解毒祛瘀滋阴方对MRL/lpr狼疮小鼠CD4~+T细胞MeCP2信号通路的影响
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摘要
目的探讨解毒祛瘀滋阴方治疗系统性红斑狼疮的可能作用机制。方法 30只MRL/lpr狼疮小鼠随机分成中药组、西药组、模型组各10只,10只C57BL/6小鼠作为对照组。中药组予0.5 g/ml浓度的解毒祛瘀滋阴方水煎剂灌胃,西药组予0.5 mg/ml浓度的醋酸泼尼松片混悬液灌胃,模型组和对照组予蒸馏水灌胃,均每次0.01 ml/g,每日1次,连续干预12周。干预结束后观察小鼠皮毛、活动及浅表淋巴结等一般状态,检测小鼠血清白细胞介素4(IL-4)、γ干扰素(IFN-γ)水平变化;HE染色染色观察肾脏病理改变;检测小鼠脾脏CD4~+T细胞甲基结合蛋白2(MeCP2)m RNA和蛋白的表达水平。结果与模型组比较,中药组小鼠肾脏病理损伤明显改善,血清IL-4、IFN-γ浓度水平降低,MeCP2 m RNA和蛋白表达水平明显升高(P<0.05或P<0.01),且中药组MeCP2蛋白及m RNA表达水平升高较西药组明显(P<0.05或P<0.01)。结论解毒祛瘀滋阴方可能是通过激活MeCP2信号通路,进而抑制系统性红斑狼疮的发病。
Objective To explore the potential mechanism of Jiedu Quyu Ziyin Decoction( 解毒祛瘀滋阴 方,JQZD) in treating systemic lupus erythematosus. Methods Thirty MRL/lpr lupus mice were randomized into Chinese medicine( CM) group,Western medicine( WM) group and the model group,with 10 mice in each group,and the other 10 C57 BL/6 mice were set as the control group. CM group was given 0. 5 g/ml of JQZD by gavage,WM group was given 0. 5 mg/ml of prednisone acetate tablets suspension by gavage,while the model group and the control group were given distilled water orally. All interventions were given 0. 01 ml/g once a day,for 12 weeks. After the intervention,General statuses of fur,activity and superficial lymph nodes of mice were observed. Serum levels of IL-4 and IFN-γ were detected. The pathological changes of kidney were observed with HE staining. The expression levels of MeCP2 m RNA and protein in CD4~+T cells in mice spleen were detected. Results Compared with those in the model group,the pathological changes of kidney significantly improved,the serum levels of IL-4 and IFN-γ decreased,and the expressions of MeCP2 m RNA and protein significantly increased in CM group( P < 0. 05 or P <0. 01). The expressions of MeCP2 protein and m RNA in CM group were significantly higher than that in WM group( P < 0. 05 or P < 0. 01). Conclusion JQZD may inhibit systemic lupus erythematosus through the activation of MeCP2 signaling pathway.
Objective To explore the potential mechanism of Jiedu Quyu Ziyin Decoction( 解毒祛瘀滋阴 方,JQZD) in treating systemic lupus erythematosus. Methods Thirty MRL/lpr lupus mice were randomized into Chinese medicine( CM) group,Western medicine( WM) group and the model group,with 10 mice in each group,and the other 10 C57 BL/6 mice were set as the control group. CM group was given 0. 5 g/ml of JQZD by gavage,WM group was given 0. 5 mg/ml of prednisone acetate tablets suspension by gavage,while the model group and the control group were given distilled water orally. All interventions were given 0. 01 ml/g once a day,for 12 weeks. After the intervention,General statuses of fur,activity and superficial lymph nodes of mice were observed. Serum levels of IL-4 and IFN-γ were detected. The pathological changes of kidney were observed with HE staining. The expression levels of MeCP2 m RNA and protein in CD4~+T cells in mice spleen were detected. Results Compared with those in the model group,the pathological changes of kidney significantly improved,the serum levels of IL-4 and IFN-γ decreased,and the expressions of MeCP2 m RNA and protein significantly increased in CM group( P < 0. 05 or P <0. 01). The expressions of MeCP2 protein and m RNA in CM group were significantly higher than that in WM group( P < 0. 05 or P < 0. 01). Conclusion JQZD may inhibit systemic lupus erythematosus through the activation of MeCP2 signaling pathway.
引文
[1]YU C,GERSHWIN ME,CHANG C.Diagnostic criteria for systemic lupus erythematosus:a critical review[J].J Autoimmun,2014,48-49:10-13.doi:10.1016/j.jaut.2014.01.004.
[2]何尚映,林洁,黄鹏,等.他克莫司胶囊联合醋酸泼尼松龙片治疗系统性红斑狼疮的临床研究[J].中国临床药理学杂志,2017,33(15):398-400.
[3]范永升,温成平,李永伟,等.解毒祛瘀滋阴法治疗SLE对撤减激素和保护下丘脑-垂体-肾上腺轴的作用研究[J].浙江中医药大学学报,2006,30(2):199-202.
[4]陈红波,张利棕,寿旗扬,等.解毒祛瘀滋阴方对MRL/lpr狼疮鼠CD4+T细胞DNA甲基化敏感基因表达的影响[J].上海中医药大学学报,2017,31(1):57-62.
[5]SEKIGAWA I,OKADA M,OGASAWARA H,et al.DNA methylation in systemic lupus erythematosus[J].Lupus,2003,12(2):79-85.
[6]BALADA E,ORDIROS J,SERRANOACEDO S,et al.Transcript levels of DNA methyltransferases DNMT1,DNMT3A and DNMT3B in CD4+T cells from patients with systemic lupus erythematosus[J].Immunology,2008,124(3):339-347.
[7]SAITO M,ISHIKAWA F.The m Cp G-binding domain of human MBD3 does not bind to m Cp G but interacts with Nu RD/Mi2 components HDAC1 and MTA2[J].J Biol Chem,2002,277(38):35434-35439.
[8]WEBB R,WREN JD,JEFFRIES M,et al.Variants within MECP2,a key transcriptional regulator,are associated with increased susceptibility to lupus and differential gene expression in lupus patients[J].Arthritis Rheum,2009,60(4):1076-1084.
[9]陈健,范永升,温成平,等.解毒祛瘀滋阴方对狼疮鼠脾淋巴细胞凋亡及相关基因的调控[J].中华中医药杂志,2009,24(12):1560-1563.
[10]BALLESTAR E.Epigenetic alterations in autoimmune rheumatic diseases[J].Nat Rev Rheumatol,2011,7(5):263-271.
[11]SHIOZAWA S.Cause of systemic lupus erythematosus:a novel self-organized criticality theory of autoimmunity[J].Expert Rev Clin Immunol,2014,7(6):715-717.
[12]SUNAHORI K,JUANG YT,KYTTARIS VC,et al.Promoter hypomethylation results in increased expression of protein phosphatase 2A in T cells from patients with systemic lupus erythematosus[J].J Immunol,2011,186(7):4508-4517.
[13]NAN X,CAMPOY FJ,BIRD A.Me CP2 is a transcriptional repressor with abundant binding sites in genomic chromatin[J].Cell,1997,88(4):471-481.
[14]HUGHES T,WEBB R,FEI Y,et al.DNA methylome in human CD4+T cells identifies transcriptionally repressive and no N-repressive methylation peaks[J].Genes Immun,2010,11(7):554-560.
[15]HAI L,YIN H,LING W,et al.The critical role of epigenetics in systemic lupus erythematosus and autoimmunity[J].J Autoimmun,2016,74:118-138.doi:10.1016/j.jaut.2016.06.020.
[16]ZHANG Y,MING Z,SAWALHA AH,et al.Impaired DNA methylation and its mechanisms in CD4+T cells of systemic lupus erythematosus[J].J Autoimmun,2013,41:92-99.doi:10.1016/j.jaut.2013.01.005.
[2]何尚映,林洁,黄鹏,等.他克莫司胶囊联合醋酸泼尼松龙片治疗系统性红斑狼疮的临床研究[J].中国临床药理学杂志,2017,33(15):398-400.
[3]范永升,温成平,李永伟,等.解毒祛瘀滋阴法治疗SLE对撤减激素和保护下丘脑-垂体-肾上腺轴的作用研究[J].浙江中医药大学学报,2006,30(2):199-202.
[4]陈红波,张利棕,寿旗扬,等.解毒祛瘀滋阴方对MRL/lpr狼疮鼠CD4+T细胞DNA甲基化敏感基因表达的影响[J].上海中医药大学学报,2017,31(1):57-62.
[5]SEKIGAWA I,OKADA M,OGASAWARA H,et al.DNA methylation in systemic lupus erythematosus[J].Lupus,2003,12(2):79-85.
[6]BALADA E,ORDIROS J,SERRANOACEDO S,et al.Transcript levels of DNA methyltransferases DNMT1,DNMT3A and DNMT3B in CD4+T cells from patients with systemic lupus erythematosus[J].Immunology,2008,124(3):339-347.
[7]SAITO M,ISHIKAWA F.The m Cp G-binding domain of human MBD3 does not bind to m Cp G but interacts with Nu RD/Mi2 components HDAC1 and MTA2[J].J Biol Chem,2002,277(38):35434-35439.
[8]WEBB R,WREN JD,JEFFRIES M,et al.Variants within MECP2,a key transcriptional regulator,are associated with increased susceptibility to lupus and differential gene expression in lupus patients[J].Arthritis Rheum,2009,60(4):1076-1084.
[9]陈健,范永升,温成平,等.解毒祛瘀滋阴方对狼疮鼠脾淋巴细胞凋亡及相关基因的调控[J].中华中医药杂志,2009,24(12):1560-1563.
[10]BALLESTAR E.Epigenetic alterations in autoimmune rheumatic diseases[J].Nat Rev Rheumatol,2011,7(5):263-271.
[11]SHIOZAWA S.Cause of systemic lupus erythematosus:a novel self-organized criticality theory of autoimmunity[J].Expert Rev Clin Immunol,2014,7(6):715-717.
[12]SUNAHORI K,JUANG YT,KYTTARIS VC,et al.Promoter hypomethylation results in increased expression of protein phosphatase 2A in T cells from patients with systemic lupus erythematosus[J].J Immunol,2011,186(7):4508-4517.
[13]NAN X,CAMPOY FJ,BIRD A.Me CP2 is a transcriptional repressor with abundant binding sites in genomic chromatin[J].Cell,1997,88(4):471-481.
[14]HUGHES T,WEBB R,FEI Y,et al.DNA methylome in human CD4+T cells identifies transcriptionally repressive and no N-repressive methylation peaks[J].Genes Immun,2010,11(7):554-560.
[15]HAI L,YIN H,LING W,et al.The critical role of epigenetics in systemic lupus erythematosus and autoimmunity[J].J Autoimmun,2016,74:118-138.doi:10.1016/j.jaut.2016.06.020.
[16]ZHANG Y,MING Z,SAWALHA AH,et al.Impaired DNA methylation and its mechanisms in CD4+T cells of systemic lupus erythematosus[J].J Autoimmun,2013,41:92-99.doi:10.1016/j.jaut.2013.01.005.