肤痒清凝胶通过调控p38 MAPK/JNK/Caspase-3信号通路改善DNCB诱导的小鼠特应性皮炎*

作者：乔善鑫1，李方睿2，禹亚杰3，魏海燕4，徐 萌5，王 英1，徐兴华1，马亚琴1

单位：1.内蒙古包钢医院，内蒙古 包头 014010； 2.国药北方医院，内蒙古 包头 014010； 3.四川护理职业学院，四川 成都 610100； 4.上海市浦东新区南汇精神卫生中心，上海 200120； 5.四川省中西医结合医院，四川 成都 610100

引用：引用：乔善鑫，李方睿，禹亚杰，魏海燕，徐萌，王英，徐兴华，马亚琴.肤痒清凝胶通过调控p38 MAPK/JNK/Caspase-3信号通路改善DNCB诱导的小鼠特应性皮炎[J].中医药导报,2025,31(6):55-60.

DOI：10.13862/j.cn43-1446/r.2025.06.010

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摘要：目的：探究肤痒清凝胶（FYQO）调控p38丝裂原活化蛋白激酶（p38 MAPK）/c-Jun氨基末端激酶（JNK）/胱天蛋白酶-3（Caspase-3）信号通路对2,4-二硝基氯苯（DNCB）诱导的小鼠特应性皮炎（AD）皮损的改善作用。方法：75只小鼠随机分为对照组（Control组）、模型组（DNCB组）、肤痒清凝胶低剂量组（FYQO-1 g/kg组）、肤痒清凝胶中剂量组（FYQO-2 g/kg组）、肤痒清凝胶高剂量组（FYQO-4 g/kg组），每组15只。实验首日，小鼠腹部涂抹30 μL 5.0% DNCB溶液致敏，第3天于背部涂50 μL 0.2%DNCB溶液二次激发，此后每3 d重复1次，共计3次，第9天观察背部皮肤反应，若出现红肿、丘疹等即为造模成功。Control组不处理，DNCB组涂抹空白凝胶，FYQO-1 g/kg组、FYQO-2 g/kg组、FYQO-4 g/kg组分别涂抹1、2、4 g/kg肤痒清凝胶，2次/d，共9 d；在首次涂抹凝胶后的第3天和第9天观察小鼠搔抓行为，采用湿疹面积及严重程度指数（EASI）评分标准评价；苏木精-伊红（HE）染色观察皮损组织病理变化；免疫组织化学检测标志物蛋白表达；实时荧光定量聚合酶链式反应（RT-qPCR）和免疫印迹检测p38 MAPK/JNK/Caspase-3通路相关标志物mRNA和蛋白水平。结果：在首次涂抹凝胶后第3、9天，与Control组比较，DNCB组小鼠搔抓次数显著增加，在停止涂抹凝胶当天及结束后1周，DNCB组EASI评分升高，小鼠背部皮肤出现AD样皮损，表皮和棘皮层增厚，炎性细胞浸润，P38、JNK和cleaved Caspase-3 mRNA和蛋白水平明显升高（P＜0.05）；在首次涂抹凝胶后第3、9天，与DNCB组比较，FYQO-1 g/kg、FYQO-2 g/kg、FYQO-4 g/kg组小鼠的搔抓次数显著减少，在停止涂抹凝胶当天及结束后1周，小鼠的EASI评分显著降低，小鼠背部皮肤出现不同程度的皮肤变薄、红斑和水肿，表皮层和棘皮层轻度增厚，炎症细胞浸润减少等情况，P38、JNK和cleaved Caspase-3 mRNA和蛋白水平显著降低（P＜0.05）；FYQO-4 g/kg组的效果优于FYQO-1 g/kg组和FYQO-2 g/kg组（P＜0.05）。结论：肤痒清凝胶可通过调控p38MAPK/JNK/Caspase-3信号通路改善DNCB诱导的小鼠特应性皮炎。

关键词：特应性皮炎；肤痒清凝胶；p38 MAPK/JNK/Caspase-3信号通路；2,4-二硝基氯苯；小鼠

Abstract：

Objective: To explore the effect of Fuyangqing Ointment (FYQO) on the improvement of skin lesions in mice with atopic dermatitis (AD) induced by 2,4-dinitrochlorobenzene (DNCB) by regulating p38 mitogen activated protein kinase (p38 MAP) /c-Jun terminal kinase (JNK)/cysteine protease-3 (Caspase-3) pathway. Methods: Totally 75 mice were randomly divided into control group, model group (DNCB group), low dose group of Fuyangqing Ointment (FYQO-1 g/kg group), medium dose group of Fuyangqing Ointment (FYQO-2 g/kg group), and high dose group of Fuyangqing Ointment (FYQO-4 g/kg group), with 15 mice in each group. On the 1st day of the experiment, 30 μL of 5.0% DNCB solution was applied to the abdomen of mice to induce sensitization. On the 3rd day, 50 μL of 0.2% DNCB solution was applied to the back for a second stimulation. This was repeated every 3 days for a total of 3 times. On the 9th day, the skin reaction on the back was observed. If redness, swelling, papules, etc. appeared, the model was considered successful. The control group was not treated, while the DNCB group was smeared with blank gel. The FYQO-1 g/kg group, FYQO-2 g/kg group and FYQO-4 g/kg group were smeared with 1, 2, and 4 g/kg Fuyangqing Ointment respectively, twice a day for 9 days. Scratching behavior of mice was observed on the 3rd and 9th days after the 1st application of gel, and the scratching behavior was evaluated with the eczema area and severity index (EASI) scoring standard. Hematoxylin eosin staining was used to observe the pathological changes of skin lesions, and immunohistochemical was used to detect marker protein expression. Real time quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to detect the mRNA and protein levels of p38MAPK/JNK/Caspase-3 pathway biomarkers. Results: On the 3rd and 9th day after the first application of gel, compared with the control group, the scratching times of mice increased in the DNCB group. On the day when the gel was stopped and one week after the end, the EASI score increased in the DNCB group, and the back skin of mice showed AD like lesions, thickening of epidermis and spinous cortex, and inflammatory cell infiltration. Compared with the control group, the mRNA and protein expression levels of P38, JNK and cleaved Caspase-3 were increased in DNCB group (P<0.05). On the 3rd and 9th days after the first application of gel, compared with the DNCB group, the scratching times of the mice reduced in FYQO-1 g/kg, FYQO-2 g/kg, and FYQO-4 g/kg groups. On the day when the gel was stopped and one week after the end, the EASI scores of the mice reduced in FYQO-1 g/kg, FYQO-2 g/kg, and FYQO-4 g/kg groups. The skin of the mice's back was thinner, erythema and edema to varying degrees, and the epidermis and spinous cortex were slightly thickened in FYQO-1 g/kg, FYQO-2 g/kg, and FYQO-4 g/kg groups. The inflammatory cell infiltration was reduced in FYQO-1 g/kg, FYQO-2 g/kg, and FYQO-4 g/kg groups. Compared with the DNCB group, the mRNA and protein expression of levels of P38, JNK, and cleaved Caspase-3 reduced in FYQO-1 g/kg, FYQO-2 g/kg, and FYQO-4 g/kg groups (P<0.05). The FYQO-4 g/kg group showed better effect than FYQO-2 g/kg and FYQO-4 g/kg groups (P<0.05). Conclusion: Fuyangqing Ointment can improve DNCB induced atopic dermatitis in mice by regulating p38 MAPK/JNK/Caspase-3 pathway.

Key words：atopic dermatitis; Fuyangqing Ointment; p38 MAPK/JNK/Caspase-3 pathway; 2,4-dinitrochlorobenzene; mouse

发布时间：2026-01-03

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