摘要
目的:探究青霉素对银屑病模型小鼠肠道菌群变化及免疫功能调节作用的影响。方法:24只4~6周龄雌性BALB/c小鼠随机分为青霉素组(备皮后外涂5%咪喹莫特软膏诱导银屑病模型,腹腔注射青霉素)、模型组(备皮后外涂5%咪喹莫特软膏诱导银屑病模型,腹腔注射生理盐水)和对照组(备皮后外涂凡士林软膏,腹腔注射生理盐水),每组8只,皮损面积和严重程度指数(PASI)评分和HE染色评估各组小鼠皮损情况,流式细胞术检测小鼠血清免疫细胞因子CD4^(+)T细胞、CD8^(+)T细胞、CD11c^(+)树突状细胞(DCs)水平,ELISA检测血清炎症细胞因子IL-17A、IL-17F、IL-23、IL-22水平,并分析肠道菌群变化。结果:对照组小鼠背部皮肤无任何明显变化,模型组小鼠第2天起,背部皮肤出现红斑、鳞片和浸润,并逐渐明显,第7天,皮肤表现与人斑块型银屑病相似,病理可见皮肤明显角化、棘突及炎症细胞浸润,青霉素组小鼠背部皮肤仅可见少量红斑和薄鳞片,无明显浸润,皮肤病理变化明显减轻;模型组第2、4、7天PASI评分明显高于对照组(P<0.05),青霉素组第2、4、7天PASI评分明显低于模型组(P<0.05);模型组CD4^(+)T细胞、CD8^(+)T细胞百分比明显低于对照组(P<0.05),CD11c^(+)DCs百分比明显高于对照组(P<0.05),青霉素组CD4^(+)T细胞百分比明显高于模型组(P<0.05),CD11c^(+)DCs百分比明显低于模型组(P<0.05),CD8^(+)T细胞百分比与模型组差异无统计学意义(P>0.05);模型组IL-17A、IL-17F、IL-22、IL-23水平明显高于对照组(P<0.05),青霉素组IL-17A、IL-17F、IL-22、IL-23水平明显低于模型组(P<0.05);模型组乳酸杆菌、双歧杆菌、肠球菌及肠杆菌数与对照组差异无统计学意义(P>0.05),青霉素组乳酸杆菌、双歧杆菌数明显少于模型组(P<0.05),肠球菌、肠杆菌数与对照组差异无统计学意义(P>0.05),青霉素组OTU、Chao1、ACE、Shannon值明显低于模型组(P<0.05),Simpson值明显高于模型组(P<0.05);模型组放线菌门、厚壁菌门、变形菌门及拟杆菌门比例与对照组差异无统计学意义(P>0.05),青霉素组放线菌门、变形菌门比例明显高于模型组(P<0.05),厚壁菌门、拟杆菌门比例明显低于模型组(P<0.05);模型组与对照组副拟杆菌属、粪芽孢菌属差异无统计学意义(P>0.05);青霉素组副拟杆菌属、粪芽孢菌属比例明显高于模型组(P<0.05)。结论:青霉素可通过改变肠道菌群组成,降低肠道微生物多样性,调节免疫细胞因子及相关因子表达,从而改善咪喹莫特诱导的小鼠银屑病样皮肤炎症,为临床确定银屑病治疗靶点提供参考。
Objective:To investigate effects of penicillin on intestinal flora changes and immune function regulation in model mice with psoriasis.Methods:24 female BALB/c mice aged 4~6 weeks were randomly divided into penicillin group(after skin preparation,external application of 5%imiquimod ointment for inducing psoriasis model,and intraperitoneal injection of penicillin),model group(after skin preparation,external application of 5%imiquimod ointment for inducing psoriasis model,and intraperitoneal injection of saline)and control group(after skin preparation,external application of vaseline ointment,and intraperitoneal injection of saline),8 cases in each group.Percutaneous area and severity index(PASI)score and HE staining to evaluate skin lesions of mice in each group.Flow cytometry to detect levels of serum immune cytokines CD4^(+)T cells,CD8^(+)T cells,CD11c+dendritic cells(DCs),and ELISA to detect serum inflammatory cytokines IL-17A,IL-17F,IL-23,IL-22 levels,and changes of intestinal flora were analyzed.Results:There was no obvious change in back skin of mice in control group.From 2nd d,erythema,scales and infiltration appeared on back skin and gradually became obvious in model group,and at 7 d,skin appearance was similar to human plaque psoriasis,and pathology showed obvious skin keratinization,spinous processes and inflammatory cell infiltration.Back skin of mice in penicillin group showed only a small amount of erythema and thin scales without obvious infiltration,and pathology showed that skin changes were significantly reduced.PASI scores in model group at 2,4,and 7 d were significantly higher than those in control group(P<0.05),and PASI scores in penicillin group at 2,4,and 7 d were significantly lower than those in model group(P<0.05).Percentages of CD4^(+)T cells and CD8^(+)T cells in model group were significantly lower than those in control group(P<0.05),while percentage of CD11c+DCs was significantly higher than that in control group(P<0.05),and percentage of CD4^(+)T cells in penicillin group was significantly higher than that in model group(P<0.05),while percentage of CD11c+DCs was significantly lower than that in model group(P<0.05),and there was no significant difference in percentage of CD8^(+)T cells(P>0.05).Levels of IL-17A,IL-17F,IL-22,and IL-23 in model group were significantly higher than those in control group(P<0.05),and levels of IL-17A,IL-17F,IL-22 and IL-23 in penicillin group were significantly lower than those in model group(P<0.05).There were no significant differences in numbers of Lactobacilli,Bifidobacteria,Enterococci and Enterobacter in model group and control group(P>0.05),and numbers of Lactobacilli and Bifidobacteria in penicillin group were significantly less than those in model group(P<0.05),and there were no signiifcant differences in numbers of Enterococci and Enterobacter compared with model group(P>0.05).Values of OTU,Chao1,ACE and Shannon in penicillin group were significantly lower than those in model group(P<0.05),while Simpson value was significantly higher than that in model group(P<0.05).Proportions of Actinobacteria,Firmicutes,Proteobacteria and Bacteroidetes in model group were not significantly different from those in control group(P>0.05),proportions of Actinobacteria and Proteobacteria in penicillin group were significantly higher than those in model group(P<0.05),while proportions of Firmicutes and Bacteroidetes were significantly lower than those in model group(P<0.05).There were no significant differences between model group and control group in Parabacteroides and Coprobacillus(P>0.05).Proportions of Parabacteroides and Coprobacillus in penicillin group were significantly higher than those in model group(P<0.05).Conclusion:Penicillin can improve psoriasis-like skin inflammation in mice induced by imiquimod by changing composition of intestinal flora,reducing diversity of intestinal microbes and regulating expressions of immune cytokines and related factors,which can provide reference for clinical determination of psoriasis treatment targets.
作者
金曌
王飞
苏慧
张峻
殷翘
JIN Zhao;WANG Fei;SU Hui;ZHANG Jun;YIN Qiao(Dermatology Department,Wuhan No.1 Hospital,Wuhan 430022,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第15期1814-1819,共6页
Chinese Journal of Immunology
基金
湖北省卫生健康委科研项目(WJ2019M025)。
关键词
青霉素
银屑病
免疫功能
肠道菌群
炎症因子
Penicillin
Psoriasis
Immune function
Intestinal flora
Inflammatory factors
作者简介
金曌,女,硕士,住院医师,主要从事皮肤性病方面的研究;通信作者:王飞,E-mail:DAGANGMAOa@163.com。
