艾灸对银屑病小鼠皮损表现和免疫炎性反应的影响

Effect of moxibustion on skin lesions and immune inflammatory response in psoriasis mice

目的:观察艾灸对银屑病小鼠皮损改善作用和对免疫炎性反应的影响,探讨艾灸治疗银屑病的可能机制。方法:将32只雄性BALB/c小鼠随机分为正常组、模型组、艾灸组和西药组,每组8只。模型组、艾灸组和西药组采用背部涂抹5%咪喹莫特乳膏7 d诱导银屑病小鼠模型。造模同时,艾灸组予背部皮损悬灸治疗,每日1次,每次20 min;西药组予1 mg/kg甲氨蝶呤片溶液灌胃给药,每日1次,两组均连续干预7 d。观察各组小鼠每日皮损变化情况,并评定其银屑病皮损面积和疾病严重程度指数(PASI)评分;HE染色法观察小鼠皮损组织形态学变化;免疫组化法检测皮损增殖细胞核抗原(PCNA)、T淋巴细胞表面标志CD3阳性表达情况;ELISA法检测血清白介素(IL)-17A表达水平,实时荧光定量PCR法检测皮损组织肿瘤坏死因子-α(TNF-α)、IL-1β和IL-6 mRNA相对表达。结果:模型组小鼠背部鳞屑增多、肥厚,皮肤干燥,表皮红肿,浸润明显,PASI各项评分与总分均高于正常组(P<0.01);艾灸组小鼠PASI鳞屑、浸润评分和总分低于模型组(P<0.01),西药组小鼠PASI浸润评分和总分低于模型组(P<0.01,P<0.05)。模型组小鼠炎性细胞浸润明显,与正常组比较,表皮层厚度增加(P<0.01);艾灸组、西药组小鼠炎性细胞浸润和Munro微脓肿减少,与模型组比较,表皮层厚度减少(P<0.01)。与正常组比较,模型组小鼠PCNA阳性、CD_(3)^(+)T细胞数增多(P<0.01),体质量减轻、脾指数增加(P<0.01),血清IL-17A含量及皮损组织TNF-α、IL-1β和IL-6 mRNA相对表达量升高(P<0.01)。与模型组比较,艾灸组、西药组小鼠PCNA阳性、CD_(3)^(+)T细胞数减少(P<0.01),脾指数及TNF-α、IL-1β和IL-6mRNA相对表达量降低(P<0.01);艾灸组小鼠体质量高于模型组(P<0.01);西药组小鼠血清IL-17A含量低于模型组(P<0.01)。艾灸组小鼠TNF-α和IL-1βmRNA相对表达量高于西药组(P<0.01)。结论:艾灸可有效改善银屑病小鼠皮损鳞屑和浸润表现,其机制可能与抑制炎性反应、调节免疫功能有关。

Objective To observe the effect of moxibustion on skin lesions and immune inflammatory response in psoriasis mice,and to explore the possible mechanism of moxibustion for psoriasis.Methods A total of 32 male BALB/c mice were randomly divided into a normal group,a model group,a moxibustion group and a medication group,8 mice in each group.Psoriasis model was induced by applying 5%imiquimod cream on the back for 7 days in the model group,the moxibustion group and the medication group.At the same time of model establishment,the moxibustion group was treated with suspension moxibustion on skin lesions on the back,20 min each time,once a day;the medication group was treated with 1 mg/kg methotrexate tablet solution by gavage,once a day.Both groups were intervened for 7 days.The daily changes of skin lesions were observed,and the psoriasis area and severity index(PASI)score was evaluated;the histopathological changes of skin lesions were observed by HE staining;the positive expression of proliferating cell nuclear antigen(PCNA)and T lymphocyte surface marker CD3 were detected by immunohistochemistry;the expression level of serum interleukin(IL)-17 A was detected by ELISA,and the relative expressions of tumor necrosis factor-α(TNF-α),IL-1βand IL-6 mRNA in skin lesions were detected by real-time PCR.Results The increased and hypertrophy scale,dry skin,red and swollen epidermis and obvious infiltration were observed in the model group,and each score and total score of PASI were higher than those in the normal group(P<0.01).The scale score,infiltration score,and total score of PASI in the moxibustion group were lower than those in the model group(P<0.01);the infiltration score and total score of PASI in the medication group were lower than those in the model group(P<0.01,P<0.05).The inflammatory cell infiltration in the model group was obvious,and the thickness of epidermal layer was increased compared with that in the normal group(P<0.01);the inflammatory cell infiltration and Munro micro abscess were decreased in the moxibustion group and the medication group,and the thickness of epidermal layer was decreased compared with that in the model group(P<0.01).Compared with the normal group,the positive cell number of PCNA and CD_(3)^(+)T was increased(P<0.01),and the body mass was decreased,and the spleen index was increased(P<0.01),and the expression of serum IL-17 A and the relative expression of TNF-α,IL-1βand IL-6 m RNA in the skin lesions was increased in the model group(P<0.01).Compared with the model group,the positive cell number of PCNA and CD_(3)^(+)T was reduced(P<0.01),and the spleen index and the relative expression of TNF-α,IL-1βand IL-6 m RNA were reduced(P<0.01)in the moxibustion group and the medication group;the body mass of mice in the moxibustion group was higher than that in the model group(P<0.01);the content of serum IL-17 A in the medication group was lower than that in the model group(P<0.01);the relative expression of TNF-α,IL-1βm RNA in the moxibustion group was higher than that in the medication group(P<0.01).Conclusion Moxibustion could effectively improve the scale and infiltration of skin lesions in psoriasis mice.Its mechanism may be related to inhibiting inflammatory response and regulating immunity.