含不同剂量酸枣仁的敷和汤对特应性皮炎模型小鼠IgE水平及肠道菌群的影响

Study on the Effect of Fuhe Decoction(敷和汤)with Different Doses of Suanzaoren(Ziziphus jujuba)on IgE Levels and Intestinal Flora in Atopic Dermatitis Model Mice

目的探讨含不同剂量酸枣仁的敷和汤治疗特应性皮炎的作用机制。方法48只雌性BALB/c小鼠随机分为正常组,模型组,氯雷他定组,含高、中、低剂量酸枣仁的敷和汤组,每组8只。除正常组外,其余各组均用2'4-二硝基氟苯(DNFB)连续刺激制备特应性皮炎模型。造模后含高、中、低剂量酸枣仁的敷和汤组分别给予敷和汤24、18、15 g/(kg·d)灌胃,氯雷他定组给予氯雷他定干混悬剂0.001 g/(kg·d)灌胃,正常组和模型组给予10 ml/(kg·d)生理盐水灌胃,各组均灌胃14天。分别于造模第7、14天和给药第7、14天观察小鼠10 min内搔抓次数和皮损评分;ELISA法检测血清中免疫球蛋白E(IgE)含量;HE染色观察皮肤组织病理形态学改变;对小鼠结肠中粪便进行16S rRNA基因测序检测肠道菌群多样性及丰度变化。结果与正常组比较,造模第7、14天和灌胃第7、14天模型组小鼠10 min内搔抓次数增加且皮损评分升高(P<0.05),灌胃后小鼠表皮角化过度或不全、棘层细胞明显增厚、大量炎性细胞浸润;血清中IgE含量升高(P<0.05),厚壁菌门丰度降低,拟杆菌门、细菌丰度升高,乳杆菌属和普雷沃菌属相对丰度降低,拟杆菌属和颤螺菌属相对丰度升高(P<0.05)。与模型组比较,灌胃第7、14天氯雷他定组和含高、中剂量酸枣仁的敷和汤组小鼠10 min内搔抓次数和皮损评分均降低(P<0.05),血清中IgE含量降低,含中剂量酸枣仁的敷和汤组厚壁菌门丰度升高、拟杆菌门丰度降低,氯雷他定组拟杆菌属丰度降低,含中剂量酸枣仁的敷和汤组拟杆菌属丰度降低、乳杆菌属和普雷沃菌属丰度均升高(P<0.05)。与氯雷他定组比较,灌胃第7、14天含低剂量酸枣仁的敷和汤组小鼠皮损评分升高,含高、低剂量酸枣仁的敷和汤组小鼠搔抓次数增多;灌胃后含低剂量酸枣仁的敷和汤组小鼠IgE含量升高,含中剂量酸枣仁的敷和汤组厚壁菌门升高、拟杆菌门降低,乳杆菌属和普雷沃菌属升高,含高剂量酸枣仁的敷和汤组拟杆菌属升高(P<0.05)。结论含不同剂量酸枣仁的敷和汤能改善特应性皮炎的临床症状,调节肠道菌群的相对丰度纠正菌群的紊乱情况,其中以含中剂量酸枣仁的敷和汤组效果最优,且与氯雷他定组效果相当,降低血清中的IgE炎症反应可能是其作用机制之一。

Objective To investigate the effect of Fuhe Decoction(敷和汤)with different doses of Suanzaoren(Ziziphus jujuba)for atopic dermatitis(AD).Methods Forty-eight female BALB/c mice were randomly divided into normal group,model group,loratadine group,and Fuhe Decoction groups with high,medium,and low doses of Fuhe Decoction(Fuhe Decoction high-,medium-,and low-dose groups),with eight mice in each group.The AD model was pre⁃pared by continuous stimulation with 2,4-dinitrofluorobenzene(DNFB)in all groups but the normal group.After mod⁃elling,the Fuhe Decoction high-,medium-and low-dose groups were given 24,18 and 15 g(/kg·d)of Fuhe Decoc⁃tion,the loratadine group was given 0.001 g(/kg·d)of loratadine dry suspension,and the normal group and the model group were given 10 ml(/kg·d)of normal saline by gavage.All groups were gavaged for 14 days.The number of scratches within 10 min and the score of skin lesions were observed on the 7th and 14th days of modelling and on the 7th and 14th days of drug administration,respectively;serum immunoglobulin E(IgE)was detected by ELISA;the histopathological and morphological changes of the skin were observed by HE staining;and the diversity and abundance of intestinal flora were detected by 16S rRNA sequencing of fecal matter from the colon of the mice.Results Compared with the normal group,mice in the model group on the 7th day and the 14th day of modelling and the 7th day,the 14th day of gavage showed increased scratching within 10 min and higher skin lesion scores(P<0.05),with hyper⁃keratotic or incomplete epidermis,marked thickening of spiny cells,and a large number of inflammatory cells infiltrated in the mice after gavage;serum levels of IgE elevated(P<0.05),and the abundance of Bacillota decreased,that of the Bacteroidota and bacteria elevated,and relative abundance of Lactobacillus spp.and Prevotella spp.decreased,and relative abundance of Anaplasma spp.and Treponema spp.increased(P<0.05).Compared with the model group,the number of scratches within 10 min and the skin lesion scores of mice in the loratadine group and the Fuhe Decoction medium-and high-dose groups decreased on the 7th day and the 14th day of gavage(P<0.05),serum IgE reduced,and the bacteria reduced in the loratadine group,the abundance of Bacteroidesmus spp.increased and Bac⁃teriodesmus spp.decreased in the medium-dose group of Fuhe Decoction,the abundance of Bacteriodesmus spp.decreased in the loratadine group,the abundance of Bacteriodesmus spp.decreased,and that of both Lactobacillus spp.and Prevotella spp.increased in Fuhe Decoction medium-dose group(P<0.05).Compared with the loratadine group,the skin lesion scores increased in Fuhe Decoction low-dose group,and the number of scratching increased in the Fuhe Decoction low-and high-dose groups on the 7th day and the 14th day of gavage;the IgE content increased in Fuhe Decoction low-dose group,the Bacillota increased and the Bacteroidota decreased,the Lactobacillus spp.and Prevotella spp.increased in Fuhe Decoction middle-dose group,and Anopheles spp.increased in Fuhe Decoction highdose group after gavage(P<0.05).Conclusion Fuhe Decoction can improve the clinical symptoms of AD,regulate the relative abundance of intestinal flora to correct the disorders of the bacterial flora,among which the effect of Fuhe Decoction medium-dose group is optimal and comparable to that of the loratadine group,and the reduction of serum IgE inflammatory response may be one of its mechanisms of action.