Saliida Saposhnikovia divaricata ee saafiga ah ee shumaca iyo saabuunta samaynta jumlo diffuser saliid muhiim ah oo cusub

Saliida Saposhnikovia divaricata ee saafiga ah ee shumaca iyo saabuunta samaynta jumlo qaybiye saliid muhiim ah

sharaxaad gaaban:

2.1. Diyaarinta SDE

Rhizomes-ka SD waxa laga soo gatay geedo qallalan oo laga keenay Hanherb Co. (Guri, Korea). Qalabka dhirta waxaa si canshuur ahaan u xaqiijiyay Dr. Go-Ya Choi oo ka tirsan Machadka Korea ee Daawada Oriental (KIOM). Muunada foojarka (lambarka 2014 SDE-6) ayaa lagu shubay Herbarium-ka Kuuriya ee Kheyraadka Dhirta ee caadiga ah. Rhizomes-ka qallalan ee SD (320 g) ayaa laba jeer laga soo saaray 70% ethanol (oo leh 2 h reflux) ka dibna soosaarku wuxuu ku ururay cadaadis yar. Decoction-ka waa la sifeeyay, lyophilized, waxaana lagu kaydiyay 4 ° C. Wax-soo-saarka soosaarka la qalajiyey ee alaabta bilowga ah wuxuu ahaa 48.13% (w/w).

2.2. Falanqaynta Dareeraha Waxqabadka Sare ee Tirada (HPLC).

Falanqaynta Chromatographic waxaa lagu sameeyay nidaamka HPLC (Waters Co., Milford, MA, USA) iyo qalab sawir qaade ah. Falanqaynta HPLC ee SDE, prim-OHalbeegga glucosylcimifugin waxaa laga soo iibsaday Machadka Horumarinta Kuuriya ee Warshadaha Daawada Dhaqanka (Gyeongsan, Kuuriya), iyosek-O-glucosylhamaudol iyo 4′-O-β-D-glucosyl-5-O-methylvisamminol ayaa lagu go'doomiyay shaybaadhkayaga waxaana lagu aqoonsaday falanqaynta muuqaalka ah, ugu horrayn NMR iyo MS.

Muunada SDE (0.1 mg) ayaa lagu milmay 70% ethanol (10 ml). Kala soocida Chromatographic waxaa lagu sameeyay tiir XSelect HSS T3 C18 (4.6 × 250 mm, 5).μm, Waters Co., Milford, MA, USA). Marxaladda guurguura waxay ka koobnayd acetonitrile (A) iyo 0.1% acetic acid ee biyaha (B) oo ah heerka socodka 1.0 mL/min. Barnaamij isjiid jiid badan ayaa loo isticmaalay sidan soo socota: 5% A (0 daqiiqo), 5–20% A (0–10 min), 20% A (10–23 min), iyo 20–65% A (23–40 min). ). Mawjada dhererka ogaanshaha waxaa lagu sawiray 210-400 nm waxaana lagu duubay 254 nm. Mugga duritaanku wuxuu ahaa 10.0μL. xalalka caadiga ah ee go'aaminta saddexda koromomoon ayaa la diyaariyey iyadoo la eegayo ugu dambeyntii 7.781 mg/mL (prim-O-glucosylcimifugin), 31.125 mg/mL (4′-O-β-D-glucosyl-5-O-methylvisamminol), iyo 31.125 mg/mL (sek-O-glucosylhamaudol) oo ku jira methanol waxaana lagu hayaa 4°C.

2.3. Qiimaynta Waxqabadka Ka-hortagga bararkaIn Vitro

2.3.1. Dhaqanka Unugga iyo Tusaalaha Daaweynta

Unugyada RAW 264.7 ayaa laga helay Ururinta Dhaqanka Nooca Maraykanka (ATCC, Manassas, VA, USA) waxayna ku koreen dhexdhexaad DMEM oo ka kooban 1% antibiyootiko iyo 5.5% FBS. Unugyada waxaa lagu dhex beeray jawi qoyan oo ah 5% CO2 at 37°C. Si loo kiciyo unugyada, dhexdhexaadinta waxaa lagu bedelay dhexdhexaad DMEM cusub, iyo lipopolysaccharide (LPS, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) at 1μg/mL ayaa lagu daray joogitaanka ama maqnaanshaha SDE (200 ama 400μg/ml) 24 saac oo dheeraad ah.

2.3.2. Go'aaminta Nitric Oxide (NO), Prostaglandin E2 (PGE2), Tumor Necrosis Factor-α(TNF-α), iyo Interleukin-6 (IL-6).

Unugyada waxaa lagu daweeyey SDE waxaana lagu kiciyey LPS 24 saac. NO wax soo saarka ayaa lagu falanqeeyay iyadoo la cabbirayo nitrite iyadoo la adeegsanayo Griess reagent sida lagu sheegay daraasad hore [12]. Soo saarida cytokines bararka PGE2, TNF-α, iyo IL-6 waxaa lagu go'aamiyay iyadoo la isticmaalayo qalab ELISA ah (R&D nidaamyada) sida waafaqsan tilmaamaha soo saaraha. Saamaynta SDE ee MAYA iyo wax soo saarka cytokine ayaa lagu go'aamiyay 540 nm ama 450 nm iyadoo la isticmaalayo Wallac EnVision™akhristaha microplate (PerkinElmer).

2.4. Qiimaynta Waxqabadka AntiosteoarthritisGudaha Vivo

2.4.1. Xoolaha

Jiirka Sprague-Dawley (7 toddobaad jir) ayaa laga soo iibiyay Samtako Inc. (Osan, Korea) waxaana lagu hoos jira xaalado la xakameeyey oo leh 12-saac iftiin / wareeg mugdi ah°C iyo% qoyaan. Jiirka ayaa la siiyay cunto shaybaar iyo biyoad libitum. Dhammaan hababka tijaabada ah waxaa loo sameeyay si waafaqsan tilmaamaha Machadka Qaranka ee Caafimaadka (NIH) oo ay ansixiyeen Guddiga Daryeelka Xoolaha iyo Isticmaalka ee Jaamacadda Daejeon (Daejeon, Jamhuuriyadda Kuuriya).

2.4.2. Soo saarista OA ee MIA ee jiirka

Xayawaanka ayaa la kala soocay waxaana loo qoondeeyay kooxaha daaweynta ka hor intaan la bilaabin daraasadda (koox walba). Xalka MIA (3 mg/50μL of 0.9% saline) ayaa si toos ah loogu duray booska gudaha ee jilibka midig ee suuxdinta oo ay keentay isku dhafka ketamine iyo xylazine. Jiirka ayaa si aan kala sooc lahayn loo kala qaybiyay afar kooxood: (1) kooxda cusbada leh ee aan lahayn duritaanka MIA, (2) kooxda MIA ee lagu duro MIA, (3) kooxda SDE lagu daweeyay (200 mg/kg) oo lagu duray MIA, iyo (4) ) indomethacin- (IM-) kooxda la daweeyay (2 mg/kg) oo lagu duray MIA. Jiirka waxaa lagu maamuli jiray afka SDE iyo IM 1 usbuuc ka hor duritaanka MIA muddo 4 toddobaad ah. Qiyaasta SDE iyo IM ee loo adeegsaday daraasaddan waxay ku saleysan tahay kuwa ka shaqeeya daraasadihii hore [10,13,14].

2.4.3. Cabbiraadaha Qaybinta Culayska Hindpaw

Ka dib soo saarista OA, dheelitirka asalka ah ee awoodda qaadista dhabarka ayaa la carqaladeeyey. Tijaabiye karti-darro (Linton instrumentation, Norfolk, UK) ayaa loo isticmaalay si loo qiimeeyo isbeddellada dulqaadka-qaadista. Jiirka ayaa si taxadar leh loo geliyey qolka cabbiraadda. Awoodda miisaan-qaadista ee ay samaysay addinka dambe waxa la isku celceliyey muddo 3 sc ah. Saamiga qaybinta miisaanka waxaa lagu xisaabiyay isla'egta soo socota: [miisaanka addinka dambe ee midig/(miisaanka addinka dambe ee midig + miisaanka dhabarka dambe ee bidix)] × 100 [15].

2.4.4. Cabbiraadaha Heerarka Serum Cytokine

Shaybaarada dhiiga waxa lagu kala saaray 1,500 g 10 min at 4°C; ka dibna serum-ka ayaa la ururiyay oo lagu kaydiyay -70 ° C ilaa isticmaalka. Heerarka IL-1β, IL-6, TNF-α, iyo PGE2 ee serumka ayaa lagu cabbiray iyadoo la isticmaalayo qalabyada ELISA ee R&D Systems (Minneapolis, MN, USA) sida ku cad tilmaamaha soo saaraha.

2.4.5. Falanqaynta Tirooyinka RT-PCR-waqtiga-dhabta ah

Wadarta guud ee RNA waxaa laga soo saaray unug wadajir ah oo jilibka ah iyadoo la adeegsanayo TRI reagent® (Sigma-Aldrich, St. Louis, MO, USA), oo loo rogay cDNA iyo PCR-la xoojiyay iyadoo la adeegsanayo xirmada TM One Step RT PCR oo leh cagaar SYBR , Grand Island, NY, USA). PCR-ta-waqtiga-dhabta ah ee tirade waxa lasameeyay iyadoo la isticmaalayo Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Grand Island, NY, USA). Taxanaha asaasiga ah iyo isku xigxiga baaritaanka ayaa lagu muujiyay shaxda1. Aliquots ee muunada cDNA-yada iyo xaddi siman oo GAPDH cDNA ah ayaa lagu kordhiyay TaqMan® Universal PCR isku dhafka sayid ee ka kooban DNA polymerase sida waafaqsan tilmaamaha soo saaraha (Applied Biosystems, Foster, CA, USA). Xaaladaha PCR waxay ahaayeen 2 min at 50°C, 10 min at 94°C, 15s at 95°C, iyo 1 min at 60°C ee wareegyada 40 Isku-duubnaanta hidde-sidaha bartilmaameedka ayaa lagu go'aamiyay iyadoo la adeegsanayo isbarbardhigga Ct (lambarka wareegga marinka ee barta-is-goysyada u dhexeeya goobta cod-weyneysiinta iyo marinka) habka, marka loo eego tilmaamaha soo-saaraha.

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Osteoarthritis (OA) waa xanuunka murqaha ee ugu badan iyo xanuunka wadajirka ah ee ugu badan ee dadka waayeelka ah.1]. OA waa xaalad ay sababto qayb ahaan dhaawac, luminta qaab-dhismeedka carjawda iyo shaqada, iyo habacsanaanta dariiqyada proinflammatory iyo anti-bararka.2,3]. Waxay ugu horreyntii saameysaa carjawda articular iyo lafaha subchondral ee kala-goysyada synovial waxayna keentaa guul-darro wadajir ah, taasoo keenta xanuunka miisaanka-qaadista oo ay ku jiraan socodka iyo istaagida [4].

Ma jirto wax daawo ah oo OA ah, maadaama ay aad u adag tahay in dib loo soo celiyo carjawda marka ay burburto [5]. Hadafka daawaynta waa in la yareeyo xanuunka, la ilaaliyo ama la hagaajiyo dhaqdhaqaaqa wadajirka ah, la kordhiyo xoogga kala-goysyada, iyo in la yareeyo saameynta naafada ee cudurka. Daawaynta dawooyinka ee OA waxay ujeedadeedu tahay in la yareeyo xanuunka si loo kordhiyo shaqada wadajirka ah ee bukaanka iyo tayada nolosha. Inkasta oo burburinta carjawda ay tahay dhacdada ugu weyn ee OA, hoos u dhaca kolajka ayaa ah dhacdada aasaasiga ah ee go'aamisa horumarka aan la bedeli karin ee OA ee la xidhiidha caabuqa.6,7]. Daawaynta ka hortagga bararka iyo dhaqdhaqaaqa chondroprotective ayaa la filayaa inay yareeyaan xanuunka oo ay ilaaliyaan sharafta matrixka ee bukaanka OA.

Sidaa darteed, hoos u dhigista bararka waxay u badan tahay inay faa'iido u leedahay maaraynta OA. Daraasadihii ugu dambeeyay waxay soo jeedinayaan doorarka ilaalinta khayraadka dhirta ee horumarka OA, marka la eego yaraynta caabuqa chondrocyte iyo burburinta carjawda dheeraadka ah, iyada oo loo marayo awooddooda ay ku dhexgalaan unugyada wadajirka ah, taas oo keentay in la yareeyo xanuunka wadajirka ah.8].

XididkaSaposhnikovia divaricataSchischkin (Umbelliferae) ayaa si weyn loogu isticmaalay daawada dhaqameed ee daaweynta madax-xanuun, xanuunka, bararka, iyo arthritis-ka ee Kuuriya iyo Shiinaha.9,10]. Saamaynta dawooyinka kala duwan eeSaposhnikovia divaricata(SD) waxa kale oo ka mid ah ka-hortagga bararka, xanuunka xanuunka, antipyretic, iyo sifooyinka antiarthritic.9,11]. Daraasad dhowaan la sameeyay ayaa muujisay in soosaarka koromomoonka SD uu leeyahay saameyn suurtagal ah antirheumatoid arthritis-ka ee moodeelka jiirka ee arthritis-ku keeno10]; si kastaba ha ahaatee, daraasado yar ayaa la sameeyay si loo taageero hawlaha ka hortagga bararka iyo ka hortagga arthritis-kaSaposhnikovia divaricatasoosaar (SDE).

Sidaa darteed, daraasadda hadda jirta waxay baaris ku samaysay hawlaha anti-bararka iyo antiosteoarthritis ee 70% ethanol ee SD. Marka hore, saameynta anti-bararka ee SDE ayaa la qiimeeyayin vitrounugyada RAW 264.7 ee ay keentay LPS. Marka xigta, saameynta antiosteoarthritis ee SDE waxaa lagu qiyaasay iyadoo la qiimeynayo qaybinta culeyska, hoos u dhaca carjawda articular, iyo jawaabaha bararka ee qaabka jiirka ee monosodium iodoacetate- (MIA-) ee OA.