Yakachena Saposhnikovia divaricata mafuta ekenduru uye sipo kugadzira wholesale diffuser yakakosha mafuta matsva ekupisa tsanga diffuser.

tsananguro pfupi:

 

2.1. Kugadzirira kweSDE

Iyo rhizomes yeSD yakatengwa semuriwo wakaomeswa kubva kuHanherb Co. (Guri, Korea). Zvigadzirwa zvemiti zvakasimbiswa taxonomically naDr. Go-Ya Choi weKorea Institute of Oriental Medicine (KIOM). Muenzaniso wevhocha (nhamba 2014 SDE-6) yakaiswa muKorea Herbarium yeStandard Herbal Resources. Dried rhizomes yeSD (320 g) yakatorwa kaviri ne 70% ethanol (ine 2 h reflux) uye iyo yakatorwa yakabva yaiswa pasi pekumanikidzwa kwakaderedzwa. Iyo decoction yakasefa, yakanyungudutswa, uye yakachengetwa pa4 ° C. Kukohwa kweakaomeswa akaomeswa kubva kune zvisina kusimba ekutanga zvinhu aive 48.13% (w/w).

 

2.2. Quantitative High-Performance Liquid Chromatography (HPLC) Analysis

Chromatographic analysis yakaitwa neHPLC system (Waters Co., Milford, MA, USA) uye photodiode array detector. Kune iyo HPLC kuongororwa kweSDE, iyo yekutanga-O-glucosylcimifugin standard yakatengwa kubva kuKorea Promotion Institute yeTraditional Medicine Industry (Gyeongsan, Korea), uyesec-O-glucosylhamaudol uye 4′-O-βD-glucosyl-5-O-methylvisamminol yakaparadzaniswa mukati merabhoritari yedu uye yakaonekwa ne spectral analyses, kunyanya neNMR uye MS.

SDE samples (0.1 mg) yakanyungudika mu70% ethanol (10 mL). Kupatsanurwa kweChromatographic kwakaitwa neXSelect HSS T3 C18 column (4.6 × 250 mm, 5μm, Waters Co., Milford, MA, USA). Chikamu chekufambisa chaive neacetonitrile (A) uye 0.1% acetic acid mumvura (B) pamwero wekuyerera we 1.0 mL / min. A multistep gradient program yakashandiswa sezvinotevera: 5% A (0 min), 5–20% A (0–10 min), 20% A (10–23 min), uye 20–65% A (23–40 min. ) Iyo yekuona wavelength yakaongororwa pa210-400 nm uye yakanyorwa pa254 nm. Iro jekiseni vhoriyamu yaive 10.0μL. Yakajairwa mhinduro yekugadzwa kwemakromone matatu yakagadzirirwa pachikamu chekupedzisira che7.781 mg/mL (prim-O-glucosylcimifugin), 31.125 mg/mL (4′-O-βD-glucosyl-5-O-methylvisamminol), uye 31.125 mg/mL (sec-O-glucosylhamaudol) mumethanol uye yakachengetwa pa4°C.

2.3. Kuongororwa kweAnti-Inflammatory ActivityIn Vitro

2.3.1. Cell Culture uye Sample Treatment

RAW 264.7 masero akawanikwa kubva kuAmerican Type Culture Collection (ATCC, Manassas, VA, USA) uye akakurira muDMEM medium ine 1% antibiotics uye 5.5% FBS. Masero akaiswa mumhepo ine humidified ye5% CO2 pa37°C. Kukurudzira masero, iyo yepakati yakatsiviwa neDMEM mitsva yepakati, uye lipopolysaccharide (LPS, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) pa1μg/mL yakawedzerwa mukuvapo kana kusavapo kweSDE (200 kana 400μg/mL) kwemamwe 24 h.

2.3.2. Kutsunga kweNitric Oxide (NO), Prostaglandin E2 (PGE2), Tumor Necrosis Factor-α(TNF-α), uye Interleukin-6 (IL-6) Kugadzirwa

Masero akabatwa neSDE uye akakurudzirwa neLPS ye24 h. HAPANA kugadzirwa kwakaongororwa nekuyera nitrite uchishandisa iyo Griess reagent maererano neongororo yapfuura [12]. Secretion yezvinoputika cytokines PGE2, TNF-α, uye IL-6 yakatemerwa kushandisa ELISA kit (R&D masisitimu) zvinoenderana nemirairo yemugadziri. Mhedzisiro yeSDE paNO uye cytokine kugadzirwa kwakatemwa pa540 nm kana 450 nm uchishandisa Wallac EnVision.™microplate reader (PerkinElmer).

2.4. Kuongororwa kweAntiosteoarthritis ActivityMuVivo

2.4.1. Mhuka

Male Sprague-Dawley rats (7 mavhiki ekuberekwa) akatengwa kubva kuSamtako Inc. (Osan, Korea) uye akachengetwa pasi pemamiriro ezvinhu akadzora ane 12-h chiedza / rima kutenderera pa.°C uye% humidity. Makonzo akapiwa chikafu chemurabhoritari nemvuraad libitum. Maitiro ese ekuyedza akaitwa achiteerana neNational Institutes of Health (NIH) nhungamiro uye yakatenderwa neAnimal Care uye Use Committee yeyunivhesiti yeDaejeon (Daejeon, republic yeKorea).

2.4.2. Induction yeOA ine MIA mumakonzo

Mhuka dzacho dzaive dzakarongeka uye dzakagoverwa kumapoka ekurapa kusati kwatanga chidzidzo (paboka). MIA mhinduro (3 mg/50μL ye0.9% saline) yakanyatsoiswa mu intra-articular space yemabvi orudyi pasi peanesthesia inokonzerwa nemusanganiswa we ketamine uye xylazine. Makonzo akapatsanurwa zvisina tsarukano mumapoka mana: (1) boka resaline risina jekiseni reMIA, (2) boka reMIA rine jekiseni reMIA, (3) boka rinorapwa SDE (200 mg/kg) ine jekiseni reMIA, uye (4) ) boka rinorapwa indomethacin- (IM-) (2 mg/kg) ine jekiseni reMIA. Makonzo aipihwa nemuromo neSDE uye IM 1 vhiki pamberi peMIA jekiseni kwemavhiki e4. Muyero weSDE uye IM wakashandiswa muchidzidzo ichi waive wakavakirwa pane vakamboshanda muzvidzidzo zvekare [10,13,14].

2.4.3. Zviyero zveHindpaw Weight-Bearing Distribution

Mushure meOA induction, chiyero chepakutanga mukuremerwa kwekugona kwehindpaws chakakanganiswa. An incapacitance tester (Linton instrumentation, Norfolk, UK) yakashandiswa kuongorora shanduko mukuremerwa kwekutakura. Makonzo ainyatsoiswa mukamuri rokuyeresa. Simba rekusimudza huremu rinoitwa negumbo rekumashure rakaitwa pakati penguva ye3 s. Huremu hwekugovanisa huremu hwakaverengerwa neinotevera equation: [huremu kuruoko rwerudyi / (huremu kuruoko rwerudyi + uremu kuruoko rweruboshwe)] × 100 [15].

2.4.4. Zviyero zveSerum Cytokine Levels

Zviyero zveropa zvaive centrifuged pa 1,500 g ye 10 min pa 4 ° C; zvino serum yakaunganidzwa uye yakachengetwa pa −70 ° C kusvikira yashandiswa. Iwo mazinga eIL-1β, IL-6, TNF-α, uye PGE2 muserum yakayerwa uchishandisa ELISA kits kubva kuR&D Systems (Minneapolis, MN, USA) maererano nemirairo yemugadziri.

2.4.5. Real-Nguva Quantitative RT-PCR Analysis

Yese RNA yakatorwa kubva mumabvi akabatanidzwa tishu uchishandisa TRI reagent® (Sigma-Aldrich, St. Louis, MO, USA), reverse-yakanyorwa mu cDNA uye PCR-yakasimudzwa uchishandisa TM Imwe Nhanho RT PCR kit ine SYBR yegirini (Applied Biosystems. , Grand Island, NY, USA). Real-time quantitative PCR yakaitwa pachishandiswa Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Grand Island, NY, USA). The primer sequences uye probe-sequence inoratidzwa muTable1. Aliquots yemasampuli cDNAs uye yakaenzana huwandu hweGAPDH cDNA yakakwidziridzwa neTaqMan® Universal PCR master musanganiswa ine DNA polymerase zvinoenderana nemirairo yemugadziri (Applied Biosystems, Foster, CA, USA). PCR mamiriro aiva 2 min pa 50 ° C, 10 min pa 94 ​​° C, 15 s pa 95 ° C, uye 1 min pa 60 ° C ye 40 mitsara. Iko kusungirirwa kwejini rinotarirwa kwakatemwa uchishandisa iyo yekuenzanisa Ct (chikumbaridzo kutenderera nhamba pamuchinjiko-pakati pakati pekukudza chidziro uye chikumbaridzo) nzira, zvinoenderana nemirairo yemugadziri.

FOB Mutengo:US $0.5 - 9,999 / Piece

Min.Order Quantity:100 Piece/Zvimedu

Supply Ability:10000 Piece / Zvimedu pamwedzi