Oli e hloekileng ea Saposhnikovia divaricata bakeng sa kerese le sesepa e etsang wholesale diffuser oli ea bohlokoa e ncha bakeng sa li-diffuser tsa lehlaka.

tlhaloso e khuts'oane:

2.1. Tokisetso ea SDE

Li-rhizomes tsa SD li ile tsa rekoa e le setlama se omisitsoeng ho tloha Hanherb Co. (Guri, Korea). Lisebelisoa tsa semela li tiisitsoe ka mokhoa oa taxonomically ke Dr. Go-Ya Choi oa Korea Institute of Oriental Medicine (KIOM). Mohlala oa voucher (nomoro ea 2014 SDE-6) e kentsoe ho Korean Herbarium of Standard Herbal Resources. Li-rhizomes tse omisitsoeng tsa SD (320 g) li ile tsa ntšoa ka makhetlo a mabeli ka 70% ea ethanol (e nang le 2 h reflux) 'me mokelikeli o ile oa kenngoa tlas'a khatello e fokotsehileng. Decoction e ile ea tlhotloa, ea hloekisoa, 'me ea bolokoa mocheso oa 4 ° C. Litholoana tsa qotsulo e omisitsoeng ho tsoa linthong tse sa tsoakoang tse qalang e ne e le 48.13% (w/w).

2.2. Tlhahlobo ea Quantitative High-Performance Liquid Chromatography (HPLC).

Tlhahlobo ea Chromatographic e ile ea etsoa ka mokhoa oa HPLC (Waters Co., Milford, MA, USA) le mochine oa photodiode array detector. Bakeng sa tlhahlobo ea HPLC ea SDE, prim-O-glucosylcimifugin standard e ile ea rekoa ho Korea Promotion Institute for Traditional Medicine Industry (Gyeongsan, Korea), lesek-O-glucosylhamaudol le 4′-O-βD-glucosyl-5-O-methylvisamminol e ne e arotsoe ka har'a laboratori ea rona mme e khetholloa ka litlhahlobo tsa spectral, haholo-holo ke NMR le MS.

Mehlala ea SDE (0.1 mg) e ile ea qhibiliha ka 70% ethanol (10 mL). Karohano ea Chromatographic e entsoe ka kholomo ea XSelect HSS T3 C18 (4.6 × 250 mm, 5μm, Waters Co., Milford, MA, USA). Karolo ea selefouno e ne e e-na le acetonitrile (A) le 0.1% acetic acid ka metsing (B) ka tekanyo ea phallo ea 1.0 mL / min. Lenaneo la mekhahlelo e mengata le sebelisitsoe ka tsela e latelang: 5% A (0 mets), 5–20% A (0–10 min), 20% A (10–23 mets), le 20–65% A (23–40 min ). Leqhubu la ho lemoha le ile la hlahlojoa ho 210-400 nm mme la rekotoa ho 254 nm. Bophahamo ba ente e ne e le 10.0μL. Litharollo tse tloaelehileng bakeng sa qeto ea li-chromone tse tharo li ile tsa lokisoa boemong ba ho qetela ba 7.781 mg/mL (prim-O-glucosylcimifugin), 31.125 mg/mL (4′-O-βD-glucosyl-5-O-methylvisamminol), le 31.125 mg/mL (sek-O-glucosylhamaudol) ka methanol mme e bolokoa ho 4°C.

2.3. Tlhahlobo ea Ts'ebetso e Khahlanong le Ho ruruhaKa Vitro

2.3.1. Meetlo ea Lisele le Kalafo ea Mehlala

Lisele tsa RAW 264.7 li ile tsa fumanoa ho American Type Culture Collection (ATCC, Manassas, VA, USA) 'me tsa lengoa ka DMEM medium e nang le lithibela-mafu tsa 1% le 5.5% FBS. Lisele li ile tsa kenngoa moeeng o mongobo oa 5% CO2 ho 37°C. E le ho susumetsa lisele, setsi se ile sa nkeloa sebaka ke DMEM e ncha, le lipopolysaccharide (LPS, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) ho 1μg/mL e kentsoe boteng kapa bo le sieo ba SDE (200 kapa 400μg/mL) bakeng sa lihora tse ling tse 24.

2.3.2. Determination of Nitric Oxide (NO), Prostaglandin E2 (PGE2), Tumor Necrosis Factor-α(TNF-α), le Tlhahiso ea Interleukin-6 (IL-6).

Lisele li ne li phekoloa ka SDE 'me li tsosoa ka LPS bakeng sa 24 h. HA HO tlhahiso e ile ea hlahlojoa ka ho lekanya nitrite ho sebelisa reagent ea Griess ho latela thuto e fetileng [12]. Sephiri sa li-cytokine tsa ho ruruha PGE2, TNF-α, 'me IL-6 e ne e ikemiselitse ho sebelisa ELISA kit (R & D systems) ho latela litaelo tsa moetsi. Liphello tsa SDE ho NO le tlhahiso ea cytokine li ile tsa khethoa ho 540 nm kapa 450 nm ho sebelisa Wallac EnVision.™sebali sa microplate (PerkinElmer).

2.4. Tlhahlobo ea Mosebetsi oa AntiosteoarthritisHo Vivo

2.4.1. Liphoofolo

Litoeba tse tona tsa Sprague-Dawley (libeke tse 7) li ile tsa rekoa ho Samtako Inc. (Osan, Korea) 'me tsa lula tlas'a maemo a laoloang ka potoloho ea leseli / lefifi la 12-h.°C le% mongobo. Likhoto li ile tsa fuoa lijo tsa laboratori le metsiad libitum. Mekhoa eohle ea liteko e entsoe ho latela litataiso tsa Mekhatlo ea Sechaba ea Bophelo (NIH) le ho amoheloa ke Komiti ea Tlhokomelo ea Liphoofolo le Tšebeliso ea univesithi ea Daejeon (Daejeon, rephabliki ea Korea).

2.4.2. Ho kenngoa ha OA le MIA ho Likhoto

Liphoofolo li ne li sa tloaeleha 'me li abeloa lihlopha tsa phekolo pele ho qala thuto (ka sehlopha). Tharollo ea MIA (3 mg/50μL ea 0.9% saline) e ne e kenngoa ka ho toba sebakeng sa intra-articular ea lengole le letona tlas'a anesthesia e bakoang ke motsoako oa ketamine le xylazine. Likhoto li ne li arotsoe ka lihlopha tse 'nè: (1) sehlopha sa saline se se nang ente ea MIA, (2) sehlopha sa MIA se nang le ente ea MIA, (3) sehlopha sa phekolo ea SDE (200 mg / kg) ka ente ea MIA, le (4) ) sehlopha sa phekolo ea indomethacin- (IM-) (2 mg/kg) ka ente ea MIA. Likhoto li ne li tsamaisoa ka molomo le SDE le IM 1 beke pele ho ente ea MIA bakeng sa libeke tsa 4. Tekanyo ea SDE le IM e sebelisitsoeng thutong ena e ne e thehiloe ho ba hiriloe lithutong tse fetileng [10,13,14].

2.4.3. Litekanyo tsa Kabo ea Boima ba Hindpaw

Ka mor'a ho kenngoa ha OA, tekanyo ea pele ea bokhoni ba ho jara boima ba li-hindpaws e ile ea senyeha. Mohlahlobi oa ho hloka matla (Linton instrumentation, Norfolk, UK) o ne o sebelisetsoa ho hlahloba liphetoho ho mamello ea boima ba 'mele. Likhoto li ne li kenngoa ka hloko ka phaposing ea ho lekanya. Matla a ho jara boima a sebelisoang ke leoto le ka morao a ile a lekanngoa ka nako ea 3 s. Karo-karolelano ea boima ba 'mele e baloa ka equation e latelang: [boima bo ka letsohong le letona le ka morao/(boima ka letsohong le letona le ka morao + boima ka letsohong le letšehali)] × 100 [15].

2.4.4. Litekanyo tsa Maemo a Serum Cytokine

Li-sampuli tsa mali li ne li le centrifuged ho 1,500 g bakeng sa metsotso e 10 ho 4 ° C; joale serum ea bokelloa ebe e bolokoa ho −70°C ho fihlela e sebelisoa. Boemo ba IL-1β, IL-6, TNF-α, 'me PGE2 ka serum e ne e lekantsoe ho sebelisoa lisebelisoa tsa ELISA tse tsoang ho R&D Systems (Minneapolis, MN, USA) ho latela litaelo tsa moetsi.

2.4.5. Tlhahlobo ea Nako ea 'Nete ea Quantitative RT-PCR

Kakaretso ea RNA e ne e ntšitsoe liseleng tse kopantsoeng tsa lengole ho sebelisa TRI reagent® (Sigma-Aldrich, St. Louis, MO, USA), e fetoletsoeng ka morao ho cDNA le PCR-amplified ho sebelisa TM One Step RT PCR kit e nang le SYBR e tala (Applied Biosystems , Grand Island, NY, USA). PCR ea nako ea sebele e ile ea etsoa ho sebelisoa Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Grand Island, NY, USA). Tatelano ya mathomo le tatelano ya probe di bontshitswe ho Lethathamo1. Li-aliquots tsa sampole ea cDNAs le palo e lekanang ea GAPDH cDNA li ile tsa holisoa ka motsoako o moholo oa TaqMan® Universal PCR o nang le DNA polymerase ho latela litaelo tsa moetsi (Applied Biosystems, Foster, CA, USA). Maemo a PCR e ne e le 2 min ho 50 ° C, 10 min ho 94 ° C, 15 s ho 95 ° C, le 1 min ho 60 ° C bakeng sa lipotoloho tsa 40. Khokahano ea phatsa ea lefutso e ile ea khethoa ho sebelisoa mokhoa oa ho bapisa oa Ct (nomoro ea potoloho ea moeli pakeng tsa sebaka sa amplification le threshold), ho latela litaelo tsa moetsi.

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Lintlha tsa Sehlahisoa

Li-tag tsa Sehlahisoa

Osteoarthritis (OA) ke lefu la masapo le manonyeletso le atileng haholo ho batho ba tsofetseng.1]. OA ke boemo bo bakoang ke karolo e 'ngoe ea kotsi, tahlehelo ea sebopeho sa lefufuru le ts'ebetso, le ho se sebetse hantle ha litsela tsa ho ruruha le ho thibela ho ruruha [2,3]. E ama haholo-holo lefufuru le lesapo la subchondral la manonyeletso a synovial mme e fella ka ho hloleha ha manonyeletso, ho lebisang bohlokong ba boima ba 'mele ho kenyelletsa ho tsamaea le ho ema [4].

OA ha e na pheko, kaha ho thata haholo ho khutlisetsa lefufuru hang ha le senyehile [5]. Sepheo sa phekolo ke ho fokotsa bohloko, ho boloka kapa ho ntlafatsa ho tsamaea ha manonyeletso, ho eketsa matla a manonyeletso, le ho fokotsa liphello tse holofetseng tsa lefu lena. Kalafo ea Pharmacological ea OA e ikemiselitse ho fokotsa bohloko e le ho eketsa tšebetso ea manonyeletso ea mokuli le boleng ba bophelo. Le hoja timetso ea lefufuru e le ketsahalo e ka sehloohong ho OA, ho senyeha ha collagen ke ketsahalo ea bohlokoa e khethollang tsoelo-pele e ke keng ea khutlisoa ea OA hammoho le ho ruruha [6,7]. Liphekolo tse nang le ts'ebetso e khahlanong le ho ruruha le chondroprotective li lebeletsoe ho fokotsa bohloko le ho boloka botšepehi ba matrix ho bakuli ba OA.

Ka hona, ho fokotsa ho ruruha ho ka ba molemo tsamaisong ea OA. Liphuputso tsa morao-rao li fana ka maikutlo a mesebetsi ea tšireletso bakeng sa mehloli ea litlama tsoelo-peleng ea OA, mabapi le ho fokotsa ho ruruha ha chondrocyte le ho senyeha ho eketsehileng ha lefufuru, ka bokhoni ba bona ba ho sebelisana le lisele tse amanang le manonyeletso, ho fella ka ho fokotsa bohloko ba manonyeletso.8].

Motso oaSaposhnikovia divaricataSchischkin (Umbelliferae) e 'nile ea sebelisoa haholo meriana ea setso bakeng sa phekolo ea hlooho, bohloko, ho ruruha le ramatiki Korea le Chaena.9,10]. Liphello tse fapaneng tsa pharmacological tsaSaposhnikovia divaricata(SD) e boetse e kenyelletsa anti-inflammatory, analgesic, antipyretic, le antiarthritic properties [9,11]. Phuputso ea morao tjena e bonts'itse hore SD chromone extract e na le litlamorao tse ka bang teng tsa antirheumatoid arthritis mokhoeng oa toeba oa ramatiki e bakoang ke collagen.10]; leha ho le joalo, ho entsoe liphuputso tse fokolang ho tšehetsa tšebetso ea anti-inflammatory le antiarthritis eaSaposhnikovia divaricata(SDE)

Ka hona, thuto ea hona joale e ile ea batlisisa mesebetsi ea anti-inflammatory le antiosteoarthritis ea 70% ea ethanol extract ea SD. Ntlha ea pele, phello e khahlanong le ho ruruha ea SDE e ile ea hlahlojoaka vitroka liseleng tsa RAW 264.7 tse hlahisitsoeng ke LPS. Ka mor'a moo, phello ea antiosteoarthritis ea SDE e ile ea lekanngoa ka ho hlahloba kabo ea boima ba 'mele, ho senyeha ha lefufuru la articular, le likarabo tsa ho ruruha ka mokhoa oa rat oa monosodium iodoacetate- (MIA-) e entsoeng OA.