Osteoarthritis (OA) yog ib qho ntawm cov kab mob ntev ntev uas cuam tshuam rau cov neeg laus tshaj 65 xyoo [
1]. Feem ntau, cov neeg mob OA raug kuaj pom tias muaj cov pob txha mos uas puas lawm, cov synovium o, thiab cov chondrocytes uas lwj, uas ua rau mob thiab ntxhov siab rau lub cev [
2]. Mob pob qij txha feem ntau yog tshwm sim los ntawm kev puas tsuaj ntawm cov pob txha mos hauv cov pob qij txha los ntawm kev o, thiab thaum cov pob txha mos raug puas tsuaj loj heev, cov pob txha tuaj yeem sib tsoo ua rau mob hnyav thiab ua rau lub cev tsis muaj zog [
3]. Kev koom tes ntawm cov neeg sawv cev rau kev o nrog cov tsos mob xws li mob, o, thiab mob pob qij txha tau sau tseg zoo. Hauv cov neeg mob OA, cov cytokines o, uas ua rau cov pob txha mos thiab cov pob txha subchondral lwj tau pom nyob rau hauv cov kua synovial [
4]. Ob qho kev tsis txaus siab loj uas cov neeg mob OA feem ntau muaj yog qhov mob thiab o ntawm cov pob qij txha. Yog li ntawd, lub hom phiaj tseem ceeb ntawm kev kho mob OA tam sim no yog kom txo qhov mob thiab o.
5]. Txawm hais tias cov kev kho mob OA muaj, suav nrog cov tshuaj tsis yog steroidal thiab cov tshuaj steroidal, tau ua pov thawj tias muaj txiaj ntsig zoo hauv kev txo qhov mob thiab o, kev siv cov tshuaj no mus sij hawm ntev muaj cov teeb meem loj rau kev noj qab haus huv xws li kev ua haujlwm ntawm lub plawv, txoj hnyuv, thiab lub raum [
6Yog li ntawd, yuav tsum tsim ib yam tshuaj zoo dua uas muaj tsawg dua cov kev mob tshwm sim rau kev kho mob osteoarthritis.
Cov khoom noj khoom haus ntuj tsim tau nrov zuj zus vim muaj kev nyab xeeb thiab yooj yim nrhiav tau [
7]. Cov tshuaj Korean ib txwm muaj tau ua pov thawj tias muaj txiaj ntsig zoo rau ntau yam kab mob o, suav nrog mob pob qij txha [
8]. Aucklandia lappa DC. paub txog nws cov khoom siv tshuaj, xws li txhim kho kev ncig ntawm qi rau kev txo qhov mob thiab so lub plab, thiab tau siv ib txwm ua tshuaj analgesic ntuj [
9]. Cov ntawv ceeb toom yav dhau los qhia tias A. lappa muaj cov tshuaj tiv thaiv kev o [
10,
11], tshuaj loog mob [
12], tshuaj tiv thaiv kab mob qog noj ntshav [
13], thiab tiv thaiv kab mob plab [
14] cov teebmeem. Cov haujlwm ntawm A. lappa ntau yam yog tshwm sim los ntawm nws cov tshuaj tseem ceeb: costunolide, dehydrocostus lactone, dihydrocostunolide, costuslactone, α-costol, saussurea lactone thiab costuslactone [
15]. Cov kev tshawb fawb ua ntej tau hais tias costunolide tau qhia txog cov khoom tiv thaiv kev o hauv lipopolysaccharide (LPS), uas ua rau cov macrophages los ntawm kev tswj hwm ntawm NF-kB thiab txoj kev ua kom sov kub [
16,
17]. Txawm li cas los xij, tsis muaj kev tshawb fawb twg tau tshawb nrhiav txog cov haujlwm ntawm A. lappa rau kev kho mob OA. Cov kev tshawb fawb tam sim no tau tshawb nrhiav cov txiaj ntsig kho mob ntawm A. lappa tawm tsam OA siv (monosodium-iodoacetate) MIA thiab acetic acid-induced rodent models.
Monosodium-iodoacetate (MIA) yog siv los tsim ntau yam kev mob thiab cov yam ntxwv pathophysiological ntawm OA hauv tsiaj txhu [
18,
19,
20]. Thaum txhaj rau hauv cov pob qij txha hauv caug, MIA ua rau lub cev tsis ua haujlwm ntawm chondrocyte thiab ua rau muaj kev o thiab cov tsos mob ntawm kev o, xws li cov pob txha mos thiab cov pob txha subchondral erosion, cov tsos mob tseem ceeb ntawm OA [
18]. Kev teb rov qab los ntawm acetic acid yog suav tias yog kev sim ua qhov mob ntawm cov tsiaj uas qhov mob o tuaj yeem ntsuas tau ntau yam [
19]. Cov kab mob macrophage nas, RAW264.7, yog siv los kawm txog cov lus teb ntawm tes rau kev o. Thaum ua haujlwm nrog LPS, RAW264 macrophages ua rau cov kab mob o thiab tso tawm ntau yam tshuaj o, xws li TNF-α, COX-2, IL-1β, iNOS, thiab IL-6 [
20]. Txoj kev tshawb fawb no tau soj ntsuam cov teebmeem tiv thaiv kev mob thiab kev mob ntawm A. lappa tawm tsam OA hauv MIA tsiaj qauv, acetic acid-induced tsiaj qauv, thiab LPS-activated RAW264.7 hlwb.
2. Cov Khoom Siv thiab Cov Txheej Txheem
2.1. Cov Khoom Siv Cog Qoob Loo
Cov cag qhuav ntawm A. lappa DC. siv rau hauv kev sim tau los ntawm Epulip Pharmaceutical Co., Ltd., (Seoul, Kauslim Teb). Nws tau txheeb xyuas los ntawm Prof. Donghun Lee, Dept. ntawm Herbal pharmacology, Col. ntawm Korean Medicine, Gachon University, thiab tus lej qauv voucher tau muab tso ua 18060301.
2.2. Kev Tshuaj Xyuas HPLC ntawm A. lappa Extract
A. lappa tau rho tawm siv lub tshuab reflux (dej distilled, 3 teev ntawm 100 ° C). Cov kua uas tau rho tawm tau lim thiab condensed siv lub tshuab evaporator qis-siab. A. lappa extract muaj cov txiaj ntsig ntawm 44.69% tom qab khov-qhuav hauv qab −80 ° C. Kev tshuaj xyuas Chromatographic ntawm A. lappa tau ua nrog HPLC txuas nrog siv 1260 InfinityⅡ HPLC-system (Agilent, Pal Alto, CA, USA). Rau kev sib cais chromatic, EclipseXDB C18 kem (4.6 × 250 mm, 5 µm, Agilent) tau siv ntawm 35 ° C. Tag nrho ntawm 100 mg ntawm cov qauv tau diluted hauv 10 mL ntawm 50% methanol thiab sonicated rau 10 feeb. Cov qauv tau lim nrog lub lim dej syringe (Waters Corp., Milford, MA, USA) ntawm 0.45 μm. Cov khoom sib xyaw ua ke ntawm theem mobile yog 0.1% phosphoric acid (A) thiab acetonitrile (B) thiab kem tau eluted raws li nram no: 0–60 feeb, 0%; 60–65 feeb, 100%; 65–67 feeb, 100%; 67–72 feeb, 0% solvent B nrog tus nqi ntws ntawm 1.0 mL / feeb. Cov dej khib nyiab tau pom ntawm 210 nm siv qhov ntim txhaj tshuaj ntawm 10 μL. Kev tshuaj xyuas tau ua tiav hauv triplicate.
2.3. Kev Tswj Xyuas thiab Kev Tswj Xyuas Tsiaj
Cov nas txiv neej Sprague-Dawley (SD) hnub nyoog 5 lub lis piam thiab cov nas txiv neej ICR hnub nyoog 6 lub lis piam tau yuav los ntawm Samtako Bio Korea (Gyeonggi-do, Kauslim Teb). Cov tsiaj tau khaws cia rau hauv ib chav siv qhov kub thiab txias tas li (22 ± 2 °C) thiab av noo (55 ± 10%) thiab lub voj voog kaj / tsaus ntuj ntawm 12/12 teev. Cov tsiaj tau paub txog qhov xwm txheej ntau dua ib lub lim tiam ua ntej qhov kev sim pib. Cov tsiaj muaj kev pub zaub mov thiab dej ad libitum. Cov cai tam sim no rau kev saib xyuas thiab kev tuav tsiaj ntawm Gachon University (GIACUC-R2019003) tau ua raws li nruj me ntsis hauv txhua txoj kev sim tsiaj. Kev tshawb fawb tau tsim los ntawm tus kws tshawb nrhiav-dig muag thiab kev sim sib luag. Peb tau ua raws li txoj kev euthanasia raws li cov lus qhia ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Tsiaj.
2.4. Kev Txhaj Tshuaj MIA thiab Kev Kho Mob
Cov nas tsuag tau raug faib ua 4 pawg, uas yog sham, tswj, indomethacin, thiab A. lappa. Thaum muab tshuaj loog nrog 2% isofluorane O2 sib tov, cov nas tsuag tau txhaj tshuaj siv 50 μL ntawm MIA (40 mg / m2; Sigma-Aldrich, St. Louis, MO, USA) rau hauv cov pob qij txha hauv caug kom ua rau muaj kev sim OA. Cov kev kho mob tau ua raws li hauv qab no: pawg tswj thiab sham tau tswj hwm tsuas yog nrog AIN-93G zaub mov yooj yim. Tsuas yog, pawg indomethacin tau muab indomethacin (3 mg / kg) uas tau koom nrog AIN-93G zaub mov thiab pawg A. lappa 300 mg / kg tau muab rau AIN-93G zaub mov ntxiv nrog A. lappa (300 mg / kg). Cov kev kho mob tau txuas ntxiv mus rau 24 hnub txij li hnub ntawm OA induction ntawm tus nqi ntawm 15-17 g ib 190-210 g lub cev hnyav txhua hnub.
2.5. Kev Ntsuas Qhov Hnyav
Tom qab OA induction, kev ntsuas qhov hnyav ntawm cov ceg tom qab ntawm cov nas tau ua tiav nrog incapacitance-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) raws li tau teem sijhawm. Kev faib qhov hnyav ntawm cov ceg tom qab tau suav: qhov hnyav muaj peev xwm (%)
