Suppression of inflammatory mediators and matrix metalloproteinase (MMP)-13 by Morus alba stem extract and oxyresveratrol in RAW 264.7 cells and C28/I2 human chondrocytes

10.1016/j.jtcme.2019.03.006

Thidarat Wongwat；Kanyarat Srihaphon；Chetsadaporn Pitaksutheepong；Worawan Boonyo；Tasana Pitaksuteepong

Mulberry stem ； Oxyresveratrol ； Chondrocytes ； Chondroprotective ； Inflammation

Journal of Traditional and Complementary Medicine

10卷2期（2020 / 03 / 01）

132 - 140

英文

This study aimed to investigate the effects of Morus alba stem extract (MSE) and oxyresveratrol on the suppression of pro-inflammatory mediators in LPS-stimulated RAW 264.7 macrophages and IL-1β-stimulated C28/I2 human chondrocyte cell line. The chondroprotective effect was also investigated using the chondrocyte cell line. First, MSE was prepared and analyzed for the amount of oxyresveratrol. The anti-inflammatory effects of MSE at various concentrations were evaluated through the inhibition of nitric oxide (NO), prostaglandin (PG)-E_2 and cyclooxygenase (COX)-2 production. Oxyresveratrol at the equivalent amount found in the extract was investigated in the same manner. The chondroprotective effect was investigated through the suppression of MMP-13 production. The results showed that oxy-resveratrol content in MSE was 15%. In RAW 264.7 cells, MSE (5-50 μg/mL) could inhibit the NO (24-30%) and PGE_2 (11-82%) production. Oxyresveratrol at 0.75 and 7.5 mg/μL could suppress NO and also inhibited PGE_2 but at only at high concentration. In the chondrocyte cell line, MSE at 5-100 mg/μL significantly decreased the PGE_2 and COX-2 production by 44-93% and 17-65%, respectively. Again, oxyresveratrol at both concentrations could significantly inhibit PGE_2 production by 50-92% but it inhibited COX-2 only at high concentration. In addition, MSE and oxyresveratrol was shown to significantly inhibit MMP-13 production by 14-57% and 16-56%, depending on their concentrations. The MSE demonstrates the potential to be used as an alternative treatment for reducing inflammation and preventing cartilage degradation. Its component, oxyresveratrol, may exert these effects to some extent.

1. Huang, HP,Ou, TT,Wang, CJ(2013).Mulberry（桑葚子Sang Shèn Zǐ）and its bioactive compounds, the chemoprevention effects and molecular mechanisms in vitro and in vivo.J Tradit Complement Med,3(1),7-15.
   連結：
2. Pan, MH,Chiou, YS,Tsai, ML,Ho, CT(2011).Anti-inflammatory activity of traditional Chinese medicinal herbs.J Tradit Complement Med,1(1),8-24.
   連結：
3. Abramson, SB(1999).The role of COX-2 produced by cartilage in arthritis.Osteoarthritis Cartilage,7(4),380-381.
4. Al-Kofahi, M,Becker, F,Gavins, FN(2015).IL-1β reduces tonic contraction of mesenteric lymphatic muscle cells, with the involvement of cyclooxygenase-2 and prostaglandin E2.Br J Pharmacol,172(16),4038-4051.
5. Bensky, D,Gamble, A.(1993).Chinese Herbal Medicine: Materia Medica.Washington:Eastland Press.
6. Burrage, PS,Mix, KS,Brinckerhoff, CE(2006).Matrix metalloproteinases: role in arthritis.Front Biosci,11,529-543.
7. Chen, P,Zheng, F,Zhang, Y,Gao, F,Chen, Y,Shi, G(2016).Ethnobotanical study of medicinal plants on arthritis used by Chaoshan in Guangdong, China.Med Chem.,6(12),715-723.
8. Chung, KO,Kim, BY,Lee, MH(2003).In-vitro and in-vivo anti-inflammatory effect of oxyresveratrol from Morus alba L.J Pharm Pharmacol,55(12),1695-1700.
9. Eo, HJ,Park, JH,Park, GH(2014).Anti-inflammatory and anti-cancer activity of mulberry (Morus alba L.) root bark.BMC Complement Altern Med,14,200.
10. Fernandes, JC,Pelletier, JM,Pelletier, JP(2002).The role of cytokines in osteoarthritis pathophysiology.Biorheology,39(1-2),237-246.
11. Glyn-Jones, S,Palmer, AJ,Agricola, R(2015).Osteoarthritis.Lancet,386,376-387.
12. Goldring, MB,Goldring, SR(2007).Osteoarthritis.J Cell Physiol,213(3),626-634.
13. Goldring, MB,Otero, M(2011).Inflammation in osteoarthritis.Curr Opin Rheumatol,23(5),471-478.
14. Kapoor, M,Mineau, F,Fahmi, H,Pelletier, JP,Martel-Pelletier, J(2012).Glucosamine sulfate reduces prostaglandin E2 production in osteoarthritic chondrocytes through inhibition of microsomal PGE synthase-1.J Rheumatol,39(3),635-644.
15. Kargman, S,Charleson, S,Cartwright, M(1996).Characterization of prostaglandin G/H synthase 1 and 2 in rat, dog, monkey, and human gastrointestinal tracts.Gastroenterology,111(2),445-454.
16. Katsube, T,Imawaka, N,Kawano, Y,Yamazaki, Y,Shiwaku, K,Yamane, Y(2006).Antioxidant flavonol glycosides in mulberry (Morus alba L.) leaves isolated based on LDL antioxidant activity.Food Chem,97(1),25-31.
17. Khunakornvichaya, A,Lekmeechai, S,Pham, PP(2016).Morus alba L. stem extract attenuates pain and articular cartilage damage in the anterior cruciate ligament transection-Induced rat model of osteoarthritis.Pharmacology,98(5-6),209-216.
18. Kim, DC,Quang, TH,Oh, H,Kim, YC(2017).Steppogenin isolated from Cudrania tricuspidata shows antineuroinflammatory effects via NF-kB and MAPK pathways in LPS-stimulated BV2 and primary rat microglial cells.Molecules,22(12)
19. Koopman, WJ,Moreland, LW.(2005).Arthritis and Allied Conditions: A Textbook of Rheumatology.Philadelphia:Lippincott Williams & Wilkins.
20. Lee, HS,Kim, DH,Hong, JE,Lee, JY,Kim, EJ(2015).Oxyresveratrol suppresses lipopolysaccharide-induced inflammatory responses in murine macrophages.Hum Exp Toxicol,34(8),808-818.
21. Lee, JH,Ko, HJ,Woo, ER(2016).Moracin M inhibits airway inflammation by interrupting the JNK/C-Jun and NF-kB pathways in vitro and vivo.Eur J Pharmacol,783,64-72.
22. Lin, WW,Chen, BC,Hsu, YW,Lee, CM,Shyue, SK(1999).Modulation of inducible nitric oxide synthase induction by prostaglandin E2 in macrophages: distinct susceptibility in murine J774 and RAW 264.7 macrophages.Prostag Other Lipid Mediat,58(2-4),87-101.
23. Liu, FC,Hung, LF,Wu, WL(2010).Chondroprotective effects and mechanisms of resveratrol in advanced glycation end products-stimulated chondrocytes.Arthritis Res Ther,12(5),R167.
24. Mercurio, F,Manning, AM(1999).Multiple signals converging on NF-kB.Curr Opin Cell Biol,11(2),226-232.
25. O'Neill, LA,Lewis, GP(1989).Inhibitory effects of diclofenac and indomethacin on interleukin-1-induced changes in PGE2 release. A novel effect on free arachidonic acid levels in human synovial cells.Biochem Pharmacol,38(21),3707-3711.
26. Park, E,Lee, S,Lee, J,Kim, J(2013).Anti-inflammatory activity of mulberry leaf extract through inhibition of NF-kB.J Funct Foods,5(1),178-186.
27. Reboul, P,Pelletier, JP,Tardif, G,Cloutier, JM,Martel-Pelletier, J(1996).The new collagenase, collagenase-3, is expressed and synthesized by human chondrocytes but not by synoviocytes.J Clin Investig,97(9),2011-2019.
28. Rivière, C,Krisa, S,Péchamat, L(2014).Polyphenols from the stems of Morus alba and their inhibitory activity against nitric oxide production by lipopolysaccharide-activated microglia.Fitoterapia,97,253-260.
29. Shakibaei, M,Csaki, C,Nebrich, S,Mobasheri, A(2008).Resveratrol suppresses interleukin-1β-induced inflammatory signaling and apoptosis in human articular chondrocytes: potential for use as a novel nutraceutical for the treatment of osteoarthritis.Biochem Pharmacol,76(11),1426-1439.
30. Shen, CL,Smith, BJ,Lo, DF(2012).Dietary polyphenols and mechanisms of osteoarthritis.J Nutr Biochem,23(11),1367-1377.
31. Shibata, Y,Kume, N,Arai, H(2007).Mulberry leaf aqueous fractions inhibit TNF- α-induced nuclear factor kB (NF-kB) activation and lectin-like oxidized LDL receptor-1 (LOX-1) expression in vascular endothelial cells.Atherosclerosis,193(1),20-27.
32. Soonthornsit, N,Pitaksutheepong, C,Hemstapat, W,Utaisincharoen, P,Pitaksuteepong, T(2017).In vitro anti-inflammatory activity of Morus alba L. stem extract in LPS-stimulated RAW 264.7 cells.Evid Based Complement Alternat Med,2017
33. Srihapon, K(2018).Phitsanulok, Thailand,Naresuan University.
34. Thongsuk, P(2007).Phitsanulok, Thailand,Naresuan University.
35. Yhiraya, C,Pitaksuteepong, T(2012).Mulberry stem (Morus alba L.) extract in lyotropic liquid crystal of tridecyl salicylate/PGE-7 glyceryl cocoate/water system as a skin whitening product.PACCON,2012,301-305.
36. Zhang, Z,Shi, L(2010).Anti-inflammatory and analgesic properties of cis-mulberroside A from Ramulus mori.Fitoterapia,81(3),214-218.
37. Zhishen, J,Mengcheng, T,Jianming, W(1999).The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals.Food Chem,64(4),555-559.