I-osteoarthritis (OA) sesinye sezifo zexesha elide ezingapheliyo ezichaphazela amathambo abantu abadala abangaphezu kweminyaka engama-65.
1]. Ngokuqhelekileyo, izigulane ze-OA zifunyaniswa zine-cartilage eyonakeleyo, i-synovium evuthayo, kunye ne-chondrocytes ekhukhulayo, ebangela intlungu kunye nokukhathazeka ngokomzimba [
2]. Intlungu yeArthritic ibangelwa kakhulu kukuwohloka kwe-cartilage kumalungu ngenxa yokudumba, kwaye xa i-cartilage yonakaliswe kakhulu amathambo anokubetha omnye komnye kubangele iintlungu ezinganyamezelekiyo kunye nobunzima bomzimba [
3]. Ukubandakanyeka kwabalamli abavuthayo abaneempawu ezifana nentlungu, ukuvuvukala, kunye nokuqina komgudu kubhalwe kakuhle. Kwizigulane ze-OA, i-cytokines evuthayo, ebangela ukuguguleka kwe-cartilage kunye ne-subchondral bone ifumaneka kwi-synovial fluid [
4]. Izikhalazo ezibini eziphambili ezithi izigulane ze-OA zibe nazo ngokubanzi ziintlungu kunye nokudumba kwe-synovial. Ke ngoko iinjongo eziphambili zolu nyango lwangoku lwe-OA kukuthoba iintlungu kunye nokudumba. [
5]. Nangona unyango olukhoyo lwe-OA, kubandakanywa amachiza angeyo-steroidal kunye ne-steroidal, abonakalise ukusebenza kakuhle ekuthomalaliseni iintlungu kunye nokudumba, ukusetyenziswa kwexesha elide kwala machiza kuneziphumo ezibi zempilo ezinje nge-cardiovascular, gastro-intestinal, kunye nokungasebenzi kakuhle kwezintso [
6]. Ke ngoko, iyeza elisebenzayo elineziphumo ebezingalindelekanga ezimbalwa kufuneka lenziwe kunyango lwe-osteoarthritis.
Iimveliso zempilo zendalo ziya zithandwa kakhulu ukuze zikhuseleke kwaye zifumaneke lula [
7]. Amayeza emveli aseKorea abonakalise ukusebenza ngempumelelo ngokuchasene nezifo ezininzi ezivuthayo, kubandakanya isifo samathambo [
8]. Aucklandia lappa DC. iyaziwa ngeempawu zayo zonyango, ezifana nokuphucula ukujikeleza kwe-qi ukukhulula intlungu kunye nokuthomalalisa isisu, kwaye isetyenziswe ngokwesiko njenge-analgesic yendalo [
9]. Iingxelo zangaphambili zibonisa ukuba i-A. lappa ine-anti-inflammatory [
10,
11], i-analgesic [
12], umhlaza [
13], kunye nokukhusela isisu [
14] iziphumo. Imisebenzi eyahlukeneyo yebhayoloji ye-A. lappa ibangelwa zizixhobo zayo eziphambili ezisebenzayo: i-costunolide, i-dehydrocostus lactone, i-dihydrocostunolide, i-costuslactone, i-α-costol, i-saussurea lactone kunye ne-costuslactone [
15]. Uphononongo lwangaphambili lubanga ukuba i-costunolide ibonise iipropati ezichasayo kwi-lipopolysaccharide (LPS), eyenza i-macrophages ngolawulo lwe-NF-kB kunye neprotheyini yokutshatyalaliswa kobushushu [
16,
17]. Nangona kunjalo, akukho phando luphande imisebenzi enokwenzeka ye-A. lappa yonyango lwe-OA. Uphando lwangoku luphande iziphumo zonyango lwe-A. lappa ngokuchasene ne-OA kusetyenziswa (i-monosodium-iodoacetate) i-MIA kunye neemodeli ze-acetic-induced rodent.
I-Monosodium-iodoacetate (MIA) isetyenziswa ngokudumileyo ukuvelisa uninzi lweentlungu zokuziphatha kunye neempawu ze-pathophysiological ze-OA kwizilwanyana [.
18,
19,
20]. Xa ijojowe kumalungu amadolo, i-MIA iphazamisa i-chondrocyte metabolism kwaye ibangele ukuvuvukala kunye neempawu ezivuthayo, ezifana ne-cartilage kunye ne-subchondral bone erosion, iimpawu zekhadinali ze-OA [
18]. Ukuphendula okubhaliweyo okubangelwa i-acetic acid kuthathwa ngokubanzi njengokulinganisa intlungu ye-peripheral kwizilwanyana apho intlungu evuthayo inokulinganiswa ngokobungakanani [
19]. I-mouse macrophage cell line, i-RAW264.7, isetyenziselwa ukufunda iimpendulo zeselula ekudumbeni. Emva kokusebenza kunye ne-LPS, i-RAW264 i-macrophages ivula iindlela ezivuthayo kwaye ikhuphe ama-intermediaries amaninzi avuthayo, njenge-TNF-α, i-COX-2, i-IL-1β, i-iNOS, kunye ne-IL-6 [
20]. Olu pho nonongo luye lwavavanya i-anti-nociceptive kunye ne-anti-inflammatory effects ye-A. lappa ngokuchasene ne-OA kwimodeli yezilwanyana ze-MIA, imodeli yezilwanyana ze-acetic acid, kunye ne-LPS-activated RAW264.7 cells.
2. Izinto kunye neendlela
2.1. Izixhobo zezityalo
Ingcambu eyomileyo ye-A. lappa DC. isetyenziswe kuvavanyo yathengwa kwi-Epulip Pharmaceutical Co., Ltd., (Seoul, Korea). Yachongwa nguProf. Donghun Lee, iSebe le-Herbal pharmacology, Col. of Korean Medicine, iYunivesithi yaseGachon, kunye nenombolo yomzekelo wevawutsha yafakwa njenge-18060301.
2.2. Uhlalutyo lwe-HPLC lwe-A. lappa Extract
I-lappa ikhutshwe kusetyenziswa i-reflux apparatus (amanzi adibeneyo, i-3 h kwi-100 ° C). Isisombululo esikhutshwayo sahluzwa kwaye sancitshiswa kusetyenziswa i-evaporator yoxinzelelo oluphantsi. Isicatshulwa se-A. lappa sibe nesivuno se-44.69% emva kokumiswa komkhenkce ngaphantsi -80 °C. Uhlalutyo lweChromatographic ye-A. lappa lwenziwe nge-HPLC eqhagamshelwe kusetyenziswa i-1260 InfinityⅡ HPLC-system (Agilent, Pal Alto, CA, USA). Ukwahlula ikhromatic, ikholamu ye-EclipseXDB C18 (4.6 × 250 mm, 5 µm, Agilent) isetyenziswe kuma-35 °C. Ingqikithi ye-100 mg ye-specimen yahlanjululwa kwi-10 mL ye-50% ye-methanol kunye ne-sonicated ye-10 min. Iisampulu zahluzwa ngesihluzo sesirinji (Waters Corp., Milford, MA, USA) ye-0.45 μm. Ukubunjwa kwesigaba seselula kwakuyi-0.1% ye-phosphoric acid (A) kunye ne-acetonitrile (B) kwaye ikholamu yahlanjululwa ngolu hlobo lulandelayo: 0-60 min, 0%; 60–65 min, 100%; 65–67 min, 100%; I-67-72 min, i-0% ye-solvent B kunye nesantya sokuhamba kwe-1.0 mL / min. Amanzi amdaka ajongwe kwi-210 nm kusetyenziswa umthamo we-naliti ye-10 μL. Uhlalutyo lwenziwa kathathu.
2.3. Izindlu zezilwanyana kunye noLawulo
Iigundane ze-Sprague-Dawley (SD) ezineminyaka eneeveki ze-5 kunye neempuku ze-ICR eziyindoda ezineminyaka eyi-6 iiveki zathengwa kwi-Samtako Bio Korea (Gyeonggi-do, Korea). Izilwanyana zazigcinwa kwigumbi kusetyenziswa ubushushu obungaguqukiyo (22 ± 2 °C) kunye nokufuma (55 ± 10%) kunye nomjikelo wokukhanya/omnyama we-12/12 h. Izilwanyana zaziqhelene nale meko ixesha elingaphezu kweveki ngaphambi kokuba kuqaliswe uvavanyo. Izilwanyana zazinobonelelo lwe-ad libitum yokutya kunye namanzi. Imithetho yokuziphatha yangoku yokunakekelwa kwezilwanyana kunye nokuphathwa kweYunivesithi yaseGachon (GIACUC-R2019003) ilandelwe ngokungqongqo kuzo zonke iinkqubo zovavanyo lwezilwanyana. Uphononongo lwenzelwe umphandi-mfameke kunye nolingo oluhambelanayo. Salandela indlela ye-euthanasia ngokwemigaqo ye-Animal Experimental Ethics Committee.
2.4. Isitofu se-MIA kunye noNyango
Iimpuku zahlulwa ngokungenamkhethe zibe ngamaqela ama-4, angala, i-sham, ulawulo, i-indomethacin, kunye ne-A. lappa. Ukuba i-anesthetized kunye ne-2% ye-isofluorane umxube we-O2, iigundane zajojowe ngokusebenzisa i-50 μL ye-MIA (40 mg / m; Sigma-Aldrich, St. Louis, MO, USA) intra-articularly kwi-knee joints ukukhokelela kwi-OA yokuhlola. Unyango luqhutywe ngolu hlobo lungezantsi: ulawulo kunye namaqela e-sham agcinwe kuphela nge-AIN-93G yokutya okusisiseko. Kuphela, iqela le-indomethacin libonelelwe nge-indomethacin (3 mg / kg) efakwe kwi-AIN-93G yokutya kunye ne-A. lappa 300 mg / kg iqela labelwa kwi-AIN-93G yokutya eyongezwe nge-A. lappa (300 mg / kg). Unyango lwaqhutyekwa kwiintsuku ezingama-24 ukususela kwimini ye-OA yokufakwa kwizinga le-15-17 g nge-190-210 g ubunzima bomzimba imihla ngemihla.
2.5. Ubungakanani bobunzima
Emva kokungeniswa kwe-OA, ukulinganiswa kwamandla okuthwala ubunzima bemilenze yangasemva kweerati zenziwa nge-incapacitance-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) njengoko kucwangcisiwe. Ukuhanjiswa kobunzima kumalungu angasemva kubalwa: ubunzima bokuthwala ubunzima (%)
