沙棘果油对超抗原诱导的特应性皮炎小鼠激素抵抗的干预作用及机制
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篇名: 沙棘果油对超抗原诱导的特应性皮炎小鼠激素抵抗的干预作用及机制
TITLE: Intervention effect and mechanism of Hippophae rhamnoides oil on glucocorticoid resistance in superantigen-induced atopic dermatitis mice
摘要: 目的 研究沙棘果油对超抗原诱导的特应性皮炎(AD)小鼠激素抵抗的干预作用,并探讨其作用机制。方法将50只小鼠按照随机数字表法分为5组,即正常对照组(A组)、模型组(B组)、地塞米松干预组(阳性对照,C组)、沙棘果油干预组(D组)、地塞米松+沙棘果油干预组(E组),每组10只。除A组外,其余各组均采用2,4-二硝基氯苯+葡萄球菌肠毒素B法制备AD小鼠模型。自实验第7天起,分别使用地塞米松(1.5mg/kg)和/或沙棘果油(10mL/kg)对C、D、E组小鼠进行灌胃,每天1次,连续28d。末次给药后,采用苏木素-伊红染色法观察小鼠耳部皮肤组织的病理形态学变化,采用酶联免疫吸附测定法检测小鼠血清中免疫球蛋白E(IgE)、白细胞介素4(IL-4)水平,采用免疫荧光单标法检测小鼠耳部皮肤组织中糖皮质激素受体α(GRα)、GRβ阳性细胞数,采用Westernblot法检测小鼠耳部皮肤组织中GRα、GRβ蛋白及蛋白激酶B(AKT)/核糖体蛋白S6激酶1(S6K1)信号通路相关蛋白的表达情况。结果与B组比较,C、D、E组小鼠左耳部皮肤炎症表现明显减轻;D、E组小鼠血清中IgE、IL-4水平显著降低(P<0.05),GRα阳性细胞数显著增加(P<0.05),核蛋白中GRα蛋白表达水平升高(P<0.05),总蛋白中Gαi1、Gαi3、p-S6K1、p-AKT蛋白表达水平均显著降低(P<0.05);E组小鼠GRβ阳性细胞数显著减少(P<0.05),总蛋白中GRβ表达水平显著降低(P<0.05)。与C组比较,E组小鼠左耳部皮肤炎症表现趋于消退,血清中IgE、IL-4水平均显著降低(P<0.05);D、E组小鼠GRα阳性细胞数显著增加(P<0.05),GRβ阳性细胞数显著减少(P<0.05),核蛋白中GRα蛋白表达水平升高(P<0.05),总蛋白中GRβ、Gαi1、p-S6K1、p-AKT的蛋白表达水平及E组小鼠Gαi3的蛋白表达水平均显著降低(P<0.05)。结论沙棘果油对超抗原诱导的AD小鼠激素抵抗具有干预作用,该作用可能是通过抑制Gαi1/3诱导的AKT/S6K1信号通路而发挥的。
ABSTRACT: OBJECTIVE To study the intervention effect of Hippophae rhamnoides oil on glucocorticoid resistance in superantigen-induced atopic dermatitis (AD) mice,and to explore the mechanism of action. METHODS Fifty mice were randomly divided into 5 groups,i.e. normal control group (group A),model group (group B),dexamethasone intervention group (positive control,group C),H. rhamnoides oil intervention group (group D),dexamethasone+H. rhamnoides oil intervention group (group E),with 10 mice in each group. Except for group A,other groups were given 2,4-dinitrochlorobenzene+staphylococcal enterotoxin B to induce the AD mice model. Starting from the 7th day of the experiment,groups C,D and E were given dexamethasone (1.5 mg/kg) and/or H. rhamnoides oil (10 mL/kg) intragastrically,once a day,for 28 consecutive days. After the last medication,the pathomorphological changes of ear tissue were observed by 节作用。E-mail:57667478@qq.com HE staining; the serum levels of immunoglobulin E (IgE) and interleukin 4 (IL-4) were detected by enzyme-linked immunosorbent assay. Positive cell count of glucocorticoid receptor α (GRα) and GRβ in the ear tissue of mice was detected by tyramide signal amplification. The expressions of GRα protein,GRβ protein,and protein kinase B (AKT)/ribosomal protein S6 kinase 1,S6K1 (S6K1) signaling pathway-related proteins were determined by Western blot assay. RESULTS Compared with group B,the skin inflammation in the left ear of the mice was significantly reduced in groups C,D and E,the serum levels of IgE and IL-4 were decreased significantly in groups D and E (P< 0.05),while the number of GRα positive cells and GRα protein expression were increased significantly (P<0.05); the protein levels of G protein inhibitory subunit 1 (Gαi1),Gαi3,phosphorylated S6K1 (p-S6K1) and phosphorylated AKT (p-AKT) were decreased significantly (P<0.05); the number of GRβ positive cells and protein expression of GRβ was decreased significantly in group E(P<0.05). Compared with group C,the skin inflammation in the left ear of the mice was almost clear away in group E,the serum levels of IgE and IL-4 were decreased significantly (P<0.05); the number of GRα positive cells and GRα protein expression were increased significantly in groups D and E (P<0.05); the protein levels of GRβ,Gαi1,p-S6K1 and p-AKT were all decreased significantly in groups D and E(P<0.05); and protein level of Gαi3 was decreased significantly in group E (P<0.05). CONCLUSIONS H. rhamnoides oil has an intervention effect on superantigen-induced glucocorticoid resistance of AD mice,which may be exerted by inhibition of the Gαi1/3-induced AKT/S6K1 signaling pathway.
期刊: 2023年第34卷第17期
作者: 王欣欣;陈小艳;李思佳;张文莲;王昆;窦德强
AUTHORS: WANG Xinxin,CHEN Xiaoyan,LI Sijia,ZHANG Wenlian,WANG Kun,DOU Deqiang
关键字: 沙棘果油;超抗原;特应性皮炎;激素抵抗;蛋白激酶B/核糖体蛋白S6激酶1信号通路
KEYWORDS: Hippophae rhamnoides oil; superantigen; atopic dermatitis; glucocorticoid resistance; AKT/S6K1 signaling
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