龙牡汤对急性湿疹大鼠Th1/Th2平衡、肥大细胞及皮肤屏障蛋白表达的影响研究

Effects of Longmu Decoction on Th1/Th2 Balance,Mastcells and Expression of Skin Barrier Related Proteins in Acute Eczema Rats

目的观察龙牡汤对急性湿疹大鼠Th2活化的抑制作用及皮肤屏障相关蛋白表达的影响。方法SD大鼠32只随机分为4组,正常组、模型组、龙牡汤组、白芍总苷组,每组8只。除正常组外,其余组通过二硝基氯苯在大鼠背部皮肤建立急性湿疹模型。造模成功后,龙牡汤组灌服中药30 g/(kg·d),白芍总苷组给予白芍总苷胶囊70 mg/(kg·d),正常组、模型组均灌服等量蒸馏水,干预周期14 d。ELISA检测血浆中白细胞介素-2(IL-2)、白细胞介素-4(IL-4)及IgE水平,HE染色光镜下观察皮肤组织形态,甲苯胺蓝染色观察各组皮肤组织中浸润肥大细胞数量,免疫组化法检测各组皮肤组织中E-caderin、ZO-1以及Claudin3的表达。结果与正常组比较,模型组大鼠静脉血IL-2水平减少,IL-4及IgE水平增高(P<0.05);皮肤组织表现为表皮增生,棘层肥厚,角化不全及糜烂,真皮层血管扩张、水肿,炎细胞浸润,肥大细胞数量增多;E-caderin、ZO-1以及Claudin3的表达减少(P<0.05)。与模型组比较,龙牡汤组及白芍总苷组IL-4及IgE水平下调(P<0.05),表皮增生、糜烂及炎症浸润均有不同程度的改善,浸润肥大细胞减少,E-caderin和ZO-1表达增加(P<0.05)。结论龙牡汤组可减轻急性湿疹大鼠皮肤损伤,可能与调控Th1/Th2平衡,抑制Th2活化,减少肥大细胞产生,增加皮肤屏障相关蛋白的表达有关。

Objective:To observe the inhibitory effect of Longmu Decoction on Th2 activation and skin barrier related protein expression in rats with acute eczema,and to explore the possible mechanism.Methods:32 SD rats were randomly divided into 4 groups,i.e.,the normal group,model group,Longmu Decoction group,positive drug total glucosides group,8 rats in each group.Except for the normal group,the other groups established an acute eczema model on dorsal skin of rats by dinitrochlorobenzene.After successful modeling,the Longmu Decoction group was given 30 g/(kg·d)of traditional Chinese medicine,the positive drug group was given 70 mg/(kg·d)total glucosides capsules,and the normal group and model group were given the same amount of distilled water.The intervention period was 14 d.The levels of IL-2,IL-4 and IgE in plasma were detected by ELISA.The morphology of skin tissue was observed under light microscope with HE staining.The number of infiltrating mast cells in the skin tissue of each group was observed by toluidine blue staining.Expressions of E-caderin,ZO-1 and Claudin3were tested by immunohistochemical method.Results:Compared with normal group,the levels of IL-2 in venous blood of model group were decreased,while the levels of IL-4 and IgE were increased(P<0.05).The skin lesions were characterized by epidermal hyperplasia,spinous layer hypertrophy,keratosis insufficiency and erosion,dermal vascular dilation,edema,inflammatory cell infiltration.The number of mast cells increased.The expression of e-caderin,ZO-1 and Claudin3 decreased(P<0.05).Compared with model group,the levels of IL-4 and IgE in Longmu Decoction and positive drug group were down-regulated(P<0.05),epidermal hyperplasia and inflammatory infiltration were significantly improved.The number of mast cells decreased and the expressions of E-caderin and ZO-1 increased(P<0.05).Conclusion:Longmu Decoction can alleviate skin injury in acute eczema rats,which may be related to regulating Th1/Th2 balance,inhibiting Th2 activation and the number of mast cells,and increas-ing the expression of skin barrier related proteins.