ʻAila Saposhnikovia divaricata maʻemaʻe no ka hana ʻana i ke kukui a me ke kopa, ka mea hoʻolaha nui o ka ʻaila koʻikoʻi hou no nā mea hoʻolaha puhi ʻōheu

ʻAila Saposhnikovia divaricata maʻemaʻe no ka hana ʻana i ke kukui a me ke kopa, ka mea hoʻolaha nui, ka ʻaila koʻikoʻi hou no nā mea hoʻolaha puhi ʻōheu. Kiʻi i hōʻike ʻia

wehewehe pōkole:

2.1. Hoʻomākaukau ʻana o SDE

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

2.2. Ka Nānā ʻana i ka Chromatography Wai Hana Kiʻekiʻe (HPLC)

Ua hana ʻia ka loiloi chromatographic me kahi ʻōnaehana HPLC (Waters Co., Milford, MA, USA) a me kahi mea ʻike photodiode array. No ka loiloi HPLC o SDE, ʻo ka prim-O-ua kūʻai ʻia ke kūlana glucosylcimifugin mai ka Korea Promotion Institute for Traditional Medicine Industry (Gyeongsan, Korea), akekona-O-glucosylhamaudol a me 4′-O-β-D-glucosyl-5-OUa hoʻokaʻawale ʻia ka -methylvisamminol i loko o kā mākou keʻena hoʻokolohua a ua ʻike ʻia e nā loiloi spectral, ʻo ia hoʻi e NMR a me MS.

Ua hoʻoheheʻe ʻia nā laʻana SDE (0.1 mg) i loko o 70% ethanol (10 mL). Ua hana ʻia ka hoʻokaʻawale chromatographic me kahi kolamu XSelect HSS T3 C18 (4.6 × 250 mm, 5μm, Waters Co., Milford, MA, USA). ʻO ka pae lawe lima he acetonitrile (A) a me 0.1% acetic acid i loko o ka wai (B) ma ka nui o ke kahe ʻana 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 scan ʻia ka nalu ʻike ma 210–400 nm a hoʻopaʻa ʻia ma 254 nm. ʻO ka nui o ka injection he 10.0μL. Ua hoʻomākaukau ʻia nā hopena maʻamau no ka hoʻoholo ʻana o ʻekolu chromones ma kahi hoʻohui hope loa o 7.781 mg/mL (prim-O-glucosylimifugin), 31.125 mg/mL (4′-O-β-D-glucosyl-5-O-methylvisamminol), a me 31.125 mg/mL (kekona-O-glucosylhamaudol) i loko o ka methanol a mālama ʻia ma 4°C.

2.3. Ka Loiloi ʻana o ka Hana Anti-InflammatoryI loko o ka Vitro

2.3.1. Ka Moʻomeheu Kelepona a me ka Lapaʻau ʻana i nā Laʻana

Ua loaʻa nā pūnaewele RAW 264.7 mai ka American Type Culture Collection (ATCC, Manassas, VA, USA) a ua ulu ʻia i loko o ka medium DMEM e loaʻa ana he 1% antibiotics a me 5.5% FBS. Ua hoʻoulu ʻia nā pūnaewele i loko o kahi lewa haʻahaʻa o 5% CO2 ma 37°C. No ka hoʻoulu ʻana i nā pūnaewele, ua pani ʻia ka medium me ka medium DMEM hou, a me ka lipopolysaccharide (LPS, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) ma 1.μua hoʻohui ʻia ʻo g/mL i ke alo a i ʻole ka ʻole o SDE (200 a i ʻole 400μg/mL) no 24 hola hou aku.

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

Ua mālama ʻia nā pūnaewele me SDE a hoʻoulu ʻia me LPS no 24 mau hola. Ua kālailai ʻia ka hana ʻana o NO ma ke ana ʻana i ka nitrite me ka hoʻohana ʻana i ka reagent Griess e like me kahi noiʻi mua [12Hoʻokuʻu ʻia ʻana o nā cytokines mumū PGE2, TNF-α, a ua hoʻoholo ʻia ʻo IL-6 me ka hoʻohana ʻana i kahi pahu ELISA (nā ʻōnaehana R&D) e like me nā kuhikuhi a ka mea hana. Ua hoʻoholo ʻia nā hopena o SDE ma ka hana ʻana o NO a me ka 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 ʻana o ka Hana AntiosteoarthritisI loko o Vivo

2.4.1. Nā holoholona

Ua kūʻai ʻia nā ʻiole Sprague-Dawley kāne (7 pule) mai Samtako Inc. (Osan, Korea) a ua hoʻonoho ʻia ma lalo o nā kūlana i kāohi ʻia me kahi pōʻaiapuni mālamalama/pouli 12-h ma°C a me% haumākū. Ua hāʻawi ʻia nā ʻiole i ka ʻai a me ka wai ma ka hale hanahoʻolaha kūʻokoʻaUa hana ʻia nā kaʻina hana hoʻokolohua a pau e like me nā alakaʻi o ka National Institutes of Health (NIH) a ua ʻāpono ʻia e ke Kōmike Mālama a me ka Hoʻohana Holoholona o ke kulanui ʻo Daejeon (Daejeon, repubalika o Korea).

2.4.2. Hoʻokomo ʻia ʻana o OA me MIA i nā ʻiole

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

2.4.3. Nā Ana o ka Hoʻolaha Kaumaha o ka Hindpaw

Ma hope o ka hoʻokomo ʻia ʻana o OA, ua hoʻopilikia ʻia ke kaulike mua i ka hiki ke hāpai i ke kaumaha o nā wāwae hope. Ua hoʻohana ʻia kahi mea hoʻāʻo incapacitance (Linton instrumentation, Norfolk, UK) e loiloi i nā loli i ke ahonui hāpai kaumaha. Ua hoʻokomo pono ʻia nā ʻiole i loko o ke keʻena ana. Ua hoʻohālikelike ʻia ka ikaika hāpai kaumaha i hoʻokau ʻia e ka lālā hope ma luna o kahi manawa 3 s. Ua helu ʻia ka lakio hoʻolaha kaumaha e ka hoʻohālikelike penei: [kaumaha ma ka lālā hope ʻākau/(kaumaha ma ka lālā hope ʻākau + kaumaha ma ka lālā hope hema)] × 100 [15].

2.4.4. Nā Ana o nā Pae Cytokine Serum

Ua centrifuged ʻia nā hāpana 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. ʻO nā pae o IL-1β, IL-6, TNF-α, a ua ana ʻia ʻo PGE2 i loko o ke serum me ka hoʻohana ʻana i nā kime ELISA mai R&D Systems (Minneapolis, MN, USA) e like me nā kuhikuhi a ka mea hana.

2.4.5. Ka Nānā ʻana i ka RT-PCR Manawa Maoli

Ua unuhi ʻia ka RNA holoʻokoʻa mai nā ʻiʻo hui kuli me ka hoʻohana ʻana i ka TRI reagent® (Sigma-Aldrich, St. Louis, MO, USA), i hoʻololi ʻia i loko o cDNA a hoʻonui ʻia i ka PCR me ka hoʻohana ʻana i kahi pahu TM One Step RT PCR me SYBR ʻōmaʻomaʻo (Applied Biosystems, Grand Island, NY, USA). Ua hana ʻia ka PCR quantitative manawa maoli me ka hoʻohana ʻana i ka ʻōnaehana Applied Biosystems 7500 Real-Time PCR (Applied Biosystems, Grand Island, NY, USA). Hōʻike ʻia nā moʻo primer a me ka moʻo probe ma ka Papa.1. Ua hoʻonui ʻia nā ʻāpana o nā cDNA laʻana a me ka nui like o ka GAPDH cDNA me ka hui haku TaqMan® Universal PCR e loaʻa ana ka DNA polymerase e like me nā kuhikuhi 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 s ma 95°C, a me 1 min ma 60°C no 40 mau pōʻaiapuni. Ua hoʻoholo ʻia ka nui o ka gene target me ka hoʻohana ʻana i ke ʻano hoʻohālikelike Ct (helu pōʻaiapuni paepae ma ke kiko keʻa ma waena o ka hoʻolālā amplification a me ka paepae), e like me nā kuhikuhi a ka mea hana.

Kumukūʻai FOB:US $0.5 - 9,999 / ʻĀpana

Ka nui o ke kauoha liʻiliʻi loa:100 ʻāpana/ʻāpana

Ka hiki ke hoʻolako:10000 ʻāpana/ʻāpana i kēlā me kēia mahina

E leka uila mai iā mākou

Nā kikoʻī huahana

Nā Lepili Huahana

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

ʻAʻohe lāʻau lapaʻau no ka OA, no ka mea, he mea paʻakikī loa ke hoʻihoʻi i ke cartilage ke pau i ka luku ʻia.5ʻO nā pahuhopu o ka mālama ʻana, ʻo ia ka hōʻoluʻolu ʻana i ka ʻeha, mālama a hoʻomaikaʻi paha i ka neʻe ʻana o nā hono, hoʻonui i ka ikaika o nā hono, a hoʻēmi i nā hopena hoʻopilikia o ka maʻi. ʻO ka pahuhopu o nā lāʻau lapaʻau o OA e hōʻemi i ka ʻeha i mea e hoʻonui ai i ka hana hui o ka mea maʻi a me ke ʻano o ke ola. ʻOiai ʻo ka luku ʻia ʻana o ke cartilage ka hanana nui ma OA, ʻo ka hōʻino ʻia o ka collagen ka hanana nui e hoʻoholo ai i ka holomua ʻole o OA e pili ana i ka mumū [6,7Manaʻo ʻia nā lāʻau lapaʻau me ka hana anti-inflammatory a me chondroprotective e hōʻoluʻolu i ka ʻeha a mālama i ka pono o ka matrix i nā poʻe maʻi OA.

No laila, ʻo ka hoʻemi ʻana i ka mumū e lilo paha i mea pono i ka hoʻokele OA. Hōʻike nā haʻawina hou i nā kuleana palekana no nā kumuwaiwai lāʻau lapaʻau i ka holomua o OA, ma ke ʻano o ka hōʻemi ʻana i ka mumū chondrocyte a me ka luku hou ʻana o ka cartilage, ma o ko lākou hiki ke launa pū me nā ʻiʻo e pili ana i nā hono, e hopena ana i ka hōʻemi ʻana i ka ʻeha hui [8].

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

No laila, ua noiʻi ke aʻo ʻana 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 SDEi loko o ka lumi hoʻomoʻai loko o nā pūnaewele RAW 264.7 i hoʻokomo ʻia e LPS. ʻO ka mea aʻe, ua ana ʻia ka hopena antiosteoarthritis o SDE ma ka loiloi ʻana i ka hoʻolaha ʻana o ke kaumaha, ka hōʻino ʻana o ke cartilage articular, a me nā pane mumū i loko o kahi kumu hoʻohālike iole o ka monosodium iodoacetate- (MIA-) i hoʻokomo ʻia ʻo OA.