姜黄素对乳腺炎模型小鼠细胞免疫的影响
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摘要
探讨姜黄素(curcumin)对致病性金黄色葡萄球菌(Staphylococcus aureus,S.aureus)所诱导的乳腺炎模型小鼠血液中T淋巴细胞亚群及脾脏中NK细胞活性的影响。构建小鼠乳腺炎模型,分别通过流式细胞术、ELISA法、MTT法和平板计数法检测空白对照组、模型组、阳性对照组、姜黄素组及姜黄乳炎散组血液中CD4~+T、CD8~+T活性和血清中IL-1、IL-2、IL-10的水平,脾脏中NK细胞活性及S.aureus数量的变化差异来监测各组药物抗炎效果。结果显示,相比于模型组,姜黄素组、姜黄乳炎散组和阳性对照组可显著增加乳腺炎小鼠血清中IL-2、IL-10的水平以及脾脏中NK细胞和血液中CD4~+T的活性,显著降低小鼠血清中IL-1的水平以及血液中CD8~+T活性(P<0.05),以姜黄素组效果最佳。结果表明,姜黄素对小鼠乳腺炎具有保护作用,并且姜黄素对S.aureus所诱导的小鼠乳腺炎的抗炎机制可能通过抑制IL-1和CD8~+T水平,增强IL-10、NK、CD4~+T和减少S.aureus的数量而发挥作用。姜黄素可能成为潜在的乳房炎治疗药物。
The aim of this study was to investigate the effects of curcumin on the T lymphocyte subsets in blood and NK cell activity in the spleen of mastitis model mice induced by pathogenic Staphylococcus aureus(S.aureus).The changes of CD4~+T,CD8~+T in blood,IL-1,IL-2 and IL-10 levels in serum,NK cell activity in spleen and S.aureus quantity in blank control group,model group,positive control group,curcumin group and Terunshudule(TRSDL) group were detected by flow cytometry,ELISA,MTT and plate counting methods respectively to monitor the anti-inflammatory effect of each group.The curcumin group,TRSDL group and positive control group increased the levels of IL-2 and IL-10 in serum,activity of spleen NK cells and blood CD4~+T of mastitis mice,and reduced the levels of IL-1 in serum and CD8~+T activity in blood of mice compared with the model group(P≤0.05),of wich curcumin group was the best effect.This study shows that curcumin has a protective effect on mastitis mouse,and the anti-inflammatory effect of curcumin on S.aureus induced mastitis mouse may be inhibit IL-1 and CD8~+T levels,enhance IL-10,NK,CD4~+T and reduce the number of S.aureus.curcumin may become a potential treatment drug for mastitis.
The aim of this study was to investigate the effects of curcumin on the T lymphocyte subsets in blood and NK cell activity in the spleen of mastitis model mice induced by pathogenic Staphylococcus aureus(S.aureus).The changes of CD4~+T,CD8~+T in blood,IL-1,IL-2 and IL-10 levels in serum,NK cell activity in spleen and S.aureus quantity in blank control group,model group,positive control group,curcumin group and Terunshudule(TRSDL) group were detected by flow cytometry,ELISA,MTT and plate counting methods respectively to monitor the anti-inflammatory effect of each group.The curcumin group,TRSDL group and positive control group increased the levels of IL-2 and IL-10 in serum,activity of spleen NK cells and blood CD4~+T of mastitis mice,and reduced the levels of IL-1 in serum and CD8~+T activity in blood of mice compared with the model group(P≤0.05),of wich curcumin group was the best effect.This study shows that curcumin has a protective effect on mastitis mouse,and the anti-inflammatory effect of curcumin on S.aureus induced mastitis mouse may be inhibit IL-1 and CD8~+T levels,enhance IL-10,NK,CD4~+T and reduce the number of S.aureus.curcumin may become a potential treatment drug for mastitis.
引文
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[2] BURVENICH C,VAN MERRTS V,MEHRZAD J,et al.Severity of E.coli mastitis is mainly determined by cow factors[J].Vet Res,2003,34(5):521-564.
[3] GRUET P,MAINCENT P,BERTHELOT X,et al.Bovine mastitis and intramammary drug delivery:review and perspectives[J].Adv Drug Deliv Rev,2001,50:245-259.
[4] LIU Q Z,WU Q,ZHANG Y B,et al.Prevalence of clinical meticillin-resistant Staphylococcus aureus(MRSA) with high-level mupirocin resistance in Shanghai and Wenzhou,China[J].Int J Antimicrobial Agents,2010,35(2):114-118.
[5] GUNES H,GULEN D,MUTLU R,et al.Antibacterial effects of curcumin:an in vitro minimum inhibitory concentration study[J].Toxicol Ind Health,2016,32(2):246.
[6] LU Y C,YEH W C,OHASHI P S.LPS/TLR4 signal transduction pathway[J].Cytokine,2008,42(2):145-151.
[7] CHUSRI S,JITTANON W,MANEENOON K,et al.An effective antibio?lm agent against Pseudomonas aeruginosa bio?lm from traditional Thai herbal recipes used for wound treatments[J].Microbial Drug Resist,2013,19:337-343.
[8] CHUSRI S,SOMPETCH K,MUKDEE S,et al.Inhibition of Staphylococcus epidermidis bio?lm formation by traditional Thai herbal recipes used for wound treatment[J].Evid Based Complement Alternat Med,2012,2012:159797.
[9] KANEKO K,AOKI H,FURUICHI T,et al.Influence of uterine inflammation on the estrous cycle in rats[J].J Reprod Dev,2004,50(3):361-367.
[10] AQQARWAL B B,HARIKUMAR K B.Potential therapeutic effect of curcumin,the anti-inflammatory agent,against neurodegenerative,cardiovascular,pulmonary,metabolic autoimmune and neoplastic diseases[J].Int J Biochem Cell Biol,2009,41(1):40.
[11] YUN S S,KIM S P,KANG M Y,et al.Inhibitory effect of curcumin on liver sepsis-injury in a murine model of endotoxemic shock[J].Biotechnol Lett,2010,32:209-214.
[12] 战永波.治疗奶牛乳房炎蒙药复方的筛选及其抗炎免疫机理的研究[D].内蒙古呼和浩特:内蒙古农业大学,2010.
[13] FU Y H,GAO R F,CAO M Y,et al.Curcumin attenuates inflammatory responses by suppressing TLR4-mediated NF-κB signaling pathway in lipopolysaccharide-induced mastitis in mice[J].Int Immunopharmacol,2014,20(1):54-58.
[14] 贾知锋,王纯洁,敖日格乐,等.Nisin及大黄复方对子宫内膜炎大鼠血清中MMP-2和MMP-9活性的影响[J].中国农业大学学报,2017(6):109-115.
[15] 徐进,敖日格乐,贾知锋,等.姜黄素对乳腺炎模型小鼠血清中抗炎因子的影响[J].中国农业大学学报,2018,23(4):89-74.
[16] 王斌,李京京,李秋荣,等.乳杆菌对致病性大肠杆菌感染小鼠肠黏膜屏障功能的影响[J].肠外与肠内营养,2007,14(6):321-325.
[17] 郭婧.治疗奶牛乳房炎的抗炎免疫蒙兽药成分和成分复方的筛选及其作用机理研究[D].内蒙古呼和浩特:内蒙古农业大学,2014.
[18] ZHAO H M,HAN F,XU R,et al.Therapeutic effect of curcumin on experimental colitis mediated by inhibiting CD8 CD11c cells[J].World J Gastroenterol,2017,23(10):1804-1815.
[19] HOGEVEEN H,HUIJPS K,LAM T J.Economic aspects of mastitis:new developments[J].New Zealand Vet J,2011,59:16-23.
[20] LI D P,FU Y H,ZHANG W,et al.Salidroside attenuates inflammatory responses by suppressing nuclear factor-kappa B and mitogen activated protein kinases activation in lipopolysaccharide-induced mastitis in mice[J].Inflamm Res,2013,62(1):9-15.
[21] WANG W,LIANG D J,SONG X J,et al.Magnolol inhibits the inflammatory response in mouse mammary epithelial cells and a mouse mastitis model[J].Inflammation,2015,38(1):16-26.
[22] FU Y,ZHOU E,LIU Z,et al.Staphylococcus aureus and Escherichia coli elicit different innate immune responses from bovine mammary epithelial cells[J].Vet Immunol Immunopathol,2013,155(4):245-252.
[23] WOLFS T G,BUURMAN W A,VAN SCHADEWIJK A,et al.In vivo expression of Toll-like receptor 2 and 4 by renal epithelial cells:IFN-gamma and TNF-alpha up regulation during inflammation[J].J Immunol,2002,168(3):1286-1293.
[24] KAPLANSKI G,MARIN V,MONTERO-JULIAN F,et al.IL-6:a regulator of the transition from neutrophil to monocyte recruitment during inflammation[J].New J Chem,2003,27(1):80-87.
[25] LI X,YEH V,MOLTENI V.Liver X receptor modulators:a review of recently patented compounds(2007-2009)[J].Expert Opin Ther Pat,2010,20(4):535-562.
[26] SPISNI R,FAILLI A,ORSINI G,et al.Colorectal cancer:tissutal explantation and primary cell culture[J].Ann Ital Chir,2009,80(3):211-217.
[27] RAULET D H.Interplay of natural killer cells and their receptors with the adaptive immune response[J].Nat Immunol,2004,5(10):996-1002.
[28] PETITDEMANGE C,WAUQUIER N,REY J,et al.Control of acute dengue virus infection by natural killer cells[J].Front Immunol,2014,5(5):209.
[29] HONG D S,ANGELO L S,KURZROCK R.Interleukin-6 and its receptor in cancer:implications for translational therapeutics[J].Cancer,2007,110(9):1911.
[30] FUJIWARA D,CHEN L,WEI B,et al.Small intestine CD11c+ CD8+T cells suppress CD4+T cell-induced immune colitis[J].Am J Physiol Gastrointest Liver Physiol,2011,300(6):G939-G947.
[31] LANGLOIS R A,VARBLE A,CHUA M A,et al.Hematopoietic-specific targeting of influenza A virus reveals replication requirements for induction of antiviral immune responses[J].Proc Natl Acad Sci USA,2012,109(30):12117-12122.
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[33] BALASUBRAMANIAN S,ECKERT R L.Keratinocyte proliferation,differentiation,and apoptosis differential mechanisms of regulation by curcumin,EGCG and apigenin[J].Toxicol Appl Pharmacol,2007,224(3):214-219.