维胺酯通过激活RBP4/RARγ途径缓解大鼠耳部痤疮表皮过度角化  

Viaminate Alleviates Hyperkeratosis of Ear Acne Epidermis in Rats by Activating RBP4/RARγ Pathway

作　　者：谢晶 陆凌怡 XIE Jing;LU Lingyi(Department of Dermatology,Haishu District People's Hospital of Ningbo,Ningbo 315010,China;Department of Dermatology,Ningbo No.1 Hospital,Ningbo 315000,China)

机构地区：[1]宁波市海曙区人民医院皮肤科,浙江宁波315010 [2]宁波市第一医院皮肤科,浙江宁波315000

出　　处：《中国药学杂志》2024年第14期1312-1319,共8页Chinese Pharmaceutical Journal

基　　金：浙江省医药卫生科技计划项目资助(2022PY022)。

摘　　要：目的 探究维胺酯治疗大鼠耳部痤疮表皮过度角化的具体机制。方法 将30只雄性Sprague Dawley大鼠随机分成对照组、模型组和维胺酯治疗组。利用痤疮丙酸杆菌(Propionibacterium acnes,P.acnes)联合皮脂涂抹方法诱导大鼠耳部痤疮形成,建模成功后,通过灌胃给药,每12 h 1次,持续30 d。给药结束后,取大鼠痤疮组织,免疫组化,苏木精-伊红染色法检测组织病理,转录组测序分析结合分子对接预测维胺酯作用靶标,最后通过免疫印迹验证维胺酯对角化蛋白的影响。随后通过培养人角质形成细胞HaCat,考察维胺酯对热灭活后的P.acnes诱导HaCat细胞增殖及角化水平的影响,并通过沉默视黄醇结合蛋白(retinol binding protein 4,RBP4)验证维胺酯与RBP4的靶向关系。结果 维胺酯治疗30 d后,可显著改善大鼠耳部红肿、表皮增厚、炎症反应等症状,并抑制角化相关蛋白波形蛋白(vimentin)和角蛋白(keratin 6,KRT6)表达。转录组分析结果显示,维胺酯对细胞角化生物途径具有显著调控作用。分子对接结果显示RBP4可能是维胺酯发挥作用的关键靶点。免疫印迹结果证实维胺酯能够上调痤疮大鼠RBP4/视黄酸受体-γ(retinoic acid receptor γ,RARγ)蛋白表达。细胞层面,维胺酯可显著抑制P.acnes诱导后HaCat细胞的异常增殖及角化蛋白vimentin和KRT6的表达,并下调丝氨酸/苏氨酸蛋白激酶B(protein kinase B,也被称为Akt)蛋白磷酸化;而在转染RBP4 siRNA后,维胺酯对上述现象的调控作用均被显著逆转。结论 维胺酯通过激活RBP4/RARγ从而下调Akt蛋白磷酸化来抑制大鼠耳部表皮的过度角化来改善痤疮,并抑制由P.acnes诱导的人表皮角质形成细胞增殖及角化。OBJECTIVE To investigate the mechanism by which Viaminate treats hyperkeratosis of ear acne epidermis in rats.METHODS Thirty male SD rats were randomly divided into three groups:control group,model group,and viaminate-treated group.The formation of ear acne was induced by combining Propionibacterium acnes with a sebum smear.After successful modeling,viaminate was administered orally every 12 h for 30 d.Acne tissues were collected from the rats after the treatment,and immunohistochemical,Masson,and hematoxylin-eosin staining were performed to assess histopathology.Transcriptomic sequencing and molecular docking were conducted to analyze the target of viaminate.The effect of viaminate on the selected protein was confirmed using western blotting.Retinol binding protein 4(RBP4) was silenced in HaCat cells to verify the targeting relationship between viaminate and RBP4.RESULTS After 30 days of treatment with viaminate,significant improvements were observed in the symptoms of rat ear swelling,epidermal thickening,and inflammation.Additionally,the expression of the keratinization-related proteins,vimentin and keratin 6(KRT6),was inhibited.Transcriptomic analysis revealed that viaminate had a notable regulatory effect on the keratinization pathway.Molecular docking results suggested that RBP4 may be a critical target of viaminate.Western blotting results demonstrated that viaminate could upregulate the expression of RBP4/retinoic acid receptor γ(RARγ) protein in rats with acne.In addition,viaminate significantly inhibits abnormal proliferation and the expression of vimentin and KRT6 in HaCat cells induced by P.acnes.It also downregulates the phosphorylation of Akt.However,after transfection with RBP4 siRNA,the regulatory effects of viaminate on the aforementioned phenomena are significantly reversed.CONCLUSION Viaminate improves acne by reducing excessive keratinization in rat epidermis via activation of RBP4/RARγ and downregulation of Akt phosphorylation.It also inhibits the proliferation and keratinization of human

关 键 词：维胺酯 痤疮 角化过度 视黄醇结合蛋白 视黄酸受体-γ 

分 类 号：R965[医药卫生—药理学]