蜂毒肽通过Th1/Th2途径对胶原诱导关节炎大鼠模型免疫调节的影响
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摘要
目的探讨蜂毒肽对胶原诱导关节炎大鼠模型免疫调节的影响。方法成功建立胶原诱导关节炎大鼠模型,随机分为模型组、蜂毒肽(0.5 mg/kg)组、甲氨蝶呤(1 mg/只)组,每组6只,另选取未造模大鼠为对照组;造模第14天起,隔日1次ip蜂毒肽,每周1次ip甲氨蝶呤,对照组、模型组ip等量生理盐水,给药21 d。观察关节肿胀程度、进行关节炎指数(AI)评分;给药结束后,收集血清,ELISA法检测血清中免疫球蛋白G(IgG)、IgG1、IgG2a、干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)水平;分离大鼠脾脏,提取淋巴细胞,流式细胞术检测Th1、Th2细胞比例。结果蜂毒肽组大鼠关节皮肤发红,关节肿胀明显减轻;与模型组比较,蜂毒肽显著抑制AI评分(P<0.05);显著降低血清中IgG、IgG2a、IFN-γ浓度(P<0.05),升高IgG1、IL-4浓度(P<0.05);显著下调Th1细胞比例、Th1/Th2比值(P<0.05),上调Th2细胞比例(P<0.05)。结论蜂毒肽能通过调节Th1/Th2平衡抑制胶原诱导关节炎大鼠的炎症。
Objective To investigate the effect of melittin on the immune regulation of arthritis rat model induced by collagen.Methods The arthritis rat model was successfully established and randomly divided into model group, melittin group and methotrexate group with six rats in each group. In addition, rats without modeling were selected as control group. After 14 d, the melittin were ip injected once every other day and methotrexate were ip given once a week for 21 d. The model group was given the same amount of normal saline, and the degree of joint swelling was observed and calculated. After drug administration, the serum was collected from rats, and the serum levels of IgG, IgG1, IgG2a, IFN-γ, and IL-4 were detected by ELISA. The spleen was isolated from rats and lymphocytes were extracted. The proportion of Th1 and Th2 cells was detected by flow cytometry. Results In the melittin group, there was redness in the joint skin, and the swelling of the joint was obviously alleviated. Compared with model group, melittin significantly inhibited joint swelling score (P < 0.05), reduced the concentrations of IgG, IgG2a, and IFN-γ in serum (P < 0.05), increased concentrations of IgG1 and IL-4 (P < 0.05), down-regulated the ratio of Th1 cells and the ratio of Th1/Th2 (P < 0.05), and up-regulated the ratio of Th2 cells (P < 0.05). Conclusion Melittin can inhibit the inflammation in the collageninduced arthritis by regulating Th1/Th2 balance.
Objective To investigate the effect of melittin on the immune regulation of arthritis rat model induced by collagen.Methods The arthritis rat model was successfully established and randomly divided into model group, melittin group and methotrexate group with six rats in each group. In addition, rats without modeling were selected as control group. After 14 d, the melittin were ip injected once every other day and methotrexate were ip given once a week for 21 d. The model group was given the same amount of normal saline, and the degree of joint swelling was observed and calculated. After drug administration, the serum was collected from rats, and the serum levels of IgG, IgG1, IgG2a, IFN-γ, and IL-4 were detected by ELISA. The spleen was isolated from rats and lymphocytes were extracted. The proportion of Th1 and Th2 cells was detected by flow cytometry. Results In the melittin group, there was redness in the joint skin, and the swelling of the joint was obviously alleviated. Compared with model group, melittin significantly inhibited joint swelling score (P < 0.05), reduced the concentrations of IgG, IgG2a, and IFN-γ in serum (P < 0.05), increased concentrations of IgG1 and IL-4 (P < 0.05), down-regulated the ratio of Th1 cells and the ratio of Th1/Th2 (P < 0.05), and up-regulated the ratio of Th2 cells (P < 0.05). Conclusion Melittin can inhibit the inflammation in the collageninduced arthritis by regulating Th1/Th2 balance.
引文
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[8]Nah S S,Ha E,Mun S H,et al.Effects of melittin on the production of matrix metalloproteinase-1 and-3 in rheumatoid arthritic fibroblast-like synoviocytes[J].JPharmacol Sci,2008,106(1):162-166.
[9]Li J H,Ke T,He C,et al.The anti-arthritic effects of synthetic melittin on the complete Freund's adjuvantinduced rheumatoid arthritis model in rats[J].Am J Chin Med,2010,38(6):1039-1049.
[10]van Roon J A,van Roy J L,Duits A,et al.Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T helper-1 activation is inhibited by interleukin-4[J].Ann Rheum Dis,1995,54(10):836-840.
[11]Lefranc G,Chaabani H,Van Loghem E,et al.Simultaneous absence of the human IgG1,IgG2,IgG4 and IgA1 subclasses:immunological and immunogenetical considerations[J].Eur J Immunol,1983,13(3):240-244.
[12]Somerfield S D,Stach J L,Mraz C,et al.Bee venom melittin blocks neutrophil O2-production[J].Inflammation,1986,10(2):175-182.
[13]Chen L N,Wang C H,Leng N,et al.Combined treatment of etanercept and MTX reverses Th1/Th2,Th17/Treg imbalance in patients with rheumatoid arthritis[J].J Clin Immunol,2011,31(4):596-605.
[2]Manivel V A,Sohrabian A,Wick M C,et al.Anti-type IIcollagen immune complex-induced granulocyte reactivity is associated with joint erosions in RA patients with anticollagen antibodies[J].Arthritis Res Ther,2015,17:8.
[3]Lubberts E.Role of T lymphocytes in the development of rheumatoid arthritis.Implications for treatment[J].Curr Pharm Des,2015,21(2):142-146.
[4]Cameron M,Gagnier J J,Little C V,et al.Evidence of effectiveness of herbal medicinal products in the treatment of arthritis.Part 2:Rheumatoid arthritis[J].Phytother Res,2009,23(12):1647-1662.
[5]Dolhain R J,van der Heiden A N,ter Haar N T,et al.Shift toward T lymphocytes with a T helper 1 cytokinesecretion profile in the joints of patients with rheumatoid arthritis[J].Arthritis Rheum,1996,39(12):1961-1969.
[6]Li J H,Ke T,He C,et al.The anti-arthritic effects of synthetic melittin on the complete Freund's adjuvantinduced rheumatoid arthritis model in rats[J].Am J Chin Med,2010,38(6):1039-1049.
[7]Kim S K,Park K Y,Yoon W C,et al.Melittin enhances apoptosis through suppression of IL-6/sIL-6R complexinduced NF-κB and STAT3 activation and Bcl-2expression for human fibroblast-like synoviocytes in rheumatoid arthritis[J].Joint Bone Spine,2011,78(5):471-477.
[8]Nah S S,Ha E,Mun S H,et al.Effects of melittin on the production of matrix metalloproteinase-1 and-3 in rheumatoid arthritic fibroblast-like synoviocytes[J].JPharmacol Sci,2008,106(1):162-166.
[9]Li J H,Ke T,He C,et al.The anti-arthritic effects of synthetic melittin on the complete Freund's adjuvantinduced rheumatoid arthritis model in rats[J].Am J Chin Med,2010,38(6):1039-1049.
[10]van Roon J A,van Roy J L,Duits A,et al.Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T helper-1 activation is inhibited by interleukin-4[J].Ann Rheum Dis,1995,54(10):836-840.
[11]Lefranc G,Chaabani H,Van Loghem E,et al.Simultaneous absence of the human IgG1,IgG2,IgG4 and IgA1 subclasses:immunological and immunogenetical considerations[J].Eur J Immunol,1983,13(3):240-244.
[12]Somerfield S D,Stach J L,Mraz C,et al.Bee venom melittin blocks neutrophil O2-production[J].Inflammation,1986,10(2):175-182.
[13]Chen L N,Wang C H,Leng N,et al.Combined treatment of etanercept and MTX reverses Th1/Th2,Th17/Treg imbalance in patients with rheumatoid arthritis[J].J Clin Immunol,2011,31(4):596-605.