS, USA), p < {{3} }.05 raug suav hais tias yog qhov tseem ceeb.

cistanchekuj hu uasuab puamGinseng

TSEEM CEEB Main Chemical Constituents ofCistanches deserticola

Extract Los ntawm kev siv cov khoom siv coj los siv, xya feem ntau yog phenylethanoid glycosides tebchaw suav nrog echinacoside,cistanosideA, acteosideCov tshuaj isoacteoside,cistanosideC, 2'-acetylacteoside,thiab 6'-acetylacteosidetau txheeb xyuas uas twb muaj lawm nyob rau hauv no extract, lawv coj chromatography peaks thiab cov qauv tau qhia nyob rau hauv daim duab 1.

Cov teebmeem ntawmCistanches deserticolaExtract ntawm lub cev, Uterine, thiab qhov chaw mos qhov hnyav

Tsis muaj qhov sib txawv tseem ceeb hauv thawj qhov hnyav ntawm lub cev ua ntej kev phais. Txawm li cas los xij, 3 lub hlis tom qab kev ua haujlwm ntawm ovariectomized, lub cev hnyav ntawm cov nas hauv OVX cov qauv pawg tau nce 35.7 feem pua, thaum lub tsev menyuam thiab qhov chaw mos ntub dej hnyav tau poob qis los ntawm 91.2 thiab 61.8 feem pua, raws li piv rau kev dag ntxias. nas (p < 0.001).="" cde,="" nrog="" rau="" qhov="" ntau="" npaum="" li="" cas="" ntawm="" 200-800="" mg="" kg="" hnub,="" tsis="" pom="" muaj="" kev="" cuam="" tshuam="" rau="" cov="" qhov="" ceeb="" thawj="" ntawm="" lub="" cev="" hnyav="" thiab="" txo="" qhov="" hnyav="" ntawm="" lub="" tsev="" menyuam="" thiab="" qhov="" chaw="" mos,="" raws="" li="" daim="" duab="" 2="" qhia.="" txawm="" li="" cas="" los="" xij,="" sib="" txawv="" tag="" nrho="" rau="" cde,="" 1="" mg="" kg="" hnub="" ntawm="" ev="" tau="" nthuav="" tawm="" cov="" txiaj="" ntsig="" tseem="" ceeb="" estrogenic,="" uas="" tau="" txais="" lub="" cev="" hnyav="" thiab="" txo="" qis="" hauv="" lub="" cev="" thiab="" qhov="" chaw="" mos="" qhov="" hnyav="" hauv="" ovx="" nas="" tau="" thim="" rov="" qab="" los="" ntawm="" ev="">

Cov teebmeem ntawmCistanches deserticolaExtract ntawm pob txha Mineral ceev thiab pob txha Microarchitecture

Ib qho kev xav tsis txaus ntseeg tau pom nyob rau hauv OVX pawg nrog BMD yog {{0}}}.158 ± 0.016 g/cm2 piv rau cov nas sham nrog BMD yog {{10}}.180 ± 0.010 g/cm2 (p < 0.001),="" uas="" txhais="" tau="" tias="" bmd="" ntawm="" cov="" nas="" hauv="" ovx="" qauv="" pawg="" tau="" txo="" qis="" li="" 12.2="" feem="" pua="" ​​tom="" qab="" 12="" lub="" lis="" piam="" ntawm="" ovariectomized="" phais="" raws="" li="" piv="" rau="" cov="" sham="" nas.="" raws="" li="" daim="" duab="" 3="" pom,="" txhua="" tus="" cde="" kho="" nas="" pom="" qhov="" nce="" bmd="" los="" ntawm="" 15.1="" feem="" pua="" ​​(p=""><0.001), 8.1="" feem="" pua="" ​​(p=""><0.05), thiab="" 9.2="" feem="" pua="" ​​(p=""><0.05), feem,="" piv="" rau="" ovx="" qauv="" pawg.="" daim="" duab="" 3d="" thiab="" cov="" txiaj="" ntsig="" zoo="" sib="" xws="" ntawm="" trabecular="" pob="" txha="" microarchitecture="" ntawm="" nas="" tau="" pom="" hauv="" daim="" duab="" 4.="" raws="" li="" cov="" txiaj="" ntsig="" ntawm="" bmd,="" qhov="" pom="" tseeb="" txo="" qis="" ntawm="" thaj="" chaw="" trabecular="" tau="" txais="" hauv="" ovx="" qauv="" pawg="" piv="" nrog="" cov="" sham="" nas.="" txawm="" li="" cas="" los="" xij,="" qhov="" tsis="" zoo="" ntawm="" cov="" pob="" txha="" trabecular="" tau="" txhim="" kho="" ib="" nrab="" los="" ntawm="" cde="" kev="" cuam="" tshuam,="" tshwj="" tsis="" yog="" tmd="" thiab="" tb.="" th,="" lwm="" yam="" trabecular="" pob="" txha="" tsis="" muaj="" xws="" li="" bmc,="" tmc,="" thiab="" tb.="" n="" tau="" nce="" thiab="" tb.="" sp="" tau="" pom="" zoo="" txo="" ​​qis="" tom="" qab="" kev="" kho="">

Cov teebmeem ntawmCistanches deserticolaExtract ntawm Urine thiab Serum Biochemical Parameters

Kaum ob lub lis piam tom qab kev ua haujlwm ntawm ovariectomized, qhov poob qis tab sis tsis yog qhov ntsuas ntawm cov zis tso zis theem ntawm Ca tau pom nyob rau hauv OVX qauv pawg, raws li daim duab 5 tau pom, whereas theem ntawm cov zis P hauv nas ntawm OVX qauv pawg tau poob txog peb. lub sij hawm tsawg dua cov sham nas (p < 0.001).="" tom="" qab="" kev="" kho="" mob="" nrog="" cde="" (200–400="" mg="" kg="" hnub)="" rau="" 12="" lub="" lis="" piam,="" qhov="" txo="" qis="" ntawm="" cov="" ntshav="" ca="" thiab="" p="" tau="" raug="" tiv="" thaiv="" ntau="" heev="" (p=""><0.05), thiab="" txo="" qis="" tso="" zis="" theem="" ntawm="" p="" kuj="" inhibited="" nyob="" rau="" hauv="" tag="" nrho="" cov="" cde="" kho.="" pawg="" (p=""><>

Cov teebmeem ntawmCistanches deserticolaExtract ntawm pob txha tsim thiab Resorption Markers

Hais txog cov pob txha tsim cov cim ntawm ALP thiab BGP (Daim duab 6), cov haujlwm ntawm ALP, tsis yog BGP, tau txhim kho tau zoo nyob rau hauv tag nrho peb koob tshuaj CDE (200-800 mg / kg / hnub) kho pab pawg piv rau cov nas tsuag. Hais txog cov pob txha resorption cim ntawm TRAP, DPD, thiab cathepsin K (Daim duab 7), cov dej num ntawm tag nrho peb cov pob txha resorption

cov cim tau nce ntau ntxiv txog 20.9 ~ 74.8 feem pua ​​​​hauv cov nas ntawm OVX qauv pawg piv rau cov nas sham; thiab xav tias, CDE tau nthuav tawm lub peev xwm loj hauv kev tawm tsam tag nrho cov kev ntsuas no, uas cov haujlwm ntawm cathepsin K tau txo qis los ntawm 49.9 ~ 66.7 feem pua ​​(p < 0.001),="" qib="" ntawm="" dpd="" yog="" poob="" los="" ntawm="" 22.9="" ~="" 39.3="" feem="" pua="" ​​(p=""><0.01), thiab="" cov="" cuab="" yeej="" ntawm="" trap="" tau="" tiv="" thaiv="" los="" ntawm="" 20.1="" ~="" 27.6="" feem="" pua="" ​​(p=""><0.01), raws="" li="" piv="" nrog="" cov="" nas="" ntawm="" ovx="" qauv="">

Cov teebmeem ntawm Cistanches deserticola Extract ntawm Kev Ua Haujlwm ntawm Glutathione thiab Superoxide Dismutase as Well as Malondialdehyde Cov ntsiab lus

Tsis muaj qhov sib txawv tseem ceeb hauv kev ua ub no ntawm SOD thiab GSH ntawm OVX thiab sham pawg tom qab 3 lub hlis ntawm kev ua haujlwm ovariectomized, raws li daim duab 8 tau pom, thaum cov ntsiab lus ntawm MDA tau pom tseeb txhim kho hauv cov nas ntawm OVX.

cov qauv pab pawg los ntawm 68.2 feem pua ​​​​hauv kev sib piv rau cov sham nas, thiab cov qib MDA tau nce siab tau txo qis los ntawm qis, nruab nrab, thiab ntau npaum li cas ntawm CDE cov pab pawg kho mob nrog qhov inhibition feem pua ​​yog 66.1, 44.0, thiab 62.8 feem pua. , ntsig txog, piv nrog cov nas ntawm OVX qauv pawg.

Cov teebmeem ntawmCistanches deserticolaExtract ntawm Protein Expression Levels ntawm TRAF6, RANKL, RANK, NFκBIA, IKK, PI3K, AKT, OPG, c-Fos, thiab NFAT2

Raws li daim duab 9 tau pom, cov kab lus ntawm TRAF6, RANKL, RANK, IKK, NFκBIA, thiab NFAT2 tau txo qis-tswj thaum cov theem ntawm PI3K, AKT, OPG, thiab c-Fos tau pom tseeb tau tswj hwm tom qab kho nrog CDE piv rau. mus rau kev tswj. Hypothesized mechanism (Daim duab 10) uas CDE tuaj yeem txo qis- tswj cov qib ntawm RANKL, RANK, thiab TRAF6 thiab nce-tswj cov lus qhia ntawm OPG, yog li cov khoom sib txuas ntawm RANKL nrog RANK raug txo; Yog li ntawd, txoj hauv kev hauv qab no suav nrog NF-κB teeb liab tau raug tshem tawm thiab PI3K / AKT tau txhawb nqa, thiab cov kev taw qhia no ua ke ua rau kev ua kom cov c-Fos tau nce thiab NFAT2 tau inhibited, thiab thaum kawg cov pob txha osteoclastic resorption raug tiv thaiv. Kev sib tham