乳铁蛋白对IL-1β诱导人骨关节炎滑膜成纤维细胞MMPs、COX-2、PGE2产生的影响
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摘要
目的 探讨乳铁蛋白(LF)对白介素-1β(IL-1β)诱导人骨关节炎(OA)滑膜成纤维细胞中基质金属蛋白酶(MMPs)、环氧化酶-2(COX-2)、前列腺素E2(PGE2)产生的影响。方法 分离并培养人OA滑膜成纤维细胞,取3～4代的细胞进行实验,应用细胞免疫荧光技术来鉴定细胞性质,采用CCK8法检测不同浓度乳铁蛋白对OA滑膜成纤维细胞活性的影响,用实时定量PCR(RT-PCR)和Western blot法检测乳铁蛋白对IL-1β诱导人OA滑膜成纤维细胞中MMPs和COX-2表达的影响,用ELISA法测量PGE2表达的影响。结果 CCK8结果显示,在一定浓度范围内LF对滑膜成纤维细胞活性无明显影响,差异无统计学意义;RT-PCR、Western blot和ELISA结果提示LF对IL-1β诱导人OA滑膜成纤维细胞中MMPs、COX-2、PGE2产生具有抑制作用。结论 LF能够抑制IL-1β诱导人OA滑膜成纤维细胞中MMPs、COX-2、PGE2的表达,对OA具有保护作用。
Objective To investigate the effects of lactoferrin( LF) on interleukin-1β( IL-1β)-induced osteoarthritis( OA) synovial fibroblast matrix metalloproteinases( MMPs) and cyclooxygenase-2( COX-2),prostaglandin E2( PGE2). Methods Human OA synovial fibroblasts were isolated and cultured,and cells of 3 to 4 passages were used for experiments,and cell immunofluorescence technique was used to identify the cell properties. CCK8 method was used to detect different concentrations of lactoferrin on OA synovial fibroblasts. The effects of lactoferrin on the expression of MMPs and COX-2 in human OA synovial fibroblasts induced by IL-1β were detected by real-time quantitative PCR( RT-PCR) and Western blot. The effect of PGE2 expression was measured by ELISA. Results The results of CCK8 showed that LF had no significant effect on synovial fibroblast activity within a certain concentration range,and the difference was not statistically significant. The results of RT-PCR,Western blot and ELISA indicated that LF inhibited the production of MMPs,COX-2 and PGE2 in human OA synovial fibroblasts induced by IL-1β. Conclusion LF can inhibit IL-1β-induced human OA synovial fiber formation. The expression of MMPs,COX-2 and PGE2 in cells has a protective effect on OA.
Objective To investigate the effects of lactoferrin( LF) on interleukin-1β( IL-1β)-induced osteoarthritis( OA) synovial fibroblast matrix metalloproteinases( MMPs) and cyclooxygenase-2( COX-2),prostaglandin E2( PGE2). Methods Human OA synovial fibroblasts were isolated and cultured,and cells of 3 to 4 passages were used for experiments,and cell immunofluorescence technique was used to identify the cell properties. CCK8 method was used to detect different concentrations of lactoferrin on OA synovial fibroblasts. The effects of lactoferrin on the expression of MMPs and COX-2 in human OA synovial fibroblasts induced by IL-1β were detected by real-time quantitative PCR( RT-PCR) and Western blot. The effect of PGE2 expression was measured by ELISA. Results The results of CCK8 showed that LF had no significant effect on synovial fibroblast activity within a certain concentration range,and the difference was not statistically significant. The results of RT-PCR,Western blot and ELISA indicated that LF inhibited the production of MMPs,COX-2 and PGE2 in human OA synovial fibroblasts induced by IL-1β. Conclusion LF can inhibit IL-1β-induced human OA synovial fiber formation. The expression of MMPs,COX-2 and PGE2 in cells has a protective effect on OA.
引文
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[14] Rasheed N,Alghasham A,Rasheed Z,et al.Lactoferrin from camelus dromedarius inhibits nuclear transcription factor-kappa B activation,cyclooxygenase-2 expression and prostaglandin E2 production in stimulated human chondrocytes[J].Pharmacognosy Res,2016,8(2):135-41.
[2] Chang J,Liao Z,Lu M,et al.Systemic and local adipose tissue in knee osteoarthritis[J].Osteoarthritis Cartilage,2018,26(7):864-71.
[3] Moasheri A,Raymann M P,Gualillo O,et al.The role of metabolism in the pathogenesis of osteoarthritis[J].Nat Rev Rheumatol,2017,13(5):302-11.
[4] Rollín R,Marco F,Jover J A,et al.Early lymphocyte activation in the synovial microenvironment in patients with osteoarthritis:comparison with rheumatoid arthritis patients and healthy controls[J].Rheumatol Int,2008,28(8):757-64.
[5] Kitanaka T,Nakano R,Kitanaka N,et al.JNK activation is essential for activation of MEK/ERK signaling in IL-1β-induced COX-2 expression in synovial fibroblasts[J].Sci Rep,2017,7:39914.
[6] Fu Y,Lei J,Zhuang Y,et al.Overexpression of HMGB1 A-box reduced IL-1β-induced MMP expression and the production of inflammatory mediators in human chondrocytes[J].Exp Cell Res,2016,349(1):184-90.
[7] Hao L,Shan Q,Wei J,et al.Lactoferrin:major physiological functions and applications[J].Curr Protein Pept Sci,2019,20(2):139-44.
[8] 肖楚吟,潘云峰,郭兴华,等.滑膜成纤维细胞的原代培养及生物特性[J].中国医学工程,2010,18(2):1-4.
[9] MacFarlane L A,Yang H,Collins J E,et al.Relationship between patient-reported swelling and MRI-defined effusion-synovitis in patients with meniscus tears and knee osteoarthritis[J].Arthritis Care Res(Hoboken),2019,71(3):385-9.
[10] Van Doren S R.Matrix metalloproteinase interactions with collagen and elastin[J].Matrix Biol,2015,44-46:224-31.
[11] Kawahara K,Hohjoh H,Inazumi T,et al.Prostaglandin E2-induced inflammation:relevance of prostaglandin E receptors[J].Biochim Biophys Acta,2015,1851(4):414-21.
[12] Togawa J,Nagase H,Tanaka K,et al.Oral administration of lactoferrin reduces colitis in rats via modulation of the immune system and correction of cytokine imbalance[J].J Gastroenterol Hepatol,2002,17(12):1291-8.
[13] Zeng G Q,Chen A B,Li W,et al.High MMP-1,MMP-2,and MMP-9 protein levels in osteoarthritis[J].Genet Mol Res,2015,14(4):14811-22.
[14] Rasheed N,Alghasham A,Rasheed Z,et al.Lactoferrin from camelus dromedarius inhibits nuclear transcription factor-kappa B activation,cyclooxygenase-2 expression and prostaglandin E2 production in stimulated human chondrocytes[J].Pharmacognosy Res,2016,8(2):135-41.