Objective:To assess the safety and topical efficacy of prim-O-glucosylcimifugin(POG)and investigate the molecular mechanisms of its therapeutic effects in atopic dermatitis(AD).Methods:The effects of POG on human kera...Objective:To assess the safety and topical efficacy of prim-O-glucosylcimifugin(POG)and investigate the molecular mechanisms of its therapeutic effects in atopic dermatitis(AD).Methods:The effects of POG on human keratinocyte cell viability and its anti-inflammatory properties were evaluated using cell counting kit-8 assay and reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Subsequently,the impact of POG on the differentiation of cluster of differentiation(CD)4~+T cell subsets,including T-helper type(Th)1,Th2,Th17,and regulatory T(Treg),was examined through in vitro experiments.Network pharmacology analysis was used to elucidate POG's therapeutic mechanisms.Furthermore,the therapeutic potential of topically applied POG was further evaluated in a calcipotriol-induced mouse model of AD.The protein and transcript levels of inflammatory markers,including cytokines,lymphocyte-specific protein tyrosine kinase(Lck)mRNA,and LCK phosphorylation(p-LCK),were quantified using immunohistochemistry,RT-qPCR,and Western blot analysis.Results:POG was able to suppress cell proliferation and downregulate the transcription of interleukin 4(Il4)and Il13 mRNA.In vitro experiments indicated that POG significantly inhibited the differentiation of Th2 cells,whereas it exerted negligible influence on the differentiation of Th1,Th17 and Treg cells.Network pharmacology identified LCK as a key therapeutic target of POG.Moreover,the topical application of POG effectively alleviated skin lesions in the calcipotriol-induced AD mouse models without causing pathological changes in the liver,kidney or spleen tissues.POG significantly reduced the levels of Il4,Il5,Il13,and thymic stromal lymphopoietin(Tslp)m RNA in the AD mice.Concurrently,POG enhanced the expression of p-LCK protein and Lck mRNA.Conclusion:Our research revealed that POG inhibits Th2 cell differentiation by promoting p-LCK protein expression and hence effectively alleviates AD-related skin inflammation.展开更多
基金supported by the National Natural Science Foundation of China(No.82004359)Youth Talent Promotion Project of China Association of Traditional Chinese Medicine(2024–2026)Category B(No.2024-QNRC2-B04)+9 种基金Youth Medical Talents-Specialist Program of Shanghai“Rising Stars of Medical Talents”Youth Development ProgramHealth Young Talents of Shanghai Municipal Health Commission(No.2022YQ026)Shanghai Dermatology Research Center(No.2023ZZ02017)Shanghai Skin Disease Hospital demonstration research ward project(No.SHDC2023CRW009)Shanghai Key Discipline Construction Project of Traditional Chinese Medicine(No.shzyyzdxk-2024104)Shanghai Municipal Health Commission Health Industry Clinical Research Special Project(No.20224Y0373No.20234Y0075)Clinical Incubation Program(No.lcfy2023-08)Evidence-based dermatology base sponsored by State Administration of Traditional Chinese MedicineHigh-level Chinese Medicine Key Discipline Construction Project(Integrative Chinese and Western Medicine Clinic)of National Administration of TCM(No.zyyzdxk-2023065)。
文摘Objective:To assess the safety and topical efficacy of prim-O-glucosylcimifugin(POG)and investigate the molecular mechanisms of its therapeutic effects in atopic dermatitis(AD).Methods:The effects of POG on human keratinocyte cell viability and its anti-inflammatory properties were evaluated using cell counting kit-8 assay and reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Subsequently,the impact of POG on the differentiation of cluster of differentiation(CD)4~+T cell subsets,including T-helper type(Th)1,Th2,Th17,and regulatory T(Treg),was examined through in vitro experiments.Network pharmacology analysis was used to elucidate POG's therapeutic mechanisms.Furthermore,the therapeutic potential of topically applied POG was further evaluated in a calcipotriol-induced mouse model of AD.The protein and transcript levels of inflammatory markers,including cytokines,lymphocyte-specific protein tyrosine kinase(Lck)mRNA,and LCK phosphorylation(p-LCK),were quantified using immunohistochemistry,RT-qPCR,and Western blot analysis.Results:POG was able to suppress cell proliferation and downregulate the transcription of interleukin 4(Il4)and Il13 mRNA.In vitro experiments indicated that POG significantly inhibited the differentiation of Th2 cells,whereas it exerted negligible influence on the differentiation of Th1,Th17 and Treg cells.Network pharmacology identified LCK as a key therapeutic target of POG.Moreover,the topical application of POG effectively alleviated skin lesions in the calcipotriol-induced AD mouse models without causing pathological changes in the liver,kidney or spleen tissues.POG significantly reduced the levels of Il4,Il5,Il13,and thymic stromal lymphopoietin(Tslp)m RNA in the AD mice.Concurrently,POG enhanced the expression of p-LCK protein and Lck mRNA.Conclusion:Our research revealed that POG inhibits Th2 cell differentiation by promoting p-LCK protein expression and hence effectively alleviates AD-related skin inflammation.
