针灸对湿热蕴肤型湿疹豚鼠皮肤屏障功能的影响

Effect of acupuncture and moxibustion on skin barrier function in guinea pigs with eczema of skin damp-heat accumulation

目的:观察针灸对湿热蕴肤型湿疹模型豚鼠皮肤角质层含水量、血清炎性因子及皮肤、肺、直肠组织水通道蛋白3(AQP_(3))表达的影响,探究针灸调控皮肤屏障功能的可能机制。方法:将24只雄性白化豚鼠随机分为空白组(6只)和造模组(18只),造模组采用2,4-二硝基氯苯(DNCB)诱导湿热蕴肤型湿疹模型,造模成功后的豚鼠随机分为模型组、西药组、针灸组,每组6只。西药组予氯雷他定片(0.8 mg/kg)灌胃,每日1次,连续7 d;针灸组针刺“肺俞”“脾俞”“曲池”“足三里”“血海”,留针15 min,同时于“肺俞”“足三里”施雀啄灸,每穴10 min,每日1次,连续7 d。分别于干预前后评定各组豚鼠湿疹面积及严重度指数(EASI)评分,应用皮肤测试仪检测皮肤角质层含水量(WCSC)及经皮肤水分流失量(TEWL);干预结束后,采用HE染色法观察各组豚鼠皮肤组织形态学变化,ELISA法测定各组豚鼠血清免疫球蛋白E(IgE)、白介素(IL)-4、IL-17含量,Western blot法测定各组豚鼠皮肤、肺、直肠组织AQP_(3)蛋白表达。结果:干预前,与空白组比较,其他各组豚鼠EASI评分、TEWL升高(P<0.01),WCSC降低(P<0.01);干预后,与模型组比较,西药组和针灸组豚鼠EASI评分、TEWL降低(P<0.05,P<0.01),WCSC升高(P<0.01)。空白组豚鼠表皮结构完整,真皮层纤维排列整齐;模型组豚鼠表皮角化过度,颗粒层、棘细胞层及基底层细胞增生,真皮层纤维排列紊乱,炎性细胞浸润;西药组、针灸组豚鼠形态学表现较模型组改善。与空白组比较,模型组豚鼠血清IgE、IL-17含量升高(P<0.01),血清IL-4含量及皮肤、肺、直肠组织AQP_(3)蛋白表达降低(P<0.01,P<0.05);与模型组比较,西药组、针灸组豚鼠血清IgE、IL-17含量降低(P<0.01),血清IL-4含量及针灸组豚鼠皮肤、肺、直肠组织AQP_(3)蛋白表达升高(P<0.01,P<0.05);与西药组比较,针灸组豚鼠血清IgE、IL-17含量升高(P<0.01),血清IL-4含量降低(P<0.01)。结论:针灸可改善湿热蕴肤型湿疹模型豚鼠表皮水分代谢情况和皮肤组织形态,其机制可能与调控炎性因子,上调AQP_(3)表达,进而修复皮肤屏障功能有关。

Objective To observe the effect of acupuncture and moxibustion on the water content of stratum corneum(WCSC),expression of serum inflammatory factors and aquaporin 3(AQP_(3))in skin,lung and rectum in guinea pigs with eczema of skin damp-heat accumulation,and to explore the possible mechanism of acupuncture and moxibustion for regulating skin barrier function.Methods A total of 24 male albino guinea pigs were randomly divided into a blank group(n=6)and a modeling group(n=18).The guinea pigs in the modeling group were induced by 2,4-dinitrochlorobenzene(DNCB)to establish the eczema model of skin damp-heat accumulation.The guinea pigs with successful modeling were further randomly divided into a model group,a medication group and an acupuncture-moxibustion group,6 guinea pigs in each group.The guinea pigs in the medication group were treated with loratadine tablets(0.8 mg/kg)by gavage,once a day for 7 days;the guinea pigs in the acupuncture-moxibustion group were treated with acupuncture at"Feishu"(BL 13),"Pishu"(BL 20),"Quchi"(LI 11),"Zusanli"(ST 36)and"Xuehai"(SP 10);at the same time,moxibustion was applied at"Feishu"(BL 13)and"Zusanli"(ST 36),moxibustion intervention for 10 min and needle retaining for 15 min at each acupoint,once a day for 7 days.The eczema area and severity index(EASI)score was evaluated before and After intervention,and WCSC and trans-epidermal water loss(TEWL)were measured by skin tester.After intervention,The HE staining was used to observe the changes of skin histomorphology in each group;ELISA was used to measure the contents of serum immunoglobulin E(IgE),interleukin(IL)-4 and IL-17;Western blot was used to measure the protein expression of AQP_(3) in skin,lung and rectum.Results Before the intervention,compared with the blank group,the EASI scores and TEWL were increased in the remaining groups(P<0.01),and the WCSC was decreased(P<0.01).After the intervention,compared with the model group,the EASI scores and TEWL were decreased(P<0.05,P<0.01),and WCSC was increased(P<0.01)in the medication group and the acupuncture-moxibustion group.The epidermal structure in the blank group was complete and the fibers in the dermis were arranged orderly;in the model group,epidermal hyperkeratosis,proliferation of granular layer,spinous cell layer and basal layer,and disordered arrangement of dermal fibers and infiltration of inflammatory cells were observed.The morphological performance in the medication group and the acupuncture-moxibustion group was better than that in the model group.Compared with the blank group,the contents of serum IgE and IL-17 were increased(P<0.01),and the content of serum IL-4 and the protein expression of AQP_(3) in skin,lung and rectum were decreased in the model group(P<0.01,P<0.05).Compared with the model group,the contents of serum Ig E and IL-17 were decreased and the contents of serum IL-4 were increased in the medication group and the acupuncture-moxibustion group(P<0.01),and the protein expression of AQP_(3) in skin,lung and rectum in the acupuncturemoxibustion group were increased(P<0.05).Compared with the medication group,the contents of serum IgE and IL-17 were increased(P<0.01),and the content of serum IL-4 was decreased(P<0.01)in the acupuncture-moxibustion group.Conclusion Acupuncture and moxibustion could improve the epidermal water metabolism and skin tissue morphology in guinea pigs with eczema of skin damp-heat accumulation.Its mechanism may be related to regulating inflammatory factors,up-regulating the expression of AQP_(3),and then repairing the skin barrier function.