Kim, Jaeyong
(Jeonnam Institute of Natural Resources Research)
,
Yang, Siyoung
(Korea Research Institute of Bioscience and Biotechnology)
,
Choi, Chul-yung
(Jeonnam Institute of Natural Resources Research)
In this study, the anti-osteoarthritis effects of Cynanchum wilfordii, Phlomis umbrosa, and Angelica gigas extract (CPAE), observed and confirmed in previously clinical studies were further investigated by in vitro and in vivo studies. Anabolic biomarkers related to healthy cartilage maintenance, su...
In this study, the anti-osteoarthritis effects of Cynanchum wilfordii, Phlomis umbrosa, and Angelica gigas extract (CPAE), observed and confirmed in previously clinical studies were further investigated by in vitro and in vivo studies. Anabolic biomarkers related to healthy cartilage maintenance, such as aggrecan, type II collagen ${\alpha}$-1 (Col2a1), sex determining region Y-box-9 (Sox-9), and catabolic biomarkers related to osteoarthritis, such as cyclooxygenase-2 (Cox-2), matrix metalloproteinase-13 (Mmp13), and nuclear factor kappa-light-chain-enhancer of activated B cells ($Nf{\kappa}b$), were evaluated by quantitative reverse transcriptase polymerase chain reaction and reporter gene assay. In vitro study results showed significant changes in both anabolic and catabolic biomarkers. For anabolic factors, significant changes in the level of aggrecan (P<0.05), Col2a1 (P<0.05), and Sox-9 (P<0.01) activation were shown after treatment of cartilage cells with CPAE (50 ng/mL) with similar efficacy compared to insulin growth factor, the positive control (100 ng/mL). For catabolic factors, significant changes in the inhibition activity of Cox-2 (P<0.05), Mmp13 (P<0.01), and $Nf{\kappa}b$ (P<0.05) were shown for CPAE (50 ng/mL) with similar efficacy compared to Celecoxib, the positive control ($10{\mu}M$). In the in vivo carrageenan-induced paw edema model study results showed that CPAE-treated groups (100 mg/kg) and Celecoxib-treated groups (60 mg/kg) showed comparably significant efficacy of inhibition by 37.1% and 52.1%, respectively. Furthermore, CPAE (200 mg/kg) showed similar effect to Celecoxib (60 mg/kg) with an inhibition rate of 54.3%. This result confirms that CPAE effectively inhibited the inflammation-induced osteoarthritis symptoms.
In this study, the anti-osteoarthritis effects of Cynanchum wilfordii, Phlomis umbrosa, and Angelica gigas extract (CPAE), observed and confirmed in previously clinical studies were further investigated by in vitro and in vivo studies. Anabolic biomarkers related to healthy cartilage maintenance, such as aggrecan, type II collagen ${\alpha}$-1 (Col2a1), sex determining region Y-box-9 (Sox-9), and catabolic biomarkers related to osteoarthritis, such as cyclooxygenase-2 (Cox-2), matrix metalloproteinase-13 (Mmp13), and nuclear factor kappa-light-chain-enhancer of activated B cells ($Nf{\kappa}b$), were evaluated by quantitative reverse transcriptase polymerase chain reaction and reporter gene assay. In vitro study results showed significant changes in both anabolic and catabolic biomarkers. For anabolic factors, significant changes in the level of aggrecan (P<0.05), Col2a1 (P<0.05), and Sox-9 (P<0.01) activation were shown after treatment of cartilage cells with CPAE (50 ng/mL) with similar efficacy compared to insulin growth factor, the positive control (100 ng/mL). For catabolic factors, significant changes in the inhibition activity of Cox-2 (P<0.05), Mmp13 (P<0.01), and $Nf{\kappa}b$ (P<0.05) were shown for CPAE (50 ng/mL) with similar efficacy compared to Celecoxib, the positive control ($10{\mu}M$). In the in vivo carrageenan-induced paw edema model study results showed that CPAE-treated groups (100 mg/kg) and Celecoxib-treated groups (60 mg/kg) showed comparably significant efficacy of inhibition by 37.1% and 52.1%, respectively. Furthermore, CPAE (200 mg/kg) showed similar effect to Celecoxib (60 mg/kg) with an inhibition rate of 54.3%. This result confirms that CPAE effectively inhibited the inflammation-induced osteoarthritis symptoms.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
In addition, a promoter analysis was also performed to further confirm the regulation of Sox-9 activation as well as its associated cofactors by CPAE. Treatment with CPAE at different concentrations increased Sox-9 activation.
In this study, IGF was used as positive control for testing the increase of the anabolic factor, and Celecoxib was used as positive control for testing the inhibition of the expression of catabolic factor and in in vivo test. IGF was reported as the major regulating factor of cartilage proteoglycan synthesis in human synovial fluid (20), and Celecoxib that selectively inhibit COX-2 might block inflammation, pain and fever while reducing the side effects (21).
In addition, there are several animal models for evaluating arthritis related clinical biomarkers, and one of the most popular methods involves the induction of foot, ear, or knee edema after treatment with the test agent, and then measurements are subsequently taken using a plethysmometer (17). In this study, the carrageenan-induced rat paw edema model was used to measure the degree of edema inhibition by CPAE and, thereby, verify its efficacy in treating arthritis (18,19).
One hour before the carrageenan injection, Celecoxib (60 mg/kg) as positive control was given orally, while the CPAE treatment groups were administered 50, 100, and 200 mg/kg for 4 days, and 1 h following the treatment on day 4, 1% (w/v) carrageenan (100 μL) was injected into the right hind paw of each rat.
The following conditions and primers were used for the qRT-PCR: Col2a1 (annealing temperature 60℃), sense 5’-CAC ACT GGT AAG TGG GGC AAG A-3’ and antisense 5’-GGA TTG TGT TGT TTC AGG GTT CG-3’; Sox-9 (annealing temperature 55oC), sense 5’-CAT CAG CAG CAC CGC ACC CA-3’ and antisense 5’-CGG GTG ATG GGC GGG TAG GA-3’; aggrecan (annealing temperature 60℃), sense 5’-GAA GAC GAC ATC ACC ATC CAG-3’ and antisense 5’-CTG TCT TTG TCA CCC ACA CAT G-3’; Mmp-13 (annealing temperature 55oC), sense 5’-TGA TGG ACC TTC TGG TCT TCT GG-3’ and antisense 5’-CAT CCA CAT GGT TGG GAA GTT CT-3’; Cox-2 (annealing temperature 62oC), sense 5’-GGT CTG GTG CCT GGT CTG ATG AT-3’ and antisense 5’-GTC CTT TCA AGG GAG AAT GGT GC-3’.
This study was designed to evaluate the inhibition of arthritis after the administration of CPAE by evaluating its effects on the maintenance of cartilage health and osteoarthritis using a cartilage cell model. Furthermore, a carrageenan-induced rat paw edema model was used to evaluate the improvement of inflammation-induced symptoms of arthritis with treatment of CPAE.
대상 데이터
The 5∼6-week-old male Sprague-Dawley (SD) rats used in this study were purchased from Samtako (Osan, Korea) and weighed 180∼200 g.
데이터처리
*P<0.05 and **P<0.01 compared with control, as analyzed by Student’s t-test.
*P<0.05 and **P<0.01 compared with interleukin-1β (IL-1β), as analyzed by Student’s t-test.
The results are expressed as mean±standard error of the mean (SEM), and the Student’s t-test was used for the statistical analysis and P<0.05 and P<0.01 where considered statistically significant.
Values are mean±standard error of the mean, *P<0.05 and **P<0.01 compared with interleukin-1β (IL-1β), as analyzed by Student’s t-test.
성능/효과
After administrating 50, 100, and 200 mg/kg of CPAE for 4 days followed by the induction of acute inflammation with carrageenan, the hourly change in the level of edema was measured. CPAE significantly reduced the carrageenan-induced paw edema at concentrations above 100 mg/kg compared to Control group, and the inhibition rate was highest at 200 mg/kg of CPAE (Fig.
3). The results revealed that CPAE and each of the three root extracts inhibited Cox-2 activation. In particular, CPAE at a concentration of 50 ng/mL showed a similar inhibitory efficacy to that of the control compound, Celecoxib at 10 μM.
1B). The results revealed that treatment with CPAE induced a higher activation of the anabolic factors than each of the individual root extracts did, suggesting that the combination of the three roots produced synergistic effects. Furthermore, CPAE at a concentration of 50 ng/mL showed similar activation to that of the control compound, IGF at 100 ng/mL.
When a foreign substance invades the human body, normal immune responses would produce vasoactive substances, such as histamine, serotonin, bradykinin, prostaglandin, and leukotriene that increase vascular membrane permeability leading to edema, and this inflammation could be the cause for arthritis. This study shows that CPAE has anti-inflammatory effects to inhibit edema, and EstroG-100 could prevent and inhibit arthritis caused by acute inflammation.
참고문헌 (26)
1 Elders MJ 2000 The increasing impact of arthritis on public health J Rheumatol Suppl 60 6 8 11032095
2 Høegh-Andersen P Tankó LB Andersen TL undberg CV Mo JA Heegaard AM Delaissé JM Christgau S 2004 Ovariectomized rats as a model of postmenopausal osteoarthritis: validation and application Arthritis Res Ther 6 R169 R180 10.1186/ar1152 400436 --> 15059281
4 Hart DJ Doyle DV Spector TD 1999 Incidence and risk factors for radiographic knee osteoarthritis in middle-aged women: the Chingford Study Arthritis Rheum 42 17 24 10.1002/1529-0131(199901)42:1 3.0.CO;2-E 9920009
5 Nevitt MC Cummings SR Lane NE Hochberg MC Scott JC Pressman AR Genant HK Cauley JA 1996 Association of estrogen replacement therapy with the risk of osteoarthritis of the hip in elderly white women Arch Intern Med 156 2073 2080 10.1001/archinte.1996.00440170081009 8862099
8 Hannan MT Felson DT Anderson JJ Naimark A Kannel WB 1990 Estrogen use and radiographic osteoarthritis of the knee in women Arthritis Rheum 33 525 532 10.1002/art.1780330410 2328031
9 Rossouw JE Anderson GL Prentice RL LaCroix AZ Kooperberg C Stefanick ML Jackson RD Beresford SA Howard BV Johnson KC Kotchen JM Ockene J Writing Group for the Women’s Health Initiative Investigators 2002 Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results from the Women’s Health Initiative randomized controlled trial JAMA 288 321 333 10.1001/jama.288.3.321 12117397
10 Collaborative Group on Epidemiological Studies of Ovarian Cancer Beral V Gaitskell K Hermon C Moser K Reeves G Peto R 2015 Menopausal hormone use and ovarian cancer risk: individual participant meta-analysis of 52 epidemiological studies Lancet 385 1835 1842 10.1016/S0140-6736(14)61687-1 4427760 --> 25684585
11 Lee KH Lee DJ Kim SM Je SH Kim EK Han HS Han IK 2005 Evaluation of effectiveness and safety of natural plants extract (Estromon ® ) on perimenopausal women for 1 year J Menopausal Med 11 16 26
12 Chang A Kwak BY Yi K Kim JS 2012 The effect of herbal extract (EstroG-100) on pre-, peri- and post-menopausal women: a randomized double-blind, placebo-controlled study Phytother Res 26 510 516 10.1002/ptr.3597 21887807
13 Fraser A Fearon U Billinghurst RC Ionescu M Reece R Barwick T Emery P Poole AR Veale DJ 2003 Turnover of type II collagen and aggrecan in cartilage matrix at the onset of inflammatory arthritis in humans: relationship to mediators of systemic and local inflammation Arthritis Rheum 48 3085 3095 10.1002/art.11331 14613270
14 Kim KI Park YS Im GI 2013 Changes in the epigenetic status of the SOX-9 promoter in human osteoarthritic cartilage J Bone Miner Res 28 1050 1060 10.1002/jbmr.1843 23225119
15 Yang S Kim J Ryu JH Oh H Chun CH Kim BJ Min BH Chun JS 2010 Hypoxia-inducible factor-2α is a catabolic regulator of osteoarthritic cartilage destruction Nat Med 16 687 693 10.1038/nm.2153 20495569
16 Futaki N Takahashi S Yokoyama M Arai I Higuchi S Otomo S 1994 NS-398, a new anti-inflammatory agent, selectively inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity in vitro Prostaglandins 47 55 59 10.1016/0090-6980(94)90074-4 8140262
19 Yang Y Leech M Hutchinson P Holdsworth SR Morand EF 1997 Antiinflammatory effect of lipocortin 1 in experimental arthritis Inflammation 21 583 596 10.1023/A:1027330021479 9429906
20 Schalkwijk J Joosten LA van den Berg WB van Wyk JJ van de Putte LB 1989 Insulin-like growth factor stimulation of chondrocyte proteoglycan synthesis by human synovial fluid Arthritis Rheum 32 66 71 10.1002/anr.1780320111 2912464
21 Goldenberg MM 1999 Celecoxib, a selective cyclooxygenase-2 inhibitor for the treatment of rheumatoid arthritis and osteoarthritis Clin Ther 21 1497 1513 10.1016/S0149-2918(00)80005-3 10509845
22 Koo HJ Sohn EH Pyo S Woo HG Park DW Ham YM Jang SA Park SY Kang SC 2015 An ethanol root extract of Cynanchum wilfordii containing acetophenones suppresses the expression of VCAM-1 and ICAM-1 in TNF-α-stimulated human aortic smooth muscle cells through the NF-κB pathway Int J Mol Med 35 915 924 4356471 --> 25716870
23 Yang SB Lee SM Park JH Lee TH Baek NI Park HJ Lee H Kim J 2014 Cynandione A from Cynanchum wilfordii attenuates the production of inflammatory mediators in LPS-induced BV-2 microglial cells via NF-κB inactivation Biol Pharm Bull 37 1390 1396 10.1248/bpb.b13-00939 25087960
24 Shang X Wang J Li M Miao X Pan H Yang Y Wang Y 2011 Antinociceptive and anti-inflammatory activities of Phlomis umbrosa Turcz extract Fitoterapia 82 716 721 10.1016/j.fitote.2011.03.001 21406221
25 Kim JH Jeong JH Jeon ST Kim H Ock J Suk K Kim SI Song KS Lee WH 2006 Decursin inhibits induction of inflammatory mediators by blocking nuclear factor-κB activation in macrophages Mol Pharmacol 69 1783 1789 10.1124/mol.105.021048 16510559
26 Rim HK Cho W Sung SH Lee KT 2016 Nodakenin suppresses lipopolysaccharide-induced inflammatory responses in macrophage cells by inhibiting tumor necrosis factor receptor-associated factor 6 and nuclear factor-κB pathways and protects mice from lethal endotoxin shock J Pharmacol Exp Ther 342 654 664 10.1124/jpet.112.194613
※ AI-Helper는 부적절한 답변을 할 수 있습니다.