Article(id=1218297482681959277, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218297478793843630, articleNumber=1001-2494(2024)14-1312-08, orderNo=null, doi=10.11669/cpj.2024.14.007, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1709654400000, receivedDateStr=2024-03-06, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768394354744, onlineDateStr=2026-01-14, pubDate=1721577600000, pubDateStr=2024-07-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768394354744, onlineIssueDateStr=2026-01-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768394354744, creator=13701087609, updateTime=1768394354744, updator=13701087609, issue=Issue{id=1218297478793843630, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='14', pageStart='1273', pageEnd='1358', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768394353817, creator=13701087609, updateTime=1768394585064, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1218298448764387533, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218297478793843630, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1218298448764387534, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218297478793843630, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1312, endPage=1319, ext={EN=ArticleExt(id=1218297482921034607, articleId=1218297482681959277, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Viaminate Alleviates Hyperkeratosis of Ear Acne Epidermis in Rats by Activating RBP4/RARγ Pathway, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

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 epidermal keratinocytes induced by P. acnes in vitro.

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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。

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谢晶,女,本科,主治医师 研究方向:皮肤病临床及转化研究 Tel:(0574)87559632

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DOI: 10.1186/s40246-019-0248-9., articleTitle=Genetics and functions of the retinoic acid pathway, with special emphasis on the eye, refAbstract=null), Reference(id=1218297491448053858, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, doi=null, pmid=null, pmcid=null, year=1994, volume=269, issue=32, pageStart=20629, pageEnd=20635, url=null, language=null, rfNumber=[15], rfOrder=14, authorNames=FISHER G J, TALWAR H S, XIAO J H, journalName=J Biol Chem, refType=null, unstructuredReference=FISHER G J, TALWAR H S, XIAO J H, et al. Immunological identification and functional quantitation of retinoic acid and retinoid X receptor proteins in human skin[J]. J Biol Chem, 1994, 269(32): 20629-20635., articleTitle=Immunological identification and functional quantitation of retinoic acid and retinoid X receptor proteins in human skin, refAbstract=null), Reference(id=1218297491540328548, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, doi=null, pmid=null, pmcid=null, year=2018, volume=28, issue=10, pageStart=1736, pageEnd=1741, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=THOREAU E, ARLABOSSE J M, BOUIX-PETER C, journalName=Bioorg Med Chem Lett, refType=null, unstructuredReference=THOREAU E, ARLABOSSE J M, BOUIX-PETER C, et al. Structure-based design of Trifarotene (CD5789), a potent and selective RARγ agonist for the treatment of acne[J]. 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DOI: 10.1038/s41419-022-05284-9., articleTitle=Retinoic acid and RARγ maintain satellite cell quiescence through regulation of translation initiation, refAbstract=null), Reference(id=1218297491737460845, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, doi=null, pmid=null, pmcid=null, year=2021, volume=10, issue=5, pageStart=1219, pageEnd=null, url=null, language=null, rfNumber=[18], rfOrder=17, authorNames=TENG Y, FAN Y, MA J, journalName=Cells, refType=null, unstructuredReference=TENG Y, FAN Y, MA J, et al. The PI3K/Akt Pathway: Emerging roles in skin homeostasis and a group of non-malignant skin disorders[J]. Cells, 2021, 10(5): 1219. DOI: 10.3390/cells10051219., articleTitle=The PI3K/Akt Pathway: Emerging roles in skin homeostasis and a group of non-malignant skin disorders, refAbstract=null), Reference(id=1218297491808764015, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, doi=null, pmid=null, pmcid=null, year=2011, volume=30, issue=4, pageStart=457, pageEnd=470, url=null, language=null, rfNumber=[19], rfOrder=18, authorNames=ZHU Q S, ROSENBLATT K, HUANG K L, journalName=Oncogene, refType=null, unstructuredReference=ZHU Q S, ROSENBLATT K, HUANG K L, et al. Vimentin is a novel AKT1 target mediating motility and invasion[J]. Oncogene, 2011, 30(4): 457-470., articleTitle=Vimentin is a novel AKT1 target mediating motility and invasion, refAbstract=null), Reference(id=1218297491892650098, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, doi=null, pmid=null, pmcid=null, year=2017, volume=331, issue=null, pageStart=41, pageEnd=53, url=null, language=null, rfNumber=[20], rfOrder=19, authorNames=SLUSSER-NORE A, GARRETT S H, ZHOU X D, journalName=Toxicol Appl Pharmacol, refType=null, unstructuredReference=SLUSSER-NORE A, GARRETT S H, ZHOU X D, et al. The expression of keratin 6 is regulated by the activation of the ERK1/2 pathway in arsenite transformed human urothelial cells[J]. Toxicol Appl Pharmacol, 2017, 331: 41-53., articleTitle=The expression of keratin 6 is regulated by the activation of the ERK1/2 pathway in arsenite transformed human urothelial cells, refAbstract=null)], funds=[Fund(id=1218297488918888495, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, awardId=2022PY022, language=CN, fundingSource=浙江省医药卫生科技计划项目(2022PY022), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1218297484607144847, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, xref=1, ext=[AuthorCompanyExt(id=1218297484615533456, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, companyId=1218297484607144847, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=¹ Department of Dermatology, Haishu District People's Hospital of Ningbo, Ningbo 315010, China), AuthorCompanyExt(id=1218297484619727761, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, companyId=1218297484607144847, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=¹ 宁波市海曙区人民医院皮肤科, 浙江 宁波 315010)]), AuthorCompany(id=1218297484686836628, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, xref=2, ext=[AuthorCompanyExt(id=1218297484695225237, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, companyId=1218297484686836628, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=² Department of Dermatology, Ningbo No.1 Hospital, Ningbo 315000, China), AuthorCompanyExt(id=1218297484703613846, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, companyId=1218297484686836628, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=² 宁波市第一医院皮肤科, 浙江 宁波 315000)])], figs=[ArticleFig(id=1218297487677375481, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.1, caption=Effect of viaminate on progression of ear acne induced by Propionibacterium acnes in rats

A-appearance of rat left ear; B-HE staining(scale bar=200 μm)

, figureFileSmall=hdAqk6NP0S3NXIOES8aZNg==, figureFileBig=mucH2CheRB9vx2abYgeJmQ==, tableContent=null), ArticleFig(id=1218297487782232065, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图1, caption=维胺酯对大鼠耳部痤疮模型外观和皮肤病理影响

A-大鼠左耳外观;B-苏木精和伊红染色(标尺=200 μm)

, figureFileSmall=hdAqk6NP0S3NXIOES8aZNg==, figureFileBig=mucH2CheRB9vx2abYgeJmQ==, tableContent=null), ArticleFig(id=1218297488000335885, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.2, caption=Effects of viaminate treatment on the expression and distribution of keratinization related proteins KRT6 and vimentin. n=3,$\bar{x}±s$

A-immunohistochemical results, and IPP6.0 was used to quantify the positive intensity; B-results of Western blot, and the optical density values of protein bands were quantified by IPP6.0; ¹⁾P<0.05, compared with the control group;²⁾P<0.05, compared with model group.

, figureFileSmall=JBRC8spjNcS6oKAnvzKTbQ==, figureFileBig=gxIPaQBBySD7H2Vr6109wg==, tableContent=null), ArticleFig(id=1218297488088416271, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图2, caption=维胺酯处理后对角化相关蛋白KRT6和vimentin表达及分布影响。n=3,$\bar{x}±s$

A-免疫组化结果,IPP6.0定量阳性强度;B-Western blot结果和IPP6.0定量蛋白条带的光密度值;与对照组比较,¹⁾P<0.05;与模型组比较,²⁾P<0.05。

, figureFileSmall=JBRC8spjNcS6oKAnvzKTbQ==, figureFileBig=gxIPaQBBySD7H2Vr6109wg==, tableContent=null), ArticleFig(id=1218297488189079574, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.3, caption=Transcriptome analysis of key regulatory genes and pathways of viaminate in the treatment of acne rats. n=3,$\bar{x}±s$

A-analysis results of KEGG enrichment in control and model groups; B-the results of KEGG enrichment in the treatment group and model group; C-possible targets of viaminate screening by Venn diagram; D-transcriptome analysis of RXRG, RARG, RBP4, and RARB gene expression levels.

, figureFileSmall=Tq2ApkntU+wPL3i2rljFBg==, figureFileBig=7Y/VyUMt6n0xCkDPF/QHSg==, tableContent=null), ArticleFig(id=1218297488277159961, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图3, caption=转录组分析维胺酯对痤疮大鼠治疗的关键调控基因及途径。n=3,$\bar{x}±s$

A-对照组和模型组KEGG富集分析结果;B-治疗组和模型组KEGG富集结果;C-维胺酯筛选的可能靶点;D-RXRG, RARG, RBP4和RARB基因表达水平的转录组分析。

, figureFileSmall=Tq2ApkntU+wPL3i2rljFBg==, figureFileBig=7Y/VyUMt6n0xCkDPF/QHSg==, tableContent=null), ArticleFig(id=1218297488356851739, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.4, caption=Effect of viaminate on the expression of RBP4/RARγ proteins in acne rats. n=3,$\bar{x}±s$

A-3D structure simulation docking results of viaminate and RBP4 protein; B-E-western blot was used to detect the expression level of RBP4/RARγ and Akt protein. ¹⁾P<0.05, compared with the control group; ²⁾P<0.05, compared with model group, the difference is statistically significant.

, figureFileSmall=MLhsX/5teGFjRulfgd8RhA==, figureFileBig=BcNpsFOfY4FJFwkydowTRA==, tableContent=null), ArticleFig(id=1218297488453320734, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图4, caption=维胺酯对痤疮大鼠RBP4/RARγ蛋白表达的影响。n=3,$\bar{x}±s$

A-维胺酯与RBP4蛋白的三维结构模拟对接结果;B~E-Western blot检测RBP4/RARγ以及Akt蛋白表达水平;与对照组比较,¹⁾P<0.05;与模型组比较,²⁾P<0.05。

, figureFileSmall=MLhsX/5teGFjRulfgd8RhA==, figureFileBig=BcNpsFOfY4FJFwkydowTRA==, tableContent=null), ArticleFig(id=1218297488553984034, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.5, caption=Effects of interfering RBP4 on viaminate treatment of HKP-induced abnormal proliferation of HaCat cells。n=3,$\bar{x}±s$

A-C-MTT assay was used to detect the effect of different doses of heat-inactivated P. acnes, viaminate, and combined treatment on the viability of HaCat cells. D-qRT-PCR was used to detect the interference efficiency of the RBP4 gene in HaCat cells transfected with RBP4 siRNA; E-The interference efficiency of RBP4 protein in HaCat cells transfected with RBP4 siRNA was verified by Western blot. F-MTT assay of the effect of RBP4 siRNA on changes in cell viability levels; ¹⁾P<0.05, compared with the control group; ²⁾P<0.05, compared with 400 μg·mL^-1 HKP group; ³⁾P<0.05, compared with viaminate group.

, figureFileSmall=Sq/tshDtTM7pr4ZZQ8LgOw==, figureFileBig=jzSHMLhfG/iNKzlnkP3w/Q==, tableContent=null), ArticleFig(id=1218297488654647333, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图5, caption=干扰视黄醇结合蛋白(retinol-binding protein 4,RBP4)对维胺酯治疗热灭活的痤疮丙酸杆菌(heat-killed P. acnes,HKP)诱导的HaCat细胞异常增殖的影响。n=3,$\bar{x}±s$

A~C-MTT法检测不同剂量热灭活P. acnes、维胺酯及联合处理对HaCat细胞活力的影响;D-实时荧光定量聚合酶链式反应检测RBP4 siRNA转染HaCat细胞后对RBP4基因的干扰效率;E-Western blot验证RBP4 siRNA对HaCat细胞RBP4蛋白的干扰效率;F-MTT法检测RBP4 siRNA对细胞活力水平的影响;与对照组相比,¹⁾P<0.05;与400 μg·mL^-1 HKP组比较,²⁾P<0.05; 与Via组相比,³⁾P<0.05。

, figureFileSmall=Sq/tshDtTM7pr4ZZQ8LgOw==, figureFileBig=jzSHMLhfG/iNKzlnkP3w/Q==, tableContent=null), ArticleFig(id=1218297488755310633, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=EN, label=Fig.6, caption=Viaminate regulates the expression of keratinizing proteins in HaCat cells induced by HKP through RBP4/RARγ/Akt pathway。n=3,$\bar{x}±s$

A-the expressions of RBP4/RARγ protein and phosphorylated Akt protein were detected by western blot, and the expressions of Akt downstream keratosis-related proteins Vimentin and KRT6 were detected; B-F-quantitative statistics of optical density values of protein bands detected by western blot; ¹⁾P<0.05, compared with the control group; ²⁾P<0.05, compared with 400 μg·mL^-1 HKP group; ³⁾P<0.05, compared with viaminate group.

, figureFileSmall=q8/+Pz9yjPbhOkgBYCYYUg==, figureFileBig=gH5O73LBpzFItl00kNeGyw==, tableContent=null), ArticleFig(id=1218297488830808107, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218297482681959277, language=CN, label=图6, caption=维胺酯通过RBP4/RARγ/Akt途径调控HKP诱导的HaCat细胞角化相关蛋白的表达。n=3,$\bar{x}±s$

A-采用Western blot检测RBP4/RARγ蛋白和磷酸化Akt蛋白表达,并检测Akt下游角化相关蛋白波形蛋白和角蛋白(keratin 6,KRT6)的表达;B~F-Western blot检测结果的统计;与对照组比较,¹⁾P<0.05;与400 μg·mL^-1 HKP组比较,²⁾P<0.05;与维胺酯(viaminate)组比较,³⁾P<0.05。

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