Osteoarthritis (OA) ndi amodzi mwa matenda omwe nthawi zambiri amasokonekera m'mafupa omwe amakhudza anthu azaka zopitilira 65.
1]. Nthawi zambiri, odwala OA amapezeka kuti ali ndi cartilage yowonongeka, synovium yotentha, ndi ma chondrocytes owonongeka, zomwe zimayambitsa ululu ndi kupsinjika maganizo [
2]. Kupweteka kwa nyamakazi kumayamba makamaka chifukwa cha kuwonongeka kwa chichereŵechereŵe m’malo olumikizirana mafupa chifukwa cha kutupa, ndipo chichereŵechereŵenga chikawonongeka kwambiri mafupa amatha kuwombana wina ndi mnzake kumayambitsa kupweteka kosaneneka komanso kuvutikira thupi [
3]. Kuphatikizidwa kwa oyimira pakati otupa omwe ali ndi zizindikiro monga kupweteka, kutupa, ndi kuuma kwa mgwirizano kumalembedwa bwino. Odwala OA, ma cytokines otupa, omwe amachititsa kukokoloka kwa cartilage ndi subchondral bone amapezeka mu synovial fluid.
4]. Madandaulo akulu awiri omwe odwala OA amakhala nawo nthawi zambiri ndi ululu ndi kutupa kwa synovial. Chifukwa chake zolinga zazikulu zamankhwala amakono a OA ndikuchepetsa ululu ndi kutupa. [
5]. Ngakhale mankhwala omwe alipo, kuphatikizapo mankhwala osagwiritsa ntchito steroidal ndi steroidal, atsimikizira kuti amachepetsa ululu ndi kutupa, kugwiritsa ntchito mankhwalawa kwa nthawi yaitali kumakhala ndi zotsatira zoopsa pa thanzi monga mtima, m'mimba, ndi aimpso [
6]. Choncho, mankhwala othandiza kwambiri okhala ndi zotsatira zochepa ayenera kupangidwa pofuna kuchiza nyamakazi ya osteoarthritis.
Zaumoyo zachilengedwe zikuchulukirachulukira kuti ndizotetezeka komanso zopezeka mosavuta [
7]. Mankhwala achikhalidwe aku Korea atsimikizira kuti ali ndi mphamvu zolimbana ndi matenda angapo otupa, kuphatikiza nyamakazi [
8]. Aucklandia lappa DC. amadziwika chifukwa cha mankhwala, monga kupititsa patsogolo kayendedwe ka qi pofuna kuthetsa ululu ndi kuchepetsa m'mimba, ndipo wakhala akugwiritsidwa ntchito ngati mankhwala achilengedwe [
9]. Malipoti am'mbuyomu akuwonetsa kuti A. lappa ali ndi anti-inflammatory [
10,
11], ochepetsa ululu [
12], anticancer [
13] ndi gastroprotective [
14] zotsatira. Zochita zosiyanasiyana zamoyo za A. lappa zimayambitsidwa ndi mankhwala ake akuluakulu: costunolide, dehydrocostus lactone, dihydrocostunolide, costuslactone, α-costol, saussurea lactone ndi costuslactone [
15]. Kafukufuku wam'mbuyomu amati costunolide inawonetsa anti-inflammatory properties mu lipopolysaccharide (LPS), yomwe inachititsa macrophages kupyolera mu kayendetsedwe ka NF-kB ndi njira ya mapuloteni a kutentha.
16,
17]. Komabe, palibe kafukufuku yemwe wafufuza ntchito zomwe A. lappa angachite pochiza OA. Kafukufuku wapano wafufuza za chithandizo cha A. lappa motsutsana ndi OA pogwiritsa ntchito (monosodium-iodoacetate) MIA ndi mitundu ya makoswe a acetic acid.
Monosodium-iodoacetate (MIA) imagwiritsidwa ntchito kwambiri popanga zowawa zambiri komanso mawonekedwe a pathophysiological a OA mu nyama.
18,
19,
20]. Pamene jekeseni m'magulu a mawondo, MIA imasokoneza kagayidwe kake ka chondrocyte ndipo imayambitsa kutupa ndi zizindikiro zotupa, monga cartilage ndi subchondral bone erosion, zizindikiro zazikulu za OA [
18]. Kuyankha kwamphamvu komwe kumapangidwa ndi acetic acid kumawonedwa mofala ngati kuyerekezera kwa ululu wam'mphepete mwa nyama komwe ululu wotupa ukhoza kuyeza mochulukira [
19]. Mzere wa cell wa mbewa wa macrophage, RAW264.7, umagwiritsidwa ntchito kwambiri pophunzira mayankho amtundu wa kutupa. Pambuyo poyambitsa ndi LPS, macrophages a RAW264 amachititsa njira zotupa ndipo amatulutsa oyimira angapo otupa, monga TNF-α, COX-2, IL-1β, iNOS, ndi IL-6 [
20]. Kafukufukuyu adawunikira zotsatira za anti-nociceptive ndi anti-inflammatory za A. lappa motsutsana ndi OA mu MIA chitsanzo cha nyama, chitsanzo cha acetic acid-induced animal model, ndi LPS-activated RAW264.7 maselo.
2. Zida ndi Njira
2.1. Zomera Zomera
Muzu wouma wa A. lappa DC. zomwe zidagwiritsidwa ntchito pakuyesa zidagulidwa ku Epulip Pharmaceutical Co., Ltd., (Seoul, Korea). Zinadziwika ndi Prof. Donghun Lee, Dept. of Herbal pharmacology, Col. of Korean Medicine, Gachon University, ndipo nambala ya voucher inayikidwa ngati 18060301.
2.2. HPLC Analysis of A. lappa Extract
A. lappa adatengedwa pogwiritsa ntchito chipangizo cha reflux (madzi osungunuka, 3 h pa 100 °C). Njira yochotsera idasefedwa ndikufupikitsidwa pogwiritsa ntchito evaporator yotsika mphamvu. Chotsitsa cha A. lappa chinali ndi zokolola za 44.69% pambuyo poumitsa-kuzizira pansi pa −80 °C. Kusanthula kwa Chromatographic kwa A. lappa kunachitika ndi HPLC yolumikizidwa pogwiritsa ntchito 1260 InfinityⅡ HPLC-system (Agilent, Pal Alto, CA, USA). Pakupatukana kwa chromatic, EclipseXDB C18 column (4.6 × 250 mm, 5 µm, Agilent) idagwiritsidwa ntchito pa 35 °C. Okwana 100 mg wa chitsanzo anali kuchepetsedwa mu 10 mL wa 50% methanol ndi sonicated 10 min. Zitsanzo zidasefedwa ndi sefa ya syringe (Waters Corp., Milford, MA, USA) ya 0.45 μm. Mapangidwe a gawo la mafoni anali 0.1% phosphoric acid (A) ndi acetonitrile (B) ndipo gawoli linatulutsidwa motere: 0-60 min, 0%; 60-65 min, 100%; 65-67 min, 100%; 67-72 min, 0% zosungunulira B ndi kuthamanga kwa 1.0 mL / min. Madzi otayira amawonedwa pa 210 nm pogwiritsa ntchito jekeseni wa 10 μL. Kusanthula kunachitika katatu.
2.3. Nyumba ndi Kasamalidwe ka Zinyama
Makoswe aamuna a Sprague-Dawley (SD) azaka za masabata a 5 ndi mbewa zamphongo za ICR zazaka za 6 zidagulidwa kuchokera ku Samtako Bio Korea (Gyeonggi-do, Korea). Zinyama zinkasungidwa m'chipinda pogwiritsa ntchito kutentha kosasintha (22 ± 2 ° C) ndi chinyezi (55 ± 10%) ndi kuwala / mdima wa 12/12 h. Nyamazo zinkadziwika bwino ndi vutoli kwa nthawi yoposa sabata imodzi isanayambe kuyesa. Nyama zinali ndi ad libitum kupereka chakudya ndi madzi. Malamulo amakono osamalira ndi kusamalira nyama ku Yunivesite ya Gachon (GIACUC-R2019003) adatsatiridwa mosamalitsa pamachitidwe onse oyesera nyama. Phunzirolo linapangidwa kuti likhale loyesa-khungu komanso lofanana. Tinatsatira njira ya euthanasia molingana ndi malangizo a Komiti Yoyeserera Yanyama.
2.4. Jekeseni wa MIA ndi Chithandizo
Makoswe adagawidwa mwachisawawa m'magulu anayi, omwe ndi sham, control, indomethacin, ndi A. lappa. Pokhala ndi anesthetized ndi 2% isofluorane O2 osakaniza, makoswe anabayidwa pogwiritsa ntchito 50 μL ya MIA (40 mg / m; Sigma-Aldrich, St. Louis, MO, USA) intra-articularly m'magulu a mawondo kuti atsogolere ku OA yoyesera. Mankhwalawa adachitidwa monga pansipa: magulu olamulira ndi achinyengo adasungidwa kokha ndi zakudya zoyambira za AIN-93G. Pokhapokha, gulu la indomethacin linapatsidwa indomethacin (3 mg/kg) yophatikizidwa muzakudya za AIN-93G ndipo gulu la A. lappa 300 mg/kg linapatsidwa chakudya cha AIN-93G chowonjezera ndi A. lappa (300 mg/kg). Mankhwalawa adapitilizidwa kwa masiku 24 kuyambira tsiku la kulowetsedwa kwa OA pamlingo wa 15-17 g pa 190-210 g kulemera kwa thupi tsiku lililonse.
2.5. Kuyeza Kulemera kwake
Pambuyo pa kulowetsedwa kwa OA, kuyeza kulemera kwa miyendo yakumbuyo kwa makoswe kunachitidwa ndi incapacitance-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) monga momwe anakonzera. Kugawa kulemera pamiyendo yakumbuyo kunawerengedwa: kunyamula kulemera (%)
