ʻO ka aila Saposhnikovia divaricata maʻemaʻe no ke kukui a me ke kopa e hana ana i ka diffuser kūʻai nui i ka aila nui hou no nā diffusers kuni ʻohe.

ʻO ka aila Saposhnikovia divaricata maʻemaʻe no ke kukui a me ke kopa e hana ana i nā diffuser kūʻai nui i ka aila nui hou no nā diffusers puhi ʻohe.

wehewehe pōkole:

2.1. Ka hoomakaukau ana o SDE

Ua kūʻai ʻia nā rhizomes o SD ma ke ʻano he lāʻau maloʻo mai Hanherb Co. (Guri, Korea). Ua hōʻoia ʻia nā mea kanu e Kauka Go-Ya Choi o ka Korea Institute of Oriental Medicine (KIOM). Ua waiho ʻia kahi palapala hōʻailona (helu 2014 SDE-6) i ka Korean Herbarium of Standard Herbal Resources. Ua unuhi ʻia nā rhizomes maloʻo o SD (320 g) i ʻelua me ka 70% ethanol (me ka 2 h reflux) a laila hoʻopaʻa ʻia ka extract ma lalo o ke kaomi hoʻemi. Hoʻopili ʻia ka decoction, lyophilized, a mālama ʻia ma 4 ° C. ʻO 48.13% (w/w) ka loaʻa ʻana mai o nā mea hoʻomaloʻo maloʻo.

2.2. Kūlana Kiʻekiʻe-Hiʻona Liquid Chromatography (HPLC).

Hana ʻia ka nānā ʻana Chromatographic me kahi ʻōnaehana HPLC (Waters Co., Milford, MA, USA) a me kahi mea ʻike kiʻi kiʻi kiʻi. No ka hōʻike HPLC o SDE, ʻo ka prim-O-glucosylcimifugin standard i kūʻai ʻia mai ka Korea Promotion Institute for Traditional Medicine Industry (Gyeongsan, Korea), asec-O-glucosylhamaudol a me 4′-O-β-D-glucosyl-5-OUa hoʻokaʻawale ʻia -methylvisamminol i loko o kā mākou hale hana a ʻike ʻia e nā loiloi spectral, ma mua o NMR a me MS.

Ua hoʻoheheʻe ʻia nā laʻana SDE (0.1 mg) i 70% ethanol (10 mL). Hana ʻia ka hoʻokaʻawale Chromatographic me kahi kolamu XSelect HSS T3 C18 (4.6 × 250 mm, 5μm, Waters Co., Milford, MA, USA). ʻO ka māhele kelepona he acetonitrile (A) a me ka 0.1% acetic acid i loko o ka wai (B) ma kahi kahe kahe o 1.0 mL / min. Ua hoʻohana ʻia kahi papahana gradient multistep penei: 5% A (0 min), 5–20% A (0–10 min), 20% A (10–23 min), a me 20–65% A (23–40 min). ). Ua nānā ʻia ka lōʻihi hawewe ʻike ma 210-400 nm a hoʻopaʻa ʻia ma 254 nm. ʻO ka nui o ka injection he 10.0μL. Ua hoʻomākaukau ʻia nā haʻina maʻamau no ka hoʻoholo ʻana i nā chromones ʻekolu i ka hopena hope o 7.781 mg/mL (prim-O-glucosylcimifugin), 31.125 mg/mL (4′-O-β-D-glucosyl-5-O-methylvisamminol), a me 31.125 mg/mL (sec-O-glucosylhamaudol) i loko o ka methanol a mālama ʻia ma 4°C.

2.3. Ka Loiloi o ka Hana Anai-InflammatoryIn Vitro

2.3.1. Ka Moʻomeheu Cell a me ka Laʻana Lapaʻau

Ua loaʻa nā pūnaewele RAW 264.7 mai ka American Type Culture Collection (ATCC, Manassas, VA, USA) a ua ulu i loko o ka medium DMEM i loaʻa ka 1% antibiotic a me 5.5% FBS. Hoʻokomo ʻia nā kelepona i loko o kahi ea humidified o 5% CO2 ma 37 ° C. No ka hoʻouluʻana i nā pūnaewele, ua hoʻololiʻia ka medium me ka DMEM hou, a me ka lipopolysaccharide (LPS, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) ma 1.μUa hoʻohui ʻia ka g/mL ma ke alo a i ʻole ka nele o SDE (200 a i ʻole 400μg/mL) no 24 mau hola hou.

2.3.2. Ka Hoʻoholo ʻana o ka Nitric Oxide (NO), Prostaglandin E2 (PGE2), Tumor Necrosis Factor-α(TNF-α), a me Interleukin-6 (IL-6) Production

Ua mālama ʻia nā kelepona me SDE a hoʻoulu ʻia me LPS no 24 mau hola. NO ka hana ʻana i kālailai ʻia ma ke ana ʻana i ka nitrite me ka hoʻohana ʻana i ka Griess reagent e like me kahi noiʻi mua.12]. Ka huna o na cytokines inflammatory PGE2, TNF-α, a ua hoʻoholo ʻia ʻo IL-6 me ka hoʻohana ʻana i kahi kit ELISA (R&D system) e like me nā ʻōlelo a ka mea hana. Ua hoʻoholo ʻia nā hopena o SDE ma NO a me ka hana cytokine ma 540 nm a i ʻole 450 nm me ka hoʻohana ʻana i kahi Wallac EnVision.™mea heluhelu microplate (PerkinElmer).

2.4. Ka Loiloi o ka Hana AntiosteoarthritisMa Vivo

2.4.1. Holoholona

Ua kūʻai ʻia nā ʻiole kāne Sprague-Dawley (7 mau pule) mai Samtako Inc. (Osan, Korea) a mālama ʻia ma lalo o nā kūlana i hoʻomalu ʻia me ka pōʻaiapuni māmā/ʻeleʻele 12-h ma°C a% haʻahaʻa. Hāʻawi ʻia nā ʻiole i kahi meaʻai a me ka waiad libitum. Ua hana ʻia nā kaʻina hana hoʻokolohua a pau e like me nā alakaʻi alakaʻi o nā National Institutes of Health (NIH) a ʻae ʻia e ke Kōmike mālama holoholona a me ka hoʻohana ʻana o ke kulanui ʻo Daejeon (Daejeon, repubalika o Korea).

2.4.2. Hoʻokomo ʻia o OA me MIA ma nā ʻiole

Hoʻonohonoho ʻia nā holoholona a hāʻawi ʻia i nā hui lapaʻau ma mua o ka hoʻomaka ʻana o ke aʻo ʻana (no kēlā me kēia hui). Hoʻonā MIA (3 mg/50μL o 0.9% saline) ua hoʻokomo pololei ʻia i loko o ka intra-articular space o ka kuli ʻākau ma lalo o ka anesthesia i hoʻokomo ʻia me ka hui ʻana o ketamine a me xylazine. Ua hoʻokaʻawale ʻia nā ʻiole i ʻehā mau pūʻulu: (1) ka hui paʻakai me ka ʻole MIA injection, (2) ka hui MIA me MIA injection, (3) ka hui SDE-treated (200 mg/kg) me ka MIA injection, a me (4 ) ka hui indomethacin- (IM-) i mālama ʻia (2 mg/kg) me MIA injection. Hāʻawi waha ʻia nā ʻiole me SDE a me IM 1 pule ma mua o ka hoʻokomo ʻia ʻana o MIA no 4 mau pule. ʻO ke ʻano o ka SDE a me IM i hoʻohana ʻia i kēia noiʻi ʻana ma luna o nā mea i hana ʻia ma nā haʻawina mua [10,13,14].

2.4.3. Ana o Hindpaw Wehe Kaumaha

Ma hope o ka hoʻokomo ʻana i ka OA, ua hoʻopau ʻia ke koena kumu i ka hiki ke kaumaha o nā hindpaws. Ua hoʻohana ʻia kahi mea hōʻike incapacitance (Linton instrumentation, Norfolk, UK) e loiloi i nā loli i ka hoʻomanawanui ʻana i ke kaumaha. Hoʻokomo pono ʻia nā ʻiole i loko o ke keʻena ana. ʻO ka ikaika kaumaha i hanaʻia e ka lālā hope i hoʻohālikelikeʻia ma kahi o 3 mau manawa. Ua helu ʻia ka lākiō puʻunaue kaumaha e kēia hoohalike: [kaumaha ma ka ʻaoʻao hope ʻākau/(kaumaha ma ka lālā hope ʻākau + kaumaha ma ka lālā hope hema)] × 100 [15].

2.4.4. Nā ana o nā pae Serum Cytokine

Ua centrifuged nā koko koko ma 1,500 g no 10 min ma 4 ° C; a laila ua hōʻiliʻili ʻia ka serum a mālama ʻia ma −70 ° C a hiki i ka hoʻohana ʻana. Nā pae o IL-1β, IL-6, TNF-α, a me PGE2 i loko o ka serum ua ana ʻia me ka hoʻohana ʻana i nā kits ELISA mai R&D Systems (Minneapolis, MN, USA) e like me nā ʻōlelo a ka mea hana.

2.4.5. ʻIkepili RT-PCR ʻOiaʻiʻo

Ua unuhi ʻia ka huina RNA mai ka ʻiʻo hui kuli me ka hoʻohana ʻana i ka TRI reagent® (Sigma-Aldrich, St. Louis, MO, USA), hoʻohuli ʻia i loko o ka cDNA a me ka PCR-amplified me ka hoʻohana ʻana i kahi kit TM One Step RT PCR me SYBR green (Applied Biosystems). , Mokupuni Nui, NY, USA). Ua hana ʻia ka PCR quantitative maoli me ka hoʻohana ʻana i ka Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Grand Island, NY, USA). Hōʻike ʻia nā kaʻina kumu mua a me ke kaʻina noiʻi ma ka Papa1. Ua hoʻonui ʻia nā Aliquots o nā cDNA laʻana a me ka nui like o GAPDH cDNA me ka TaqMan® Universal PCR master mixture i loaʻa ka DNA polymerase e like me nā ʻōlelo a ka mea hana (Applied Biosystems, Foster, CA, USA). ʻO nā kūlana PCR he 2 min ma 50 ° C, 10 min ma 94 ° C, 15 mau minuke ma 95 ° C, a me 1 min ma 60 ° C no 40 mau manawa. Ua hoʻoholo ʻia ka manaʻo o ka gene i hoʻopaʻa ʻia me ka hoʻohana ʻana i ke ʻano Ct (helu pōʻai paepae ma ke ʻano ma waena o ka amplification plot a me ka paepae), e like me nā ʻōlelo a ka mea hana.

Kumukuai FOB:US $0.5 - 9,999 / ʻāpana

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E hoʻouna i ka leka uila iā mākou

Huahana Huahana

ʻO ka Osteoarthritis (OA) ka maʻi musculoskeletal pinepine a me ka maʻi hui degenerative maʻamau i ka poʻe ʻelemakule [1]. ʻO ka OA kahi maʻi i hoʻokumu ʻia e ka ʻeha, ka nalowale o ka hana a me ka hana cartilage, a me ka dysregulation o nā ala proinflammatory a me nā anti-inflammatory.2,3]. Hoʻopilikia nui ia i ka cartilage articular a me ka iwi subchondral o nā hui synovial a me ka hopena i ka hāʻule ʻana o ka hui, e alakaʻi ana i ka ʻeha ma ke kaumaha ʻana me ka hele ʻana a me ke kū ʻana.4].

ʻAʻohe lāʻau lapaʻau no ka OA, no ka mea, he paʻakikī loa ka hoʻihoʻi ʻana i ka cartilage ke luku ʻia.5]. ʻO nā pahuhopu o ka lāʻau lapaʻau e hoʻomaha i ka ʻeha, mālama a hoʻomaikaʻi paha i ka neʻe ʻana o ka hui, hoʻonui i ka ikaika o nā hono, a hoʻemi i nā hopena hōʻino o ka maʻi. ʻO nā lāʻau lapaʻau lāʻau o OA e hoʻemi i ka ʻeha i mea e hoʻonui ai i ka hana hui a me ka maikaʻi o ke ola o ka mea maʻi. ʻOiai ʻo ka luku ʻana i ka cartilage ka mea nui i ka OA, ʻo ka hoʻohaʻahaʻa ʻana o ka collagen ka mea nui e hoʻoholo ai i ka piʻi ʻana o ka OA i hui pū me ka mumū [6,7]. Manaʻo ʻia nā lāʻau lapaʻau me ka hana anti-inflammatory a me ka chondroprotective e hoʻomaha i ka ʻeha a mālama i ka pono matrix i nā maʻi OA.

No laila, ʻoi aku ka maikaʻi o ka hoʻemi ʻana i ka mumū i ka hoʻokele OA. Hōʻike nā haʻawina hou i nā kuleana palekana no nā kumuwaiwai lāʻau i ka holomua o ka OA, ma ke ʻano o ka hoʻohaʻahaʻa ʻana i ka mumū chondrocyte a me ka luku hou ʻana i ka cartilage, ma o ko lākou hiki ke launa pū me nā ʻiʻo i hui pū ʻia, e hopena ai i ka hoʻēmi ʻana i ka ʻeha hui.8].

ʻO ke kumu oʻO Saposhnikovia divaricataUa hoʻohana nui ʻia ʻo Schischkin (Umbelliferae) i ka lāʻau lapaʻau kuʻuna no ka mālama ʻana i ke poʻo, ʻeha, ʻeha, a me ka ʻāʻī ʻana ma Korea a me Kina.9,10]. ʻO nā hopena pharmacological like ʻole oʻO Saposhnikovia divaricata(SD) pū kekahi me nā waiwai anti-inflammatory, analgesic, antipyretic, and antiarthritic properties [9,11]. Ua hōʻike ʻia kahi noiʻi hou loa i loaʻa i ka SD chromone extract nā hopena antirheumatoid arthritis i loko o kahi ʻano ʻiole o ka ʻāʻī ʻana o ka collagen-induced.10]; akā naʻe, kakaikahi nā haʻawina i mālama ʻia e kākoʻo i ka hana anti-inflammatory a me antiarthritis oʻO Saposhnikovia divaricataunuhi (SDE).

No laila, ua noiʻi ka haʻawina i kēia manawa i nā hana anti-inflammatory a me antiosteoarthritis o kahi 70% ethanol extract o SD. ʻO ka mea mua, ua loiloi ʻia ka hopena anti-inflammatory o SDEin vitroi loko o LPS-i hoʻokomo ʻia i ka RAW 264.7 cell. Ma hope aʻe, ua ana ʻia ka hopena antiosteoarthritis o SDE ma ka loiloi ʻana i ka hāʻawi ʻana i ke kaumaha, ka hoʻohaʻahaʻa ʻana o ka cartilage articular, a me nā pane ʻeha i loko o kahi ʻano ʻiole o ka monosodium iodoacetate- (MIA-) i hoʻokomo ʻia i ka OA.