加味过敏煎对荨麻疹小鼠皮肤肥大细胞脱颗粒的影响
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摘要
目的探讨加味过敏煎治疗荨麻疹的可能作用机制。方法 60只BALB/c小鼠随机分为空白组、单纯IgE组、模型组、过敏煎组、加味过敏煎组、氯雷他定组,每组10只。过敏煎组给予过敏煎10. 27 g/(kg·d)灌胃,加味过敏煎组给予加味过敏煎14. 39 g/(kg·d)灌胃,氯雷他定组给予氯雷他定片2. 10 mg/(kg·d)灌胃,模型组、空白组及单纯IgE组给予等体积生理盐水灌胃,每日1次,连续7次。第6日给药后除空白组外其余各组予尾静脉注射抗DNP IgE单克隆抗体0. 5 ml,次日除空白组和单纯IEg组外其余各组小鼠耳部涂抹2,4-二硝基氟苯50μl激发建立荨麻疹动物模型。第8日检测各组小鼠皮肤肥大细胞脱颗粒情况及蛋白酶激活受体2 (PAR-2) mRNA表达和血清组胺浓度。结果显微镜下观察,空白组与单纯Ig E组未见明显肥大细胞脱颗粒,模型组肥大细胞脱颗粒明显,各给药组肥大细胞脱颗粒数均较模型组低,加味过敏煎组脱颗粒数相对较少。与空白组、单纯IgE组比较,模型组肥大细胞脱颗粒数、PAR-2 mRNA及组胺表达明显增加(P <0. 05或P <0. 01);与模型组比较,加味过敏煎组肥大细胞脱颗粒数、PAR-2 mRNA及组胺表达均明显降低(P <0. 01),且均低于过敏煎组与氯雷他定组(P <0. 05或P <0. 01)。结论加味过敏煎可能通过下调皮肤PAR-2 mRNA表达,降低血清组胺释放,进而抑制皮肤肥大细胞脱颗粒,稳定肥大细胞,以达到防治荨麻疹的作用。
Objective To study the possible mechanism of Modified Guominjian Decoction( 加味过敏煎) in treating urticaria. Methods A total of 60 BALB/c mice were randomly divided into blank group,simple IgE group,model group,Guominjian Decoction group,Modified Guominjian Decoction group and loratadine group,with 10 rats in each group. The Guominjian Decoction group was given Guominjian Decoction 10. 27 g/( kg·d),and the Modified Guominjian Decoction group was given Modified Guominjian Decoction 14. 39 g/( kg · d),the loratadine group was given loratadine pills 2. 10 mg/( kg·d),and the model group,blank group and simple IgE group were given equal volume normal saline,once a day,for constant 7 days. On the 6 th day,except for the blank group,the other groups were injected with 0. 5 ml of anti-DNP IgE monoclonal antibody in the tail vein. The next day,except for the blank group and the simple IgE group,the ears of the other groups were smeared with 2 4-dinitrofluorobenzene 50 μl to activate for the establishment of the animal model of urticaria. On the 8 th day,the degranulation of skin mast cells,the expression of protease activated receptor 2( PAR-2) mRNA and serum histamine concentration were detected in each group. Results With the observation of microscope,there was no obvious degranulation of mast cells in the blank group and the simple IgE group. The degranulation of mast cells in the model group was obvious. The number of degranulation of mast cells in each administration group was lower than that in the model group. The number of degranulation in the Guominjian Decoction group was relatively small. Compared with the blank group and the simple Ig E group,the number of degranulation,PAR-2 mRNA and histamine expression in the model group were significantly increased( P < 0. 05 or P < 0. 01). Compared with the model group,the number of degranulation,PAR-2 mRNA and histamine expression in the Modified Guominjian Decoction group were significantly decreased( P < 0. 01),and were decreased more than those in the Guominjian Decoction group and the loratadine group( P < 0. 05 or P < 0. 01).Conclusion The Modified Guominjian Decoction may down-regulate the expression of PAR-2 in the skin,reduce the release of serum histamine,inhibit the degranulation of skin mast cells,stabilize mast cells to achieve the function of treating and preventing urticaria.
Objective To study the possible mechanism of Modified Guominjian Decoction( 加味过敏煎) in treating urticaria. Methods A total of 60 BALB/c mice were randomly divided into blank group,simple IgE group,model group,Guominjian Decoction group,Modified Guominjian Decoction group and loratadine group,with 10 rats in each group. The Guominjian Decoction group was given Guominjian Decoction 10. 27 g/( kg·d),and the Modified Guominjian Decoction group was given Modified Guominjian Decoction 14. 39 g/( kg · d),the loratadine group was given loratadine pills 2. 10 mg/( kg·d),and the model group,blank group and simple IgE group were given equal volume normal saline,once a day,for constant 7 days. On the 6 th day,except for the blank group,the other groups were injected with 0. 5 ml of anti-DNP IgE monoclonal antibody in the tail vein. The next day,except for the blank group and the simple IgE group,the ears of the other groups were smeared with 2 4-dinitrofluorobenzene 50 μl to activate for the establishment of the animal model of urticaria. On the 8 th day,the degranulation of skin mast cells,the expression of protease activated receptor 2( PAR-2) mRNA and serum histamine concentration were detected in each group. Results With the observation of microscope,there was no obvious degranulation of mast cells in the blank group and the simple IgE group. The degranulation of mast cells in the model group was obvious. The number of degranulation of mast cells in each administration group was lower than that in the model group. The number of degranulation in the Guominjian Decoction group was relatively small. Compared with the blank group and the simple Ig E group,the number of degranulation,PAR-2 mRNA and histamine expression in the model group were significantly increased( P < 0. 05 or P < 0. 01). Compared with the model group,the number of degranulation,PAR-2 mRNA and histamine expression in the Modified Guominjian Decoction group were significantly decreased( P < 0. 01),and were decreased more than those in the Guominjian Decoction group and the loratadine group( P < 0. 05 or P < 0. 01).Conclusion The Modified Guominjian Decoction may down-regulate the expression of PAR-2 in the skin,reduce the release of serum histamine,inhibit the degranulation of skin mast cells,stabilize mast cells to achieve the function of treating and preventing urticaria.
引文
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[6]李润祥,冯承恩,梁碧华,等. Anti-DNP IgE联合DNFB诱导免疫性接触性荨麻疹小鼠模型的建立[J].皮肤性病诊疗学杂志,2015,22(1):25-28,32.
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[9]刘永平.李耀谦教授运用过敏煎治疗过敏性疾病的经验[J].名医,2018(1):96-97.
[10]中华医学会皮肤性病学分会免疫学组.中国荨麻疹诊疗指南(2014版)[J].中华皮肤科杂志,2014,47(7):514-516.
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[12]AKIYAMA T,LERNER EA,CARSTENS E. Proteaseactivated receptors and itch[J]. Handb Exp Pharmacol,2015,226:219-235. doi:10. 1007/978-3-662-44605-8_13.
[13]PALMER HS,KELSO EB,LOCKHART JC,et al. Proteaseactivated receptor 2 mediates the proinflammatory effects of synovial mast cells[J]. Arthritis Rheum,2007,56(11):3532-3540.
[14]ISHIKAWA C,TSUDA T,KONISHI H,et al. Tetracyclines modulate protease-activated receptor 2-mediated proinflammatory reactions in epidermal keratinocytes[J].Antimicrob Agents Chemother,2009,53(5):1760-1765.
[15]吕静,刘娟娟,廖晓容.类胰蛋白酶在慢性荨麻疹发病中的作用[J].临床皮肤科杂志,2013,42(3):197-199.
[16]李慧,刘玉峰,李巍.荨麻疹及其亚型的诊断和治疗进展[J].中国美容医学,2014,23(7):594-597.
[17]HE SH. Key role of mast cells and their major secretory products in inflammatory bowel disease[J]. World J Gastroenterol,2004,10(3):309-318.
[18]谢华,何韶衡,傅意玲.蛋白酶激活受体-2激动剂对人皮肤和扁桃体肥大细胞释放组胺和类胰蛋白酶的影响[J].医学争鸣,2005,26(7):599-602.
[19]TSUJII K,ANDOH T,LEE JB,et al. Activation of proteinase-activated receptors induces itch-associated response through histamine-dependent and independent pathways in mice[J]. J Pharmacol Sci,2008,108(3):385-388.
[2]宋红娟,王宝娟,李秋梅,等.灭荨汤联合枸地氯雷他定片治疗慢性荨麻疹疗效观察[J].中国中西医结合皮肤性病学杂志,2018,17(1):45-47.
[3]龙声志,吴贤波.“以皮治皮”理论源流及研究进展[J].中医杂志,2017,58(24):2147-2150.
[4]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,2011:1261-1264.
[5]李润祥,冯承恩,梁碧华,等.小鼠免疫性接触性荨麻疹动物模型的构建[J].中华皮肤科杂志,2015,48(6):421-425.
[6]李润祥,冯承恩,梁碧华,等. Anti-DNP IgE联合DNFB诱导免疫性接触性荨麻疹小鼠模型的建立[J].皮肤性病诊疗学杂志,2015,22(1):25-28,32.
[7]欧阳恒.论难治性皮肤病之诊治[J].湖南中医药大学学报,2007,27(4):4-6.
[8]景冰,贾颖.过敏煎加味治疗瘾疹临床经验举隅[J].山西中医学院学报,2017,18(6):63-64.
[9]刘永平.李耀谦教授运用过敏煎治疗过敏性疾病的经验[J].名医,2018(1):96-97.
[10]中华医学会皮肤性病学分会免疫学组.中国荨麻疹诊疗指南(2014版)[J].中华皮肤科杂志,2014,47(7):514-516.
[11]MOORMANN C,ARTUC M,POHL E,et al. Functional characterization and expression analysis of the proteinaseactivated receptor-2 in human cutaneous mast cells[J]. J Invest Dermatol,2006,126(4):746-755.
[12]AKIYAMA T,LERNER EA,CARSTENS E. Proteaseactivated receptors and itch[J]. Handb Exp Pharmacol,2015,226:219-235. doi:10. 1007/978-3-662-44605-8_13.
[13]PALMER HS,KELSO EB,LOCKHART JC,et al. Proteaseactivated receptor 2 mediates the proinflammatory effects of synovial mast cells[J]. Arthritis Rheum,2007,56(11):3532-3540.
[14]ISHIKAWA C,TSUDA T,KONISHI H,et al. Tetracyclines modulate protease-activated receptor 2-mediated proinflammatory reactions in epidermal keratinocytes[J].Antimicrob Agents Chemother,2009,53(5):1760-1765.
[15]吕静,刘娟娟,廖晓容.类胰蛋白酶在慢性荨麻疹发病中的作用[J].临床皮肤科杂志,2013,42(3):197-199.
[16]李慧,刘玉峰,李巍.荨麻疹及其亚型的诊断和治疗进展[J].中国美容医学,2014,23(7):594-597.
[17]HE SH. Key role of mast cells and their major secretory products in inflammatory bowel disease[J]. World J Gastroenterol,2004,10(3):309-318.
[18]谢华,何韶衡,傅意玲.蛋白酶激活受体-2激动剂对人皮肤和扁桃体肥大细胞释放组胺和类胰蛋白酶的影响[J].医学争鸣,2005,26(7):599-602.
[19]TSUJII K,ANDOH T,LEE JB,et al. Activation of proteinase-activated receptors induces itch-associated response through histamine-dependent and independent pathways in mice[J]. J Pharmacol Sci,2008,108(3):385-388.