I-Osteoarthritis (OA) ingesinye sezifo ezihlala isikhathi eside eziwohlokayo zamathambo ezithinta abantu abadala abangaphezu kweminyaka engama-65 [
1]. Ngokuvamile, iziguli ze-OA kutholakala ukuthi zine-cartilage eyonakele, i-synovium evuthayo, nama-chondrocyte agugulekile, okubangela ubuhlungu nokucindezeleka ngokomzimba [
2]. Ubuhlungu be-arthritis bubangelwa kakhulu ukuwohloka koqwanga emalungeni ngenxa yokuvuvukala, futhi lapho uqwanga lonakele kakhulu amathambo angashayisana abangele ubuhlungu obungabekezeleleki nobunzima bomzimba [
3]. Ukubandakanyeka kwabalamuli abavuvukalayo abanezimpawu ezinjengobuhlungu, ukuvuvukala, nokuqina kwelunga kubhalwe kahle. Ezigulini ze-OA, ama-cytokines avuvukalayo, abangela ukuguguleka kwe-cartilage ne-subchondral bone atholakala ku-synovial fluid [
4]. Izikhalazo ezimbili ezinkulu iziguli ze-OA ezivame ukuba nazo ubuhlungu nokuvuvukala kwe-synovial. Ngakho-ke imigomo eyinhloko yezindlela zokwelapha ze-OA zamanje ukunciphisa ubuhlungu nokuvuvukala. [
5]. Nakuba izindlela zokwelapha ze-OA ezitholakalayo, okuhlanganisa nezidakamizwa ezingezona eze-steroidal kanye ne-steroidal, ziye zafakazela ukuthi ziyasebenza ekudambiseni ubuhlungu nokuvuvukala, ukusetshenziswa kwesikhathi eside kwale mithi kunemiphumela emibi kakhulu yezempilo njengokungasebenzi kahle kwenhliziyo, i-gastro-intestinal, kanye nezinso [
6]. Ngakho-ke, umuthi osebenza ngempumelelo onemiphumela engemihle embalwa kufanele uthuthukiswe ukwelapha i-osteoarthritis.
Imikhiqizo yezempilo yemvelo iya ngokuya iduma ngokuphepha nokutholakala kalula [
7]. Imithi yendabuko yaseKorea ifakazele ukuthi iyasebenza ekulweni nezifo eziningana ezivuvukalayo, kuhlanganise nesifo samathambo [
8]. I-Aucklandia lappa DC. yaziwa ngezici zayo zokwelapha, njengokuthuthukisa ukujikeleza kwe-qi ukuze kudambise izinhlungu nokupholisa isisu, futhi isetshenziswe ngokwesiko njenge-analgesic yemvelo [
9]. Imibiko yangaphambilini iphakamisa ukuthi i-A. lappa ine-anti-inflammatory [
10,
11], i-analgesic [
12], umdlavuza [
13], kanye ne-gastroprotective [
14] imiphumela. Imisebenzi ehlukahlukene yezinto eziphilayo ye-A. lappa ibangelwa izinhlanganisela zayo ezinkulu ezisebenzayo: i-costunolide, i-dehydrocostus lactone, i-dihydrocostunolide, i-costuslactone, i-α-costol, i-saussurea lactone ne-costuslactone [
15]. Ucwaningo lwangaphambilini luthi i-costunolide ibonise izakhiwo zokulwa nokuvuvukala ku-lipopolysaccharide (LPS), eyenza ama-macrophages ngokusebenzisa ukulawulwa kwe-NF-kB kanye nomzila weprotheyini wokushisa ukushisa [
16,
17]. Nokho, alukho ucwaningo oluphenye imisebenzi engaba khona ye-A. lappa yokwelashwa kwe-OA. Ucwaningo lwamanje luphenye imiphumela yokwelapha ye-A. lappa ngokumelene ne-OA kusetshenziswa (i-monosodium-iodoacetate) i-MIA namamodeli wamagundane enziwe nge-acetic acid.
I-Monosodium-iodoacetate (MIA) isetshenziswa ngokudumile ukukhiqiza okuningi kokuziphatha kobuhlungu kanye nezici ze-pathophysiological ze-OA ezilwaneni [
18,
19,
20]. Lapho ijova emalungeni amadolo, i-MIA iphazamisa i-chondrocyte metabolism futhi ibangele ukuvuvukala nezimpawu zokuvuvukala, njengokuguguleka kwe-cartilage kanye ne-subchondral bone, izimpawu eziyinhloko ze-OA [
18]. Ukuphendula okubhaliwe okwenziwe nge-acetic acid kubhekwa kabanzi njengokulingisa ubuhlungu be-peripheral ezilwaneni lapho ubuhlungu obuvuthayo bungalinganiswa ngobuningi [
19]. I-mouse macrophage cell line, i-RAW264.7, isetshenziswa kakhulu ukutadisha izimpendulo zamaselula ekuvuvukeni. Lapho kusebenze nge-LPS, ama-macrophage e-RAW264 avula izindlela zokuvuvukala futhi afihle abaxhumanisi abaningana abavuvukalayo, njenge-TNF-α, COX-2, IL-1β, iNOS, ne-IL-6 [
20]. Lolu cwaningo luhlole imiphumela ephikisana ne-nociceptive ne-anti-inflammatory ye-A. lappa ngokumelene ne-OA kumodeli yezilwane ye-MIA, imodeli yezilwane eyenziwe nge-acetic acid, kanye namaseli e-RAW264.7 acushwe yi-LPS.
2. Izinto kanye Nezindlela
2.1. Impahla Yezitshalo
Impande eyomile ye-A. lappa DC. okusetshenziswe ekuhloleni kuthengwe kwa-Epulip Pharmaceutical Co., Ltd., (Seoul, Korea). Yahlonzwa nguProf. Donghun Lee, uMnyango we-Herbal pharmacology, Col. of Korean Medicine, Gachon University, futhi inombolo yesampula yevawusha yafakwa ngokuthi 18060301.
2.2. Ukuhlaziywa kwe-HPLC kwe-A. lappa Extract
I-A. lappa yakhishwa kusetshenziswa i-reflux apparatus (amanzi acwecwe, 3 h ku-100 °C). Isixazululo esikhishiwe sahlungwa futhi safinyezwa kusetshenziswa i-evaporator enengcindezi ephansi. I-A. lappa extract ibe nesivuno esingu-44.69% ngemva kokumiswa yiqhwa ngaphansi kokungu-−80 °C. Ukuhlaziywa kweChromatographic kwe-A. lappa kwenziwe nge-HPLC exhunywe kusetshenziswa i-1260 InfinityⅡ HPLC-system (Agilent, Pal Alto, CA, USA). Ngokuhlukaniswa kwe-chromatic, ikholomu ye-EclipseXDB C18 (4.6 × 250 mm, 5 µm, Agilent) isetshenziswe ku-35 °C. Isamba esingu-100 mg wesampula sihlanjululwe ku-10 mL we-methanol engu-50% futhi yafakwa i-sonication imizuzu engu-10. Amasampuli ahlungwa ngesihlungi sesirinji (Waters Corp., Milford, MA, USA) esingu-0.45 μm. Ukwakheka kwesigaba seselula bekuyi-0.1% phosphoric acid (A) ne-acetonitrile (B) futhi ikholomu yacaciswa kanje: 0–60 min, 0%; 60-65 amaminithi, 100%; 65–67 min, 100%; 67–72 min, 0% i-solvent B enezinga lokugeleza lika-1.0 mL/min. Ukungcola kubonwe ku-210 nm kusetshenziswa umthamo womjovo ongu-10 μL. Ukuhlaziya kwenziwe ngokuphindwe kathathu.
2.3. Izindlu Zezilwane Nokuphathwa
Amagundane angama-Sprague–Dawley (SD) anamaviki angu-5 kanye namagundane angama-ICR anamaviki angu-6 athengwe kwaSamtako Bio Korea (Gyeonggi-do, Korea). Izilwane zazigcinwa ekamelweni kusetshenziswa izinga lokushisa elingaguquki (22 ± 2 °C) kanye nomswakama (55 ± 10%) kanye nomjikelezo wokukhanya/omnyama we-12/12 h. Izilwane zazijwayelene nalesi simo isikhathi esingaphezu kwesonto ngaphambi kokuba kuqale ukuhlolwa. Izilwane zazine-ad libitum supply yokuphakelayo namanzi. Imithetho yokuziphatha yamanje yokunakekelwa nokuphathwa kwezilwane e-Gachon University (GIACUC-R2019003) ilandelwe ngokuqinile kuzo zonke izinqubo zokuhlola izilwane. Ucwaningo lwenzelwe uphenyo oluphuphuthekiswe umphenyi noluhambisanayo. Silandele indlela yokubulala ngokweziqondiso zeKomidi Lokuziphatha Lokuhlola Izilwane.
2.4. I-MIA Injection Nokwelashwa
Amagundane ahlukaniswa ngokungahleliwe abe amaqembu angu-4, okungukuthi sham, control, indomethacin, kanye ne-A. lappa. Njengoba ekhishwa izinzwa ngengxube ye-2% ye-isofluorane O2, amagundane ajovwa kusetshenziswa i-50 μL ye-MIA (40 mg/m; Sigma-Aldrich, St. Louis, MO, USA) intra-articularly emalungeni amadolo ukuze aholele ku-OA yokuhlola. Ukwelashwa kwenziwa ngezansi: amaqembu okulawula kanye nama-sham agcinwe kuphela nge-AIN-93G yokudla okuyisisekelo. Kuphela, iqembu le-indomethacin linikezwe i-indomethacin (3 mg/kg) ehlanganiswe nokudla kwe-AIN-93G kanye neqembu le-A. lappa 300 mg/kg labelwa ekudleni kwe-AIN-93G okungezwe nge-A. lappa (300 mg/kg). Ukwelashwa kwaqhutshekwa izinsuku ezingu-24 kusukela ngosuku lokungeniswa kwe-OA ngenani lika-15-17 g nge-190-210 g yesisindo somzimba nsuku zonke.
2.5. Isisindo Sokuthwala
Ngemva kokufakwa kwe-OA, ukukalwa kwamandla okuthwala isisindo semilenze yangemuva yamagundane kwenziwa nge-incapacitance-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) njengoba kuhleliwe. Ukusatshalaliswa kwesisindo ezithweni zangemuva kubaliwe: umthamo wokuthwala isisindo (%)
