文献类型：期刊文献

中文题名：基于“肺与大肠相表里”探究复方嗜酸乳杆菌片联合马齿苋提取液对湿疹大鼠肠道菌群、免疫功能及cAMP/PKA的影响

英文题名：Exploring effects of compound Lactobacillus acidophilus tablets combined with Portulacae Herba extract on intestinal microbiota abundance,immune function,and cAMP/PKA in eczema rats based on theory of"exterior?interior relationship between lung and large intestine"

作者：徐兰萍[1];梁恬[2];陈继军[3];张琪[2];谢泽初[1]

第一作者：徐兰萍

机构：[1]甘肃中医药大学附属医院皮肤科,甘肃兰州730000;[2]甘肃中医药大学附属医院小儿骨科,甘肃兰州730000;[3]兰州市疾病预防控制中心性病艾滋病防治科,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2025

卷号：59

期号：10

起止页码：66

中文期刊名：上海中医药杂志

外文期刊名：Shanghai Journal of Traditional Chinese Medicine

基金：甘肃省自然科学基金项目(24jRRA552)。

语种：中文

中文关键词：湿疹;复方嗜酸乳杆菌片;马齿苋提取液;肠道菌群;免疫功能;中西医结合疗法;大鼠模型

外文关键词：eczema;compound Lactobacillus acidophilus tablets;Portulacae Herba extract;gut microbiota;immune function;integrated traditional Chinese and Western medicine therapy;rat model

摘要：目的探究复方嗜酸乳杆菌片联合马齿苋提取液对湿疹大鼠肠道菌群、免疫功能及环腺苷酸(cAMP)/蛋白激酶A(PKA)的影响。方法成年雄性SD大鼠50只随机分为5组(n=10):正常组、模型组、复方嗜酸乳杆菌组、马齿苋组及联合组。除正常组外,其余各组均建立湿疹动物模型。造模成功后,复方嗜酸乳杆菌组灌胃复方嗜酸乳杆菌片,马齿苋组灌胃马齿苋提取液,联合组大鼠灌胃复方嗜酸乳杆菌片+马齿苋提取液,正常组和模型组大鼠灌胃等体积的0.9%氯化钠溶液,连续干预14 d。干预结束后:①观察大鼠背部皮损情况,记录搔抓次数;②采用酶联免疫吸附分析(ELISA)法检测外周血γ干扰素(IFN?γ)及白细胞介素(IL)-4水平;③16S rRNA法检测肠道菌群丰度;④采用苏木精-伊红(HE)染色法观察皮损组织病理形态;⑤Western blot法检测cAMP、PKA蛋白水平。结果①正常组大鼠皮肤为淡红色,细致滑嫩,质地柔软,皮肤纹理清晰可见。模型组大鼠背部皮损明显,可见皮肤红斑、浸润、鳞屑等,符合建模特征。与模型组比较,其余用药组大鼠背部皮损明显好转,以联合组改善最为显著。与正常组比较,模型组大鼠搔抓次数增多(P<0.05)。与模型组比较,复方嗜酸乳杆菌组、马齿苋组及联合组大鼠搔抓次数减少(P<0.05)。与复方嗜酸乳杆菌组及马齿苋组比较,联合组大鼠搔抓次数均减少(P<0.05)。②与正常组比较,模型组大鼠IL-4水平升高(P<0.05),IFN-γ水平降低(P<0.05)。与模型组比较,复方嗜酸乳杆菌组及马齿苋组大鼠IL-4水平降低(P<0.05),IFN-γ水平升高(P<0.05)。与复方嗜酸乳杆菌组及马齿苋组比较,联合组大鼠IL-4水平降低(P<0.05),IFN-γ水平升高(P<0.05)。③与正常组比较,模型组厚壁菌门丰度增加;与模型组比较,复方嗜酸乳杆菌组、马齿苋组及联合组厚壁菌门丰度均降低,其中以联合组厚壁菌门丰度最低。④HE染色可见:正常组大鼠表皮角化正常,真皮乳头层未见水肿和炎症细胞浸润;模型组大鼠存在表皮角化,且存在大量炎症细胞浸润,部分伴随纤维增生及血管增生;复方嗜酸乳杆菌组、马齿苋组及联合组大鼠表皮炎症细胞逐渐消失,皮肤结构逐渐清楚,联合组大鼠仅在真皮层可见到少许炎症,水肿消失,病理形态改善程度最佳。⑤与正常组比较,模型组大鼠cAMP、PKA蛋白水平降低(P<0.05)。与模型组比较,复方嗜酸乳杆菌组大鼠cAMP、PKA蛋白水平升高(P<0.05)。与复方嗜酸乳杆菌组比较,马齿苋组大鼠cAMP、PKA蛋白水平升高(P<0.05)。与复方嗜酸乳杆菌组及马齿苋组比较,联合组大鼠cAMP、PKA蛋白水平升高(P<0.05)。结论复方嗜酸乳杆菌片联合马齿苋提取液可改善湿疹大鼠肠道菌群紊乱,恢复T辅助细胞Th1/Th2免疫平衡,并通过激活cAMP/PKA信号通路而缓解皮损。
Objective To investigate the effects of compound Lactobacillus acidophilus tablets(CLAT)combined with Portulacae Herba extract(POE)on intestinal microbiota,immune function,and the cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)pathway in rats with eczema.Methods Fifty adult male SD rats were randomly divided into five groups(n=10 per group):normal control,model,CLAT,POE,and combination(CLAT+POE)groups.Eczema models were established in all groups except the normal control group.After successful modeling,interventions were administered via gavage for 14 days:rats in the CLAT group received CLAT suspension,those in the POE group received POE solution,those in the combination group received CLAT+POE,and rats in the normal control and model groups were given 0.9%NaCl solution.After intervention,the following assessments were conducted:①skin lesions on the back and scratching frequency of rats;②serum interferon-γ(IFN-γ)and interleukin-4(IL-4)levels via enzyme-linked immunosorbent assay(ELISA);③intestinal microbiota abundance via 16S rRNA sequencing;④histopathology of lesional skin via hematoxylin and eosin(HE)staining;⑤cAMP and PKA protein levels via Western blot.Results①The skin of normal group rats appeared light red,delicate,and smooth,with soft and clear skin texture.The back skin of model group rats showed obvious erythema,infiltration,scales,etc.,consistent with eczema modeling characteristics.Compared with the model group,skin lesions on the back of rats in the other treatment groups improved significantly,with the most prominent improvement in the combination group.The scratching frequency was decreased in the CLAT,POE,and combination groups compared with the model group(P<0.05),and was further reduced in the combination group compared with the CLAT or POE group(P<0.05).②Compared with the normal group,the model group exhibited increased IL-4 levels and decreased IFN-γlevels(P<0.05).Compared with the model group,IL-4 levels were decreased and IFN-γlevels were increased in the CLAT and POE groups(P<0.05);moreover,the combination group showed lower IL-4 levels and higher IFN-γlevels than the CLAT or POE group(P<0.05).③Model rats had higher Firmicutes abundance than normal rats;all treatment groups reduced Firmicutes,with the combination group showing the greatest efficacy.④HE staining revealed that rats in the normal group had normal epidermal keratinization,and no edema or inflammatory cell infiltration was observed in the dermal papilla layer.In contrast,model group rats exhibited abnormal epidermal keratinization and abundant inflammatory cell infiltration,partially accompanied by fibrosis and vascular proliferation.In the CLAT,POE,and combination groups,inflammatory cells in the epidermis gradually disappeared,and skin structure gradually normalized;notably,the combination group showed only slight dermal inflammation and no edema,presenting the best pathological improvement.⑤Compared with the normal group,cAMP and PKA protein levels were decreased in the model group(P<0.05).Compared with the model group,cAMP and PKA levels were increased in the CLAT group(P<0.05),further increased in the POE group compared with the CLAT group(P<0.05),and the combination group showed higher levels than the CLAT or POE group(P<0.05).Conclusion The combination of CLAT and POE alleviates eczema by modulating intestinal microbiota,restoring the T helper cell Th1/Th2 balance,and activating the cAMP/PKA pathway,thereby synergistically enhancing therapeutic effects.