Osteoarthritis (OA) ni imwe mu ndwara zifata igihe kirekire zidakira zifata amagufwa yibasira abaturage bageze mu za bukuru barengeje imyaka 65 [
1]. Muri rusange, abarwayi ba OA basuzumwa na karitsiye yangiritse, synovium yaka umuriro, na chondrocytes yangirika, itera ububabare nububabare bwumubiri [
2]. Ububabare bwa rubagimpande buterwa ahanini no kwangirika kwa karitsiye mu ngingo hamwe no gutwikwa, kandi iyo karitsiye yangiritse cyane amagufwa arashobora kugongana bitera ububabare butihanganirwa ndetse nububabare bwumubiri [
3]. Uruhare rwabunzi batera ibimenyetso nibimenyetso nkububabare, kubyimba, no gukomera kwingingo byanditse neza. Mu barwayi ba OA, cytokine ikongora, itera isuri ya karitsiye hamwe namagufa ya subchondral iboneka mumazi ya synovial [
4]. Ibibazo bibiri by'ingenzi abarwayi ba OA bafite muri rusange ni ububabare no gutwika synovial. Kubwibyo intego yibanze yubuvuzi bwa OA nubu ni ukugabanya ububabare no gutwikwa. [
5]. Nubwo imiti ya OA iboneka, harimo imiti itari steroidal na steroidal, byagaragaye ko ifite akamaro mu kugabanya ububabare n’umuriro, gukoresha igihe kirekire iyi miti bigira ingaruka zikomeye ku buzima nk’umutima, imitsi, gastro-amara, ndetse n’imikorere mibi y’impyiko [
6]. Niyo mpamvu, imiti ikora neza ifite ingaruka nkeya igomba gutezwa imbere yo kuvura osteoarthritis.
Ibicuruzwa byubuzima bisanzwe bigenda byamamara kubera umutekano kandi byoroshye kuboneka [
7]. Imiti gakondo ya koreya yerekanye akamaro ko kurwanya indwara nyinshi zitera, harimo na rubagimpande [
8]. Aucklandia lappa DC. azwiho imiti yubuvuzi, nko kongera umuvuduko wa qi kugirango ugabanye ububabare no koroshya igifu, kandi wakoreshejwe gakondo nka analgesic naturel [
9]. Raporo zabanjirije iyi zerekana ko A. lappa ifite anti-inflammatory [
10,
11], analgesic [
12], anticancer [
13], na gastroprotective [
14] ingaruka. Ibikorwa bitandukanye byibinyabuzima bya A. lappa biterwa nibintu byingenzi bikora: costunolide, lahydone dehydrocostus, dihydrocostunolide, costuslactone, α-costol, saussurea lactone na costuslactone [
15]. Ubushakashatsi bwakozwe mbere buvuga ko costunolide yerekanaga imiti igabanya ubukana muri lipopolysaccharide (LPS), itera macrophage binyuze mu mabwiriza ya NF-kB hamwe n’ubushyuhe bwa poroteyine.
16,
17]. Ariko, nta bushakashatsi bwigeze bukora ubushakashatsi kubikorwa bya A. lappa yo kuvura OA. Ubu bushakashatsi bwakoze ubushakashatsi ku ngaruka zo kuvura A. lappa kurwanya OA ukoresheje (monosodium-iodoacetate) MIA hamwe na acide acike iterwa na moderi yimbeba.
Monosodium-iodoacetate (MIA) ikoreshwa cyane mu kubyara imyitwarire myinshi y'ububabare hamwe na patrophysiologique ya OA mu nyamaswa [
18,
19,
20]. Iyo MIA yatewe mu mavi, MIA itesha agaciro metabolisme ya chondrocyte kandi igatera ibimenyetso byerekana umuriro, nka karitsiye hamwe nisuri yamagufwa ya subchondral, ibimenyetso byingenzi bya OA [
18]. Igisubizo cyanditse cyatewe na acide acetike gifatwa nkikigereranyo cyububabare bwa periferique ku nyamaswa aho ububabare bwakongejwe bushobora gupimwa ku bwinshi.
19]. Imbeba macrophage selile umurongo, RAW264.7, ikoreshwa cyane mukwiga ibisubizo bya selile kumuriro. Iyo ukorana na LPS, macrophage ya RAW264 ikora inzira yumuriro kandi ikarekura abahuza benshi, nka TNF-α, COX-2, IL-1β, iNOS, na IL-6 [
20]. Ubu bushakashatsi bwasuzumye ingaruka zo kurwanya nociceptive na anti-inflammatory ya A. lappa irwanya OA mu cyitegererezo cy’inyamanswa ya MIA, icyitegererezo cy’inyamanswa iterwa na acide, na selile RAW264.7.
2. Ibikoresho nuburyo bwiza
2.1. Ibikoresho by'ibihingwa
Umuzi wumye wa A. lappa DC. yakoreshejwe mu igeragezwa yaguzwe muri Epulip Pharmaceutical Co., Ltd., (Seoul, Koreya). Yagaragajwe na Prof. Donghun Lee, Umuyobozi w’imiti y’imiti y’ibyatsi, Col. w’ubuvuzi bwa Koreya, kaminuza ya Gachon, kandi nimero yerekana inyemezabuguzi yashyizwe mu 18060301.
2.2. HPLC Isesengura rya A. lappa Ikuramo
A. lappa yakuweho hakoreshejwe ibikoresho byo kugaruka (amazi yatoboye, 3 h kuri 100 ° C). Igisubizo cyakuweho cyayungurujwe kandi cyegeranye hifashishijwe moteri yumuvuduko muke. A. ikuramo rya lappa ryagize umusaruro wa 44,69% nyuma yo gukama-gukama munsi ya −80 ° C. Isesengura rya Chromatografique ya A. lappa ryakozwe na HPLC ihujwe hakoreshejwe sisitemu ya 1260 InfinityⅡ HPLC (Agilent, Pal Alto, CA, USA). Gutandukanya chromatic, EclipseXDB C18 inkingi (4,6 × 250 mm, 5 µm, Agilent) yakoreshejwe kuri 35 ° C. Igiteranyo cya mg 100 cyikigereranyo cyavanze muri mL 10 ya 50% methanol hanyuma ikabyara iminota 10. Ingero zayungurujwe hamwe na filteri ya siringi (Amazi Corp., Milford, MA, USA) ya 0.45 mm. Igice cya mobile kigizwe na 0.1% acide fosifori (A) na acetonitrile (B) hanyuma inkingi yavuzwe kuburyo bukurikira: 0-60 min, 0%; Iminota 60-65, 100%; Iminota 65-67, 100%; 67-72 min, 0% solvent B hamwe nigipimo cya 1.0 mL / min. Amazi yagaragaye kuri 210 nm akoresheje inshinge ya 10 μL. Isesengura ryakozwe muri bitatu.
2.3. Imiturire ninyamaswa
Imbeba Zigitsina gabo - Dawley (SD) zifite ibyumweru 5 nimbeba za ICR zumugabo zifite ibyumweru 6 zaguzwe muri Samtako Bio Korea (Gyeonggi-do, Koreya). Amatungo yabitswe mucyumba akoresheje ubushyuhe buhoraho (22 ± 2 ° C) nubushuhe (55 ± 10%) hamwe n’umucyo / umwijima wa 12/12 h. Inyamaswa zari zimenyereye imiterere mugihe kirenze icyumweru mbere yuko ubushakashatsi butangira. Amatungo yari afite ad libitum itanga ibiryo n'amazi. Amategeko agenga imyitwarire yubu yo kwita ku nyamaswa no gufata neza muri kaminuza ya Gachon (GIACUC-R2019003) yakurikijwe cyane muburyo bwose bwo kugerageza inyamaswa. Ubushakashatsi bwateguwe nubushakashatsi buhumye kandi bubangikanye. Twakurikije uburyo bwa euthanasiya dukurikije umurongo ngenderwaho wa komite ishinzwe imyitwarire yinyamaswa.
2.4. Gutera inshinge no kuvura
Imbeba zatandukanijwe muburyo butandukanye mumatsinda 4, arizo sham, kugenzura, indomethacine, na A. lappa. Kuba anesthete hamwe na 2% isofluorane O2 ivanze, imbeba zatewe inshinge 50 μL za MIA (40 mg / m; Sigma-Aldrich, St. Ubuvuzi bwakozwe nkibi bikurikira: kugenzura amatsinda ya sham byakomezaga gusa nimirire yibanze ya AIN-93G. Gusa, itsinda rya indomethacin ryahawe indomethacine (3 mg / kg) yinjijwe mu ndyo ya AIN-93G naho itsinda rya A. lappa 300 mg / kg ryahawe indyo ya AIN-93G yongerewe na A. lappa (300 mg / kg). Ubuvuzi bwakomeje iminsi 24 uhereye umunsi OA yinjiriye ku gipimo cya 15-17 g ku buremere bw'umubiri wa 190-210 g buri munsi.
2.5. Ibipimo byo gupima ibiro
Nyuma yo kwinjizwa kwa OA, gupima ubushobozi bwo gupima uburemere bwinyuma yinyuma yimbeba byakozwe hamwe nubushobozi buke-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) nkuko byari byateganijwe. Ikwirakwizwa ryibiro ku ngingo zinyuma ryabazwe: ubushobozi bwo gutwara ibiro (%)
