拟黑多刺蚁活性组分对系统性红斑狼疮的治疗作用及机制
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摘要
目的:采用动物模型和细胞模型,探讨拟黑多刺蚁活性组分(AFPV)对系统性红斑狼疮(SLE)的作用及可能机制。方法:采用大鼠Arthus反应,将60只SD大鼠随机分为正常组,模型组,醋酸泼尼松组(5 mg·kg~(-1)),AFPV高、中、低剂量组(400,200,100 mg·kg~(-1))。采用牛血清蛋白-弗氏完全(不完全)佐剂复制SLE模型。观察AFPV对SLE大鼠背部皮肤红肿直径的影响,酶联免疫吸附测定检测血清抗双链DNA(ds DNA)抗体,补体3(C3),补体4(C4),免疫球蛋白M(Ig M),白细胞介素-1(IL-1),白细胞介素-6(IL-6),白细胞介素-31(IL-31),白细胞介素-33(IL-33)水平;免疫磁珠法分选16～18周MRL/lpr狼疮小鼠和C57BL/6J正常小鼠脾脏CD4+T细胞,观察AFPV对miR-200a,miR-155的表达及沉默E盒锌指结合蛋白1(ZEB1)和细胞因子信号转导抑制因子1(SOCS1)水平的影响,探讨AFPV对SLE的作用及可能机制。结果:与正常组比较,模型组大鼠背部皮肤红肿直径显著增大(P <0. 01),与模型组比较,AFPV高、中剂量组可显著降低SLE大鼠背部皮肤红肿直径(P <0. 05,P <0. 01);与正常组比较,模型组大鼠血清Ig M,IL-6,IL-33明显增高(P <0. 05),与模型组大鼠比较,AFPV高、中给药组Ig M,IL-6,IL-33明显降低(P <0. 05);与正常组比较,MRL/lpr狼疮小鼠的CD4+T细胞miR-200a的表达显著降低,miR-155的表达显著增高及ZEB1水平显著增高而SOCS1的表达显著降低(P <0. 01),与模型组比较,AFPV干预后miR-200a的表达显著增高,miR-155的表达显著降低(P <0. 01),ZEB1水平显著降低而SOCS1的表达显著增高(P <0. 01)。结论:AFPV对SLE大鼠具有一定的治疗作用,其机制可能与其对miR-200a,miR-155及ZEB1和SOCS1的调节有关。
Objective: To observe the effect of an active fraction of Polyrhachis vicina( AFPV) on systemic lupus erythematosus( SLE) and its possible mechanism based on animal and cell models. Method:Totally 60 SD rats were randomly divided into normal control group,model group,prednisone acetate group( 5 mg·kg~(-1)),and high,medium and low-dose AFPV groups( 400,200,100 mg·kg~(-1)). SLE model was replicated with bovine serum albumin-Freund's complete( incomplete) adjuvant. Arthus reaction was observed to study the effect of AFPV on the diameter of back skin redness in rats with SLE. The expressions of anti-doublestranded DNA( dsDNA) antibody,complements 3( C3),complement 4( C4),immunoglobulin M( Ig M),interleukin~(-1)( IL-1),interleukin-6( IL-6),interleukin-31( IL-31) and interleukin-33( IL-33) in serum were detected by enzyme-linked immunosorbent assay. CD4+T cells were isolated from the spleens of MRL/lpr and C57 BL/6 J mice at the age of 16 to 18 weeks by immunomagnetic beads method. The expressions of miR-200 a and miR-155 and the levels of zinc-finger-enhancer binding protein 1( ZEB1) and suppressor of cytokine signaling1( SOCS1) in CD4+T cells were observed to explore the effect of AFPV on SLE and its possible mechanism.Result: Compared with the normal group,the diameter of back skin swelling in the model group was significantly increased( P < 0. 01). Compared with the model group,the high and medium-dose AFPV groups significantly reduce the skin redness on the back of SLE rats( P < 0. 05,P < 0. 01); Compared with the normal group,the serum levels of Ig M,IL-6 and IL-33 were significantly increased in the model group( P < 0. 05). Compared with the model group,the serum levels of IgM,IL-6 and IL-33 were significantly decreased after the intervention of AFPV( P < 0. 05). Compared with the normal group,the expression of miR-200 a was significantly decreased( P <0. 01),and its target protein ZEB1 was significantly increased( P < 0. 01) in the CD4+T cells of MRL/lpr lupus mice. Compared with the model group,the expression of microRNA-200 a increased significantly,the expression of microRNA-155 decreased significantly( P < 0. 01),the level of ZEB1 decreased significantly,and the expression of SOCS1 increased significantly after AFPV intervention( P < 0. 01). Conclusion: AFPV has therapeutic effect on rats with SLE,its mechanism may be related to the regulation of miR-200 a/ZEB1 and miR-155/SOCS1.
Objective: To observe the effect of an active fraction of Polyrhachis vicina( AFPV) on systemic lupus erythematosus( SLE) and its possible mechanism based on animal and cell models. Method:Totally 60 SD rats were randomly divided into normal control group,model group,prednisone acetate group( 5 mg·kg~(-1)),and high,medium and low-dose AFPV groups( 400,200,100 mg·kg~(-1)). SLE model was replicated with bovine serum albumin-Freund's complete( incomplete) adjuvant. Arthus reaction was observed to study the effect of AFPV on the diameter of back skin redness in rats with SLE. The expressions of anti-doublestranded DNA( dsDNA) antibody,complements 3( C3),complement 4( C4),immunoglobulin M( Ig M),interleukin~(-1)( IL-1),interleukin-6( IL-6),interleukin-31( IL-31) and interleukin-33( IL-33) in serum were detected by enzyme-linked immunosorbent assay. CD4+T cells were isolated from the spleens of MRL/lpr and C57 BL/6 J mice at the age of 16 to 18 weeks by immunomagnetic beads method. The expressions of miR-200 a and miR-155 and the levels of zinc-finger-enhancer binding protein 1( ZEB1) and suppressor of cytokine signaling1( SOCS1) in CD4+T cells were observed to explore the effect of AFPV on SLE and its possible mechanism.Result: Compared with the normal group,the diameter of back skin swelling in the model group was significantly increased( P < 0. 01). Compared with the model group,the high and medium-dose AFPV groups significantly reduce the skin redness on the back of SLE rats( P < 0. 05,P < 0. 01); Compared with the normal group,the serum levels of Ig M,IL-6 and IL-33 were significantly increased in the model group( P < 0. 05). Compared with the model group,the serum levels of IgM,IL-6 and IL-33 were significantly decreased after the intervention of AFPV( P < 0. 05). Compared with the normal group,the expression of miR-200 a was significantly decreased( P <0. 01),and its target protein ZEB1 was significantly increased( P < 0. 01) in the CD4+T cells of MRL/lpr lupus mice. Compared with the model group,the expression of microRNA-200 a increased significantly,the expression of microRNA-155 decreased significantly( P < 0. 01),the level of ZEB1 decreased significantly,and the expression of SOCS1 increased significantly after AFPV intervention( P < 0. 01). Conclusion: AFPV has therapeutic effect on rats with SLE,its mechanism may be related to the regulation of miR-200 a/ZEB1 and miR-155/SOCS1.
引文
[1] Gergianaki I,Bortoluzzi A,Bertsias G. Update on the epidemiology,risk factors,and disease outcomes of systemic lupus erythematosus[J]. Best Pract Res Clin Rheumatol,2018,32(2):188-205.
[2] El Qadiry R,Bourrahouat A,Aitsab I,et al. Systemic lupus erythematosus-related pancreatitis in children:severe and lethal form[J]. Case Rep Pediatr,2018,9:e4612754.
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[9]王璟,汪悦,夏卫军,等.解毒祛斑汤对大鼠Arthus反应的影响[J].中国中医急症,2007,16(2):204-205.
[10]张萍,刘红云,刘晓燕,等.调节性T细胞在小鼠ITP发病中的作用[J].中国实验血液学杂志,2016,24(3):784-787.
[11]罗建蓉,肖怀,张成桂,等.两种药用蜚蠊油脂成分的GC-MS分析[J].中国实验方剂学杂志,2017,23(11):55-58.
[12]杜晨晖,闫艳,朱羽尧,等.山西不同产地酸枣仁中脂肪酸类成分的GC-MS分析[J].中国实验方剂学杂志,2018,24(24):19-25.
[13] SU Q B,SU H,NONG Z H,et al. Hypouricemic and nephroprotective effects of an active fraction from Polyrhachis vicina Roger on potassium oxonate-induced hyperuricemia in rats[J]. Kidney Blood Press Res,2018,43(1):220-233.
[14]曹文婷,孙萍,李华艳,等. n-3多不饱和脂肪酸在不同疾病中的研究进展[J].山西医药杂志,2014,43(14):1668-1670.
[15]徐丽敏,陈学荣,屠呦呦.双氢青蒿素对红斑狼疮性BXSB小鼠的作用[J].中国中西医结合皮肤性病学杂志,2002,1(1):19-20.
[16] MO Y M,FANG P H,WAI K I,et al. Serum levels of IL-33 and soluble ST2 and their association with disease activity in systemic lupus erythematosus[J].Rheumatology(Oxford),2010,49(3):520-527.
[17] Düster M,Becker M,Gnirck A C,et al. T cell-derived IFN-downregulates protective group 2 innate lymphoid cells in murine lupus erythematosus[J]. Eur J Immunol,2018,48(8):1364-1375.
[18] GUO J,XIANG Y,PENG Y F,et al. The association of novel IL-33 polymorphisms with s IL-33 and risk of systemic lupus erythematosus[J]. Mol Immunol,2016,77:1-7.
[19] WAN A W,Mohd Shahrir M S,Asrul W,et al. The association between serum prolactin levels and interleukin-6 and systemic lupus erythematosus activity[J]. Reumatismo,2018,70(4):241-250.
[20] Karampetsou M P,Comte D,Suárez-Fueyo A,et al.Signaling lymphocytic activation molecule family member1 engagement inhibits T cell-B cell interaction and diminishes interleukin-6 production and plasmablast differentiation in systemic lupus erythematosus[J].Arthritis Rheumatol,2019,71(1):99-108.
[21] Lashine Y A,Salah S,Aboelenein H R,et al. Correcting the expression f miRNA-155 represses PP2AC and enhances the release of IL-2 in pbmcs of juvenile SLE patients[J]. Lupus,2015,24(3):240-247.
[2] El Qadiry R,Bourrahouat A,Aitsab I,et al. Systemic lupus erythematosus-related pancreatitis in children:severe and lethal form[J]. Case Rep Pediatr,2018,9:e4612754.
[3] Ali A,Sayyed Z,Ameer M A,et al. Systemic lupus erythematosus:an overview of the disease pathology and its management[J]. Cureus,2018,10(9):e3288.
[4]何飞,李冬梅,苏启表,等.拟黑多刺蚁活性组分治疗小鼠皮肤瘙痒的实验研究[J].中药材,2018,41(5):1201-1204.
[5]韦桂宁,曾宪彪,牙启康,等.拟黑多翅蚁抗痛风作用研究[J].中国实验方剂学杂志,2012,18(20):205-208.
[6]张欣,龙倩,楚世峰,等.拟黑多刺蚁石油醚部位对抑郁大鼠神经炎症反应的抑制作用[J].药学学报,2018,53(7):1042-1047.
[7] SU Q,SU H,NONG Z,et al. Hypouricemic and nephroprotective effects of an active fraction from polyrhachis vicina roger on potassium oxonate-induced hyperuricemia in rats[J]. Kidney Blood Press Res,2018,43(1):220-233.
[8] WEI G N,CHU S F,SU Q B,et al. Antidepressant-like effect of active fraction of Polyrhachisvicina Roger in a rat depression model[J]. J Tradit Chin Med,2018,38(1):12-21.
[9]王璟,汪悦,夏卫军,等.解毒祛斑汤对大鼠Arthus反应的影响[J].中国中医急症,2007,16(2):204-205.
[10]张萍,刘红云,刘晓燕,等.调节性T细胞在小鼠ITP发病中的作用[J].中国实验血液学杂志,2016,24(3):784-787.
[11]罗建蓉,肖怀,张成桂,等.两种药用蜚蠊油脂成分的GC-MS分析[J].中国实验方剂学杂志,2017,23(11):55-58.
[12]杜晨晖,闫艳,朱羽尧,等.山西不同产地酸枣仁中脂肪酸类成分的GC-MS分析[J].中国实验方剂学杂志,2018,24(24):19-25.
[13] SU Q B,SU H,NONG Z H,et al. Hypouricemic and nephroprotective effects of an active fraction from Polyrhachis vicina Roger on potassium oxonate-induced hyperuricemia in rats[J]. Kidney Blood Press Res,2018,43(1):220-233.
[14]曹文婷,孙萍,李华艳,等. n-3多不饱和脂肪酸在不同疾病中的研究进展[J].山西医药杂志,2014,43(14):1668-1670.
[15]徐丽敏,陈学荣,屠呦呦.双氢青蒿素对红斑狼疮性BXSB小鼠的作用[J].中国中西医结合皮肤性病学杂志,2002,1(1):19-20.
[16] MO Y M,FANG P H,WAI K I,et al. Serum levels of IL-33 and soluble ST2 and their association with disease activity in systemic lupus erythematosus[J].Rheumatology(Oxford),2010,49(3):520-527.
[17] Düster M,Becker M,Gnirck A C,et al. T cell-derived IFN-downregulates protective group 2 innate lymphoid cells in murine lupus erythematosus[J]. Eur J Immunol,2018,48(8):1364-1375.
[18] GUO J,XIANG Y,PENG Y F,et al. The association of novel IL-33 polymorphisms with s IL-33 and risk of systemic lupus erythematosus[J]. Mol Immunol,2016,77:1-7.
[19] WAN A W,Mohd Shahrir M S,Asrul W,et al. The association between serum prolactin levels and interleukin-6 and systemic lupus erythematosus activity[J]. Reumatismo,2018,70(4):241-250.
[20] Karampetsou M P,Comte D,Suárez-Fueyo A,et al.Signaling lymphocytic activation molecule family member1 engagement inhibits T cell-B cell interaction and diminishes interleukin-6 production and plasmablast differentiation in systemic lupus erythematosus[J].Arthritis Rheumatol,2019,71(1):99-108.
[21] Lashine Y A,Salah S,Aboelenein H R,et al. Correcting the expression f miRNA-155 represses PP2AC and enhances the release of IL-2 in pbmcs of juvenile SLE patients[J]. Lupus,2015,24(3):240-247.