PART 1 Cistanche Tubulosa (Schenk) Wight Extract Enhances Hindlimb Performance Thiab Attenuates Myosin Heavy Chain IId/IIx Qhia hauv Cast-Immobilized nas

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

1Division of Neuromedical Science, Institute of Natural Medicine, University of Toyama, 2630 Sugitani, Toyama 930-0194, Nyiv 2Department of Japanese Oriental Medicine, Graduate School of Medicine thiab Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama {{5 }}, Nyiv

Kev sau ntawv yuav tsum tau hais rau Chihiro Tohda; chihiro@inm.u-toyama.ac.jp ib

Tau txais 14 Lub Rau Hli 2019; Hloov kho 13 Cuaj hlis 2019; Tau txais 17 Cuaj hlis 2019; Tshaj tawm rau lub Kaum Hlis 22, 2019

Academic Editor: Wen-yi Kang

Copyright © 2019 Yoshiyuki Kimbara et al. Qhov no yog ib tsab xov xwm qhib nkag tau muab faib raws li Creative Commons Attribution License, uas tso cai rau kev siv tsis txwv, kev faib tawm, thiab luam tawm hauv ib qho nruab nrab uas muab cov haujlwm qub raug suav hais tias yog.

Skeletal leeg atrophy tau ntsib nyob rau hauv ntau qhov chaw kho mob, tab sis kev kho tshuaj pharmacological tseem tsis tau tsim.Cistanche tubulosa(Schenk) Wight yog tshuaj ntsuab siv hauv cov tshuaj Japanese thiab suav tshuaj. Hauv kev tshawb fawb tam sim no, peb tau tshawb xyuas qhov cuam tshuam ntawmC. tubulosa extract(CTE) ntawm cov leeg atrophied hauv vivo. Peb kuj tau tshawb xyuas hindlimb cam immobilization nyob rau hauv nas thiab tsim ib hom tshiab ntawm hindlimb-immobilizing cam khwb cia, muaj xws li daim txhuam cev zoo li daim kab xev thiab ib lub raj yas nyias. Siv txoj kev no, 3 tawm ntawm 4 pawg nas (n 11 rau txhua pab pawg) raug cam khwb cia-immobilized nyob rau hauv hindlimbs thiab tswj CTE los yog tsheb rau 13 hnub. Cov txheej txheem sham tau ua nyob rau hauv cov nas ntawm pawg plaub uas lub tsheb tau tswj hwm. Tom qab ntawd, cov leeg triceps surae (TS) tau raug tshem tawm. Txhawm rau txheeb xyuas qhov cuam tshuam ntawm cov txheej txheem tshiab cam khwb cia thiab CTE kev tswj hwm ntawm cov leeg nqaij atrophy, peb tau soj ntsuam TS ntub qhov hnyav thiab myofiber cross-sectional area (CSA). Peb kuj tau txiav txim siab MyHC IId / IIx qhia qib los ntawm kev ua rau sab hnub poob, vim tias lawv qhov kev nce ntxiv yog qhov pom ntawm kev tsis siv cov leeg nqaij atrophy, qhia txog kev ua haujlwm qeeb-rau-ceev myofibber hom hloov. Tsis tas li ntawd, peb tau ua ob qhov kev sim ntawm hindlimb kev ua tau zoo. Txoj kev tshiab cam khwb cia immobilization tseem ceeb txo TS ntub qhov hnyav thiab myofiber CSA. Qhov no tau nrog los ntawm kev tsis zoo ntawm hindlimb muaj nuj nqi thiab nce hauv MyHC IId / IIx qhia. Kev tswj hwm CTE tsis hloov TS ntub qhov hnyav lossis myofiber CSA; Txawm li cas los xij, nws tau qhia txog kev hloov pauv ntawm qhov poob ntawm hindlimb muaj nuj nqi thiab kev tawm tsam ntawm qhov nce MyHC IId / IIx qhia hauv cov nas cam khwb cia-immobilized. Peb qhov tshiab hindlimb cam khwb cia immobilization txoj kev ua kom cov leeg nqaij atrophy. CTE tsis cuam tshuam rau cov leeg nqaij tab sis txwv kev hloov pauv los ntawm qeeb mus rau ceev myofiber hom hauv cov nas cam khwb cia-immobilized, ameliorating hindlimb muaj nuj nqi deterioration.

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

1. Taw qhia

Cov kab mob pathophysiological cuam tshuam nrog cov leeg pob txha atrophy suav nrog sarcopenia [1], cachexia [2], thiab glucocorticoid-induced skeletal leeg atrophy [3]. Skeletal nqaij atrophy kuj yog tshwm sim los ntawm kev siv lub sijhawm ntev [4], uas tuaj yeem yog vim lub txaj so [5], kev siv cov casts rau kev kho mob pob txha lossis ligament raug mob [6], lossis qhov chaw [7]. Txij li muaj ntau yam kab mob tuaj yeem ua rau tus mob no, nws cuam tshuam rau feem pua ​​​​ntawm cov pejxeem.

Kev tawm dag zog txaus thiab kev noj zaub mov zoo yog qhov tseem ceeb los kho cov leeg nqaij atrophy [8]. Tsis tas li ntawd, qee qhov kev tshawb fawb tau tshaj tawm tias cov tshuaj tiv thaiv tshuaj, suav nrog kev xaiv androgen receptor modulators, tib neeg monoclonal antibodies tiv thaiv myostatin, thiab xaiv ghrelin receptor agonists, tau txais txiaj ntsig [9]; Txawm li cas los xij, cov txheej txheem no tseem tsis tau tsim.

Cov leeg nqaij atrophic tau piav qhia saum toj no yog nrog kev hloov pauv hauv myofiber hom profile thiab txo cov leeg nqaij. Cov leeg nqaij mob tshwm sim los ntawm kev poob ntawm neural influence thiab mechanical unloading induces a slow-rau-ceast myofiber type shift, and a change in myosin heavy chain (MyHC) isoform profille [10], signaled by increment of MyHC IId/IIx qhia qib [11, 12] ib. Kev hloov pauv sai-rau-slow myofiber yog hloov pauv los ntawm kev tswj hwm glucocorticoid, cachexia, sepsis, thiab lwm yam [10].

Cistancheyog ib hom tshuaj herbal originating los ntawmCistanche deserticolaYC Ma,C. tubulosa(Schenk) Wight, los yog C. salsa (CA Mey) Beck, raws li Japanese pharmacopeia [13]. Nyob rau hauv ib txwm Japanese (Kampo) thiab Suav tshuaj, nws yog siv los kho "mob raum tsis ua hauj lwm syndrome," yam ntxwv ntawm infertility, systemic leeg tsis muaj zog (tshwj xeeb tshaj yog nyob rau hauv lub nraub qaum thiab qis ceg), thiab pob txha tsis muaj zog [14, 15]. Nws paub tias muaj ntau yam khoom siv, suav nrog acteoside, echinacoside, thiab isoacteoside [16]. Cov extract ntawm cov tshuaj ntsuab no, los yog nws cov khoom siv tshuaj, tau pom tias muaj cardio-, hepato-, thiab neuroprotective cuam tshuam; antiaging, antifatigue, vasodilatory, thiab aphrodisiac cuam tshuam; antioxidant thiab anti-inflammatory kev ua; thiab muaj peev xwm txhim kho kev nco thiab kev kawm [15, 16]. Txawm li cas los xij, qhov kev ua tau zoo ntawm cov tshuaj ntsuab no los tiv thaiv lossis kho cov kab mob ntawm cov leeg nqaij tsis tau lees paub. Yog li ntawd, hauv kev tshawb fawb tam sim no, peb tau tshawb xyuas qhov cuam tshuam ntawmC. tubulosaExtract (CTE) raws li kev kho mob rau cov leeg nqaij atrophy.

Txog qhov kawg no, peb siv hindlimb cam khwb cia immobilization los ua kom cov leeg nqaij atrophy hauv nas, tsom, tshwj xeeb, ntawm cov khoom siv hauv cov cam khwb cia. Ntau yam txheej txheem cam khwb cia tau tshaj tawm, suav nrog plaster ntawm Paris cam khwb cia [17], fiberglass cam khwb cia [18], polypropylene microcentrifuge hlab [19], txwv tsis pub cov khoom siv zoo li cov paj rwb thiab steel mesh [20], yas pipettes nrog proximal sponge padding [21], cam khwb cia ua los ntawm cov ntaub qhwv tsis-elastic daim kab xev thiab vinyl-coated steel hlau [22], xaim ntxig nrog cov hlau phaj [23], thiab siv cov tawv nqaij phais stapler [24]. Qhov zoo tshaj plaws cam khwb cia yuav tsum muaj tsawg kawg nkaus invasive thiab nontraumatic kom tsis txhob inflammatory kev hloov vim o tuaj yeem ua rau cov leeg nqaij atrophy [25]. Tsis tas li ntawd, qhov kev ntsuam xyuas hindlimb nrog cov cam khwb cia yuav tsum ua tau nyob rau theem kawg ntawm qhov kev sim. Tsis tas li ntawd, cov cam khwb cia yuav tsum yooj yim los tuav yam tsis muaj kev paub tshwj xeeb lossis kev txawj ntse; nws yuav tsum yog lub teeb, pheej yig, yooj yim rau kev yuav khoom, thiab yuav tsum tsis txhob xav tau cov cuab yeej tshwj xeeb rau kev thov thiab tshem tawm.

Cov qauv tsiaj sib txawv ntawm cov leeg nqaij nkim tau tshaj tawm. Hindlimb unloading tau siv ntau zaus [11, 12] tab sis muaj ntau yam teeb meem, nrog rau tus Tsov tus tw necrosis, uas tuaj yeem ua rau muaj kab mob hauv lub cev [25]; deviation ntawm lub cev kua mus rau sab caudal, ua rau muaj kev txawv txav hauv cov hlab ntsha hindlimb [26]; extraordinary microgravity ntawm lub hindlimb, uas tuaj yeem cuam tshuam rau cov protein qhia, suav nrog cov enzymes mitochondrial [27]. Tag nrho cov xwm txheej no tuaj yeem cuam tshuam rau cov leeg nqaij. Ib txoj kev sib txawv induces nqaij atrophy los ntawm depriving cov leeg ntawm neuronal inputs (piv txwv li, ntawm sciatic paj transection) [28]; txij li cov leeg yuav raug rau trophic yam los ntawm lub davhlau ya nyob twg neurons [29], txoj kev no, uas ua rau muaj qhov txawv txav ze-tag nrho cov txheej txheem denervated, tuaj yeem hloov cov txheej txheem ntawm cov leeg nqaij atrophy. Tsiaj txhu uas raug rau cov kab mob uas ua rau cov leeg nqaij atrophy, xws li ntshav qab zib mellitus [30], lub raum tsis ua haujlwm [31], kev tswj hwm corticosteroid [32], thiab kev laus lossis kev ua kom nrawm nrawm [33, 34], tuaj yeem siv los ua qauv ntawm tib neeg. kab mob; Txawm li cas los xij, cov kab mob no tuaj yeem cuam tshuam nrog cov txheej txheem ntawm cov leeg nqaij atrophy.

Yog li, peb xaiv hindlimb cam khwb cia immobilization los ua kom cov leeg nqaij atrophy, thiab peb nrhiav los tsim ib txoj hauv kev tshiab los ua nws hauv cov nas, uas peb tau tshawb xyuas qhov cuam tshuam ntawm CTE ntawm cov leeg nqaij atrophy.

CistancheKev noj zaub mov ntxivLub raumKab mobthiabBoostsLub zog

2. Cov ntaub ntawv thiab cov txheej txheem

2.1. Cog Khoom thiab Extraction.

CTE tau txais los ntawm Alps Pharmaceutical Industry Co., Ltd. (Hida, Gifu, Nyiv).

C. tubulosa cov stems tau sau los ntawm Shinjang Uyghur Aptonom Rayon, Cov Neeg Sawv Cev ntawm Tuam Tshoj. Tag nrho ntawm 193 kg ntawm cov hmoov sib xyaw ntawm C. tubulosa tau muab tso rau hauv 1,883 l ntawm 30 feem pua ​​​​ethanol. Qhov sib tov yog refluxed rau 2 teev ntawm 55-64 degree. Lub extract yield yog 7.2 feem pua ​​thiab muaj 3.36 feem pua ​​acteoside thiab 12.72 feem pua ​​echinacoside.

2.2. Cov nas.

Cov neeg laus ddY (12 lub lis piam) txiv neej nas (Nyiv SLC, Shizuoka, Nyiv) tau nyob nyias hauv lub tawb yas thiab khaws cia ntawm 12-h lub teeb / tsaus voj voog nyob rau hauv qhov ntsuas kub (25 ± 2 degree) vivarium. Txhua tus tsiaj tau tso cai rau kev nkag mus rau dej thiab chow (Labo MR Tshuag; Nosan Corporation, Yokohama, Kanagawa, Nyiv).

Txhua qhov kev sim tsiaj thiab cov txheej txheem tau ua raws li Cov Lus Qhia rau Kev Saib Xyuas thiab Kev Siv Cov Tsiaj Looj Meem ntawm Sugitani Campus ntawm University of Toyama. Tus lej pom zoo rau kev sim tsiaj yog A2016INM-3. Txhua qhov kev siv zog tau tsim los txo cov tsiaj txhu tsawg kawg nkaus.

2.3. Induction ntawm Hindlimb Muscle Atrophy thiab CTE Administration.

Cov nas tau muab faib ua 4 pawg (n 11 ib pab pawg): Hnub 2, txhua tus nas tau tshuaj loog, thiab lawv cov plaub hau raug tshem tawm ntawm tus ncej puab mus rau pob taws nrog ib tug depilatory cream (Epilat cream; Kracie, Tokyo, Nyiv). Tom ntej no, 3 tawm ntawm 4 pawg cam khwb cia (ntxiv rau) pawg raug cam khwb cia-immobilized raws li cov txheej txheem hauv qab no, raws li qhia hauv daim duab 1(a)–1(g):

Kauj Ruam 1. Txiav ib daim EPDM roj hmab ua npuas ncauj sealing daim kab xev (Cemedine Corporation, Tokyo, Nyiv) rau hauv ib tug 12 8 25 hli strip (Daim duab 1(a)).

Kauj Ruam 2. Tuav lub nraub qaum ntawm lub pob taws ntawm pob taws thiab muab nws maj mam muab qhwv rau ntawm pob luj taws nrog ib daim kab xev kaw (Daim duab 1(b)).

Kauj Ruam 3. Txiav ib lub mos PVC rwb thaiv tsev cap TIC-22 (Nichifu Corporation, Osaka, Nyiv), thiab npog lub sealing daim kab xev. Siv ob daim kab xev (PRO SELF handy cut ob-sided daim kab xev J-1300; Nitoms Inc., Tokyo, Nyiv) kom lawv ua raws (Daim duab 1(c)).

Kauj Ruam 4. Sau cov txiav kawg ntawm lub hau; txiav cov ntaub nplaum uas tsis muaj dej tsis zoo (huv daim kab xev; Asahipen Corporation, Osaka, Nyiv) mus rau kwv yees li. 4 48 mm loj thiab kho qhov sib sau ua ke (Daim duab 1(d)).

Kauj Ruam 5. Muab ib daim me me (kwv yees 5 30 hli) ntawm daim kab xev aluminium (AL-30; Nichiban Corporation, Tokyo, Nyiv) rau sab pem hauv ntej ntawm sab sauv kawg ntawm lub hau (Daim duab 1 (e)).

Kauj Ruam 6. Txiav daim kab xev yas (vinyl hluav taws xob rwb thaiv tsev daim kab xev no. 21; Nitto Denko Corporation, Osaka, Nyiv) mus rau kwv yees. 19 × 40 mm, thiab qhwv lub hau (Daim duab 1(f)).

Kauj Ruam 7. Ua tib yam txheej txheem ntawm qhov kev sib tshuam sab nraub qaum (Daim duab 1(g)).

Lub sij hawm immobilization siv nyob rau hauv txoj kev tshawb no yog 13 hnub, thiab cam khwb cia raug hloov yog hais tias ib los yog ntau tshaj ntawm cov nram qab no tej yam kev mob tshwm sim: hnyav pedal edema, hindlimb necrosis, loj ulcerations ntawm daim tawv nqaij, loosening ntawm cam khwb cia vim kev puas tsuaj, los yog cam khwb cia dislocation ntawm lub pob taws. Ntawm cov pab pawg neeg ntxiv, tsuas yog depilation tau ua (sham txheej txheem), thiab lub tsheb (lub cev saline) tau muab rau 13 hnub txij hnub tom qab depilation (hnub 2). Ntawm 3 pawg (ntxiv rau) pawg, 2 tau tswj hwm CTE (50 thiab 100 mg / kg / hnub; cam khwb cia (ntxiv ) / CTE -50 pawg thiab cam khwb cia (ntxiv rau) / CTE -100 pawg), thiab ib qho tau tswj hwm lub tsheb (ntaus (ntxiv rau) / pawg tsheb), kuj tseem los ntawm hnub 2. CTE tau yaj mus rau hauv physiological saline thiab muab txhua hnub ntawm qhov ncauj gavage rau 13 hnub.

2.4. Kev ntsuam xyuas ntawm Hindlimb Function thiab Hindlimb Muscle Strength.

Hnub 14, cov nas ntawm pawg cam khwb cia (ntxiv rau) tau tshuaj loog dua tom qab CTE tswj hwm, thiab cov casts raug tshem tawm. Hnub tom qab (hnub 15), tag nrho cov nas tau sim rau hindlimb muaj nuj nqi thiab cov leeg nqaij, siv ob qhov kev ntsuam xyuas hauv qab no:

2.4.1. Kev ntsuas ntawm Gait Ceev.

Ib lub 15 hli dav square ntoo platform tau tuav kab rov tav kwv yees li 50 cm ntawm hauv av. Tom qab ntawd tus nas tau muab tso rau ntawm lub platform, thiab kev nrawm nrawm tau soj ntsuam raws li tus tsiaj hla lub platform los ntawm kev kaw lus manually siv lub moos nres lub sijhawm taug kev deb ntawm 50 cm. Ua ntej, txhua tus nas tau nyob hauv lub platform los ntawm kev hla nws ob zaug raws li kev cob qhia.

2.4.2. Foot-Fault Test.

Txhawm rau ntsuas lub zog ntawm cov leeg hindlimb, peb tau ua qhov "ko taw-fault test," uas txhua tus nas tau muab tso rau ntawm lub platform dua, thiab tus naj npawb ntawm hindlimb slips los ntawm lub platform yog tabulated. Qhov kev sim no yog ib qho kev hloov kho ntawm qhov kev ntsuam xyuas pas nrig zoo li qub, nyob rau hauv cov pas nrig aerial tau kho nrog G-clamp, thiab cov nas raug tsim los hla lawv [35]. Kev ntsuam xyuas ko taw tau ua tiav peb zaug, thiab qhov nruab nrab ntawm cov footfalls hla peb ntu tau sau tseg. Muaj ib lub sijhawm so tsawg kawg yog 15 feeb ntawm kev sib tham.

2.5. Histological thiab Morphometric Analyzes ntawm Triceps Surae Muscle.

Tom qab qhov kev ntsuam xyuas ntawm hindlimb muaj nuj nqi, nas tau mob siab rau tshuaj loog thiab euthanatized los ntawm physiological saline perfusion. Tom ntej no, cov nqaij triceps surae (TS) tau excised los ntawm ob sab ntawm lub hindlimbs; Cov qauv ntawm sab laug hindlimb tau kho nyob rau hauv 4 feem pua ​​​​paraformaldehyde hauv PBS tov rau 24 h, tom qab ntawd hloov mus rau 30 feem pua ​​​​sucrose hauv PBS, thiab incubated rau cryoprotection thiab cryosectioning. Ua raws li qhov no, 12-μm cov ntu tuab tau raug txiav raws qhov me me ntawm thaj tsam nruab nrab plab ntawm txhua tus leeg siv lub cryostat (Leica 3050S; Leica systems, Nußloch, Lub teb chaws Yelemees). Xya mus rau kaum ntu ntu txuas ntxiv tau teeb tsa rau ntawm cov iav uas muaj tshuaj tiv thaiv kab mob (MAS-GP Hom A, Matsunami iav Kev Lag Luam, Osaka, Nyiv), tam sim ntawd qhuav siv lub tshuab ziab khaub ncaws, khaws cia ntawm 30 degree, thiab stained nrog hematoxylin-eosin staining (HE staining , ScyTek Laboratories, Logan, Utah, United States). Ib ntu tus neeg sawv cev rau ib tus tsiaj, uas qhov luv-axis hlais ntawm txhua tus myofiber tau pom meej meej, raug xaiv, thiab nws qhov chaw hla (CSA) raug ntsuas los ntawm Image J (National Institutes of Health, Bethesda, Maryland, United States) .

2.6. Western Blotting.

Tag nrho cov protein los ntawm txoj cai TS tau muab rho tawm los ntawm txoj kev hauv qab no. Txoj cai TS yog minced siv txiab thiab tam sim ntawd snap-kua hauv kua nitrogen, khaws cia ntawm 30 degree, thiab tom qab ntawd homogenized los ua cov ntaub so ntswg lysate. Thawed cov leeg tau incubated ice-txias rau 30 min nyob rau hauv 2 mL M-PER mammalian protein extract tov (Pierce Biotechnology, Rockford, Illinois, USA) thiab ces homogenized. Homogenates tau centrifuged ntawm 4 degree thiab 4000 g rau 10 min, thiab supernatants tau sau.

10 ug ntawm tag nrho cov protein nyob rau hauv txhua cov leeg nqaij lysate tau thauj khoom thiab sib cais los ntawm sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), tom qab ntawd los ntawm electrotransfer mus rau nitrocellulose membrane (0.2 μm, # 1620112, Bio-Rad Laboratories, Hercules, California, United States). Tsis muaj kev sib khi tshwj xeeb tau thaiv nrog 5 feem pua ​​​​cov mis tsis muaj roj qhuav (cov mis nyuj khov, 190-12865, FUJIFILM Wako Pure Chemical Industries, Osaka, Nyiv) hauv Tris-buffered saline nrog 0.05 feem pua ​​Tween 20 (TBS-T). Cov daim nyias nyias tau muab tso rau ib hmos ntawm 4 degree nrog thawj cov tshuaj tiv thaiv yaj hauv Can Get Signal immunoreaction enhancer solution 1 (Toyobo, Osaka, Nyiv). Tom qab agitating thiab yaug nrog TBS-T, cov membranes tau incubated rau 1 h nyob rau hauv chav tsev kub nrog horseradish peroxidase- (HRP-) conjugated thib ob antibody diluted nyob rau hauv Tau Txais Teeb Meem immunoreaction enhancer tov 2 (Toyobo, Osaka, Nyiv). Cov teeb liab ntawm daim nyias nyias tau kuaj pom los ntawm electrochemiluminescence siv luminol. Kev tsom xam Densitometric tau ua tiav siv CS Analyzer software (Atto, Tokyo, Nyiv). Cov tshuaj tiv thaiv raug xaiv thiab diluted raws li nram no: nas anti- MyHC IId/IIx monoclonal antibody (1 : 1000, clone A4.1025, #05-716, Merck Millipore, Burlington, Massachusetts, United States), nas anti-GAPDH loading tswj monoclonal antibody (1: 3500, G041 ABM, Vancouver, Canada), thiab HRP-conjugated tshis antimouse IgG poly- clonal secondary antibody H&L (1: 2000, Abcam, Cam- choj, United Kingdom).

2.7. Kev txheeb cais.

Tag nrho cov txiaj ntsig tau qhia raws li qhov ntsuas ± tus qauv yuam kev ntawm qhov nruab nrab (SEM). Qhov tseem ceeb ntawm cov kev sib piv ntawm cov pab pawg tau txheeb xyuas los ntawm ib-txoj kev ANOVA thiab tom qab hoc Dunnett qhov kev xeem lossis Tukey's HSD xeem; Kruskal–Wallis xeem thiab tom qab hoc Dunn qhov kev xeem; los yog ob txoj kev ANOVA nrog rov ntsuas dua thiab tom qab hoc Dunnett qhov kev xeem. Smirnov-Grubb qhov kev xeem tau siv los tshem tawm cov neeg sab nraud. Rau txhua qhov kev ntsuam xyuas kev txheeb cais, p qhov tseem ceeb tsawg dua 0.05 raug suav hais tias yog qhov tseem ceeb, thiab txhua qhov kev tshuaj ntsuam tau ua tiav siv GraphPad Prism 6 (GraphPad Software, La Jolla, California, United States).

CistancheKhoom noj khoom haus Anti-Aging thiab Boosts zog

3. Cov txiaj ntsig

3.1. Tsiaj Kev Noj Qab Haus Huv thiab Cov nyhuv ntawm CTE ntawm Lub Cev Qhov hnyav.

Nyob rau hauv qhov pom kev, lub hindlimbs ntawm cam khwb cia-immobilized nas yog tag nrho cov atrophic, piv nrog rau cov cam khwb cia ( ) / tsheb pawg nas. Cast immobilization ua rau poob qhov hnyav ntawm lub cev los ntawm qhov kawg ntawm kev kawm theem. Los ntawm qhov sib txawv, lub cev qhov hnyav hloov pauv tsis tseem ceeb hauv cov cam khwb cia ( ) / tsheb pawg nas. Ob pawg cam khwb cia (ntxiv rau) / CTE pawg pom tsis muaj qhov hloov pauv hauv lub cev hnyav thaum piv nrog cov cam khwb cia (ntxiv rau) / pawg tsheb (Daim duab 2).

Cov teeb meem hnyav nyob rau hauv lub hindlimbs ntawm immobilized nas, xws li daim tawv nqaij necrosis los yog gangrene, tsis tau pom thiab tsis yog tus cwj pwm txawv txav hauv vivarium. Txawm li cas los xij, cov teeb meem me me, suav nrog ko taw edema, daim tawv nqaij ulcerations, thiab rubor ntawm lub hindlimbs, tau pom nyob rau hauv tag nrho cov nas cast-immobilized (Daim duab 1(h)). Loosing thiab dislocation ntawm cov cam khwb cia, ua rau kev hloov cam khwb cia, kuj tau pom nyob rau hauv feem ntau.

3.2. Qhov cuam tshuam ntawm CTE ntawm Cov leeg nqaij thiab cov leeg nqaij ua haujlwm.

Cov cam khwb cia (ntxiv rau) / pawg tsheb tau nthuav tawm qhov tseem ceeb ntau ntxiv ntawm hindlimb slips hauv kev kuaj ko taw (Daim duab 3(a)), thiab kev nrawm nrawm nrawm dua li hauv cov cam khwb cia () / pawg tsheb (Daim duab 3(b) )).

Kev kho CTE tau ua rau muaj kev cuam tshuam ntawm koob tshuaj rau kev txo qis hindlimb slips. Hauv pawg cam khwb cia (ntxiv rau )/CTE-100, kev kho mob tau txo qis (p 0.0604) hindlimb slips thaum piv nrog cov cam khwb cia (ntxiv rau) / pawg tsheb (Daim duab 3(a)). Raws li kev khiav ceev, ob qho tib si cam khwb cia (ntxiv rau )/CTE-50 thiab cam khwb cia (ntxiv rau )/CTE-100 pawg tsis muaj qhov txawv txav hauv kev nrawm piv nrog cov cam khwb cia (ntxiv rau) / pawg tsheb, txawm hais tias muaj yog me ntsis, tsis tseem ceeb npaum li cas ntawm koob tshuaj raws li kev sib tw ceev ceev (Daim duab 3(b)).

3.3. Wet Weight of TS thiab Myofiber CSA.

Qhov hnyav ntub ntawm TS tau txo qis nrog cam khwb cia immobilization thaum piv nrog cov cam khwb cia () / pawg tsheb (Daim duab 4(a)). Kev tswj hwm CTE tsis cuam tshuam TS ntub dej.

Qhov nruab nrab myofiber CSA hauv txhua pab pawg kuj tau txheeb xyuas. Hauv pawg cam khwb cia (ntxiv rau) / pawg tsheb, pom muaj qhov txo qis hauv myofiber CSA piv nrog cov cam khwb cia ( ) / pawg tsheb (Daim duab 4(b)). Myofiber CSA tau muab faib ntxiv thiab tshuaj xyuas los ntawm kev faib tawm zaus, raws li pom hauv daim duab 4(c). Cov chav kawm CSA nquag tshaj plaws hauv pawg cam khwb cia ( ) / tsheb loj dua li hauv cov cam khwb cia (ntxiv rau) / pawg tsheb; Qhov feem pua ​​​​ntawm cov myofibers hauv 1350- 2250 μm2 CSA chav kawm yog, lossis zoo li, qhov tseem ceeb siab dua hauv pawg cam khwb cia ( ) / tsheb ntau dua li hauv pawg cam khwb cia (ntxiv rau ) / tsheb, thiab qhov sib txawv ntawm qhov pom tau pom hauv chav kawm CSA 0 900 μm2 pawg (Daim duab 4(c)). Kev faib tawm CSA tsis cuam tshuam los ntawm CTE kev tswj hwm hauv ob pawg casts (ntxiv rau) / CTE pawg, piv nrog cov cam khwb cia (ntxiv rau) / pawg tsheb (Daim duab 4(c)). Cov duab sawv cev myofiber muaj nyob hauv daim duab 4(d).

3.4. Qhov cuam tshuam ntawm CTE ntawm MyHC IId/IIx Kev Tshaj Tawm Qib hauv TS.

Western blotting tau ua nyob rau hauv 6 tus neeg sawv cev ntawm txhua pawg (Daim duab 5(a)). Cov qauv no tau raug xaiv los ntawm lawv qhov kev muab los ntawm cov tsiaj txhu uas ua pov thawj qhov kev sim ntsuas ko taw ua haujlwm ze tshaj plaws rau qhov txiaj ntsig ntawm lawv pawg.

Hauv pawg cam khwb cia (ntxiv rau) / pawg tsheb, MyHC IId / IIx tau nce ntau dua piv nrog pawg cast ( ) / tsheb. Qhov kev nce ntxiv no tau raug txwv los ntawm kev tswj hwm CTE. Hauv pawg cam khwb cia (ntxiv rau )/CTE-100 pawg, MyHC IId/IIx qhia tau qis dua piv nrog cov cam khwb cia (ntxiv rau) / pawg tsheb (p 0.0810) (Daim duab 5(b)) . GAPDH kev qhia, uas tau siv los ua kev tswj hwm sab hauv, tsis yog qhov txawv ntawm pawg (Daim duab 5(c)).

Cistanchetubulosa muaj ntau yam teebmeem