电针对荨麻疹大鼠皮下疏松结缔组织中肥大细胞Lyn、Syk表达的影响

Effect of electroacupuncture on expressions of Lyn and Syk in mast cells of subcutaneous loose connective tissue in rats with urticaria

目的:观察电针预处理对荨麻疹大鼠皮下疏松结缔组织中肥大细胞脱颗粒及其过程中酪氨酸激酶Lyn、脾酪氨酸激酶(Syk)表达的影响,探讨电针干预荨麻疹的潜在生物学机制。方法:32只SD大鼠随机分为空白组、模型组、电针组、阳性药物组,每组8只。除空白组外,其余3组大鼠均采用皮肤被动过敏反应(PCA)制备荨麻疹大鼠模型。电针组电针双侧"曲池""血海""足三里",疏密波,频率2 Hz/15 Hz,电流强度1 mA,每日1次,每次20 min,连续7 d;阳性药物组给予氯雷他定(1 mg×kg-1×d-1)灌胃,每日1次,连续7 d。各组大鼠完成PCA抗原攻击30min后取材进行指标检测。比色法检测大鼠致敏部位皮肤伊文斯蓝染料渗出量;HE染色法观察致敏部位皮肤组织形态改变;甲苯胺蓝染色法观察致敏部位皮下疏松结缔组织中肥大细胞脱颗粒情况;免疫组织化学法检测肥大细胞脱颗粒过程中Lyn、Syk蛋白的表达。结果:模型组大鼠致敏部位表皮层增厚,细胞排列层次紊乱,真皮层内可见大量炎性细胞浸润;阳性药物组、电针组致敏部位表皮层薄,细胞排列层次较清晰,炎性细胞浸润情况较模型组明显减轻。与空白组比较,模型组皮肤染料渗出量吸光度(OD)值、肥大细胞脱颗粒率、Lyn与Syk阳性表达均升高(P<0.01);与模型组比较,电针组和阳性药物组皮肤染料渗出量吸光度(OD)值、肥大细胞脱颗粒率、Lyn与Syk阳性表达均降低(P<0.01);与阳性药物组比较,电针组肥大细胞脱颗粒率显著升高(P<0.01)。结论:电针"曲池""血海""足三里"可通过调控荨麻疹大鼠肥大细胞脱颗粒,降低血管渗透性,发挥抗过敏作用,其机制可能与抑制肥大细胞Lyn、Syk蛋白表达有关。

Objective To observe the effect of electroacupuncture(EA)preconditioning on the expressions of tyrosine kinase Lyn and spleen tyrosine kinase(Syk)in mast cells of subcutaneous loose connective tissue in the rats with urticaria and explore the potential biological mechanism of EA in the intervention of urticaria.Methods A total of 32 SD rats were randomized into a blank group,a model group,an EA group and a positive medication group,8 rats in each one.Except of the blank group,the passive cutaneous anaphylaxis(PCA)was adopted to prepare the model of urticaria in the rats of the rest three groups.In the EA group,EA was applied to bilateral"Quchi"(LI 11),"Xuehai"(SP 10)and"Zusanli"(ST 36),with disperse-dense wave,2 Hz/15 Hz in frequency and 1 mA in current intensity,once daily,for 20 min each time,consecutively for 7 days.In the positive medication group,loratadine(1 mg·kg-1·d-1)was for intragastric administration,once daily,consecutively for 7 days.The samples were collected for index detection 30 min after PCA antigen challenge in the rats of each group.Spectrophotometer was adopted to determine the effusion quantity of Evans blue in the allergized site of skin.HE staining was used to observe the morphological changes in the allergized site of skin.Toluidine blue staining was provided to observe mast cell degranulation in subcutaneous loose connective tissue in the allergized site of skin.Immunohistochemistry was applied to determine the protein expressions of Lyn and Syk during degranulation of mast cells.Results In the rats of the model group,the eipdermis of allergized site was thickening,cells were disorganized in hierarchy and inflammatory cells were infiltrated largely in the dermis.In the positive medication group and the EA group,the epidermis was getting thin,cell arrangement was clear and the inflammatory cell infiltration was obviously alleviated as compared with the model group.Compared with the blank group,the OD value of skin dye effusion quantity,the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all increased in the model group(P<0.01).Compared with the model group,the OD value of skin dye effusion quantity,the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all reduced in the EA group and the positive medication group(P<0.01).Compared with the positive medication group,the degranulation rate of mast cells was increased significantly in the EA group(P<0.01).Conclusion Electroacupuncture at"Quchi"(LI 11),"Xuehai"(SP 10)and"Zusanli"(ST 36)reduces vascular permeability and gives play to the role of anti-allergy by the way of regulating and controlling the degranulation of mast cells in the rats with urticaria and the effect mechanism of electroacupuncture may be related to the inhibition of protein expressions of Lyn and Syk in mast cells.