Ulcerative colitis(UC)is a chronic and non-specific inflammatory bowel disease(IBD).Huanglian Ganjiang decoction(HGD),derived from ancient book Beiji Qianjin Yao Fang,has demonstrated efficacy in treating UC patients ...Ulcerative colitis(UC)is a chronic and non-specific inflammatory bowel disease(IBD).Huanglian Ganjiang decoction(HGD),derived from ancient book Beiji Qianjin Yao Fang,has demonstrated efficacy in treating UC patients traditionally.Previous research established that the compatibility of cold herb Coptidis Rhizoma+Phellodendri Chinensis Cortex(CP)and hot herb Angelicae Sinensis Radix+Zingiberis Rhizoma(AZ)in HGD synergistically improved colitis mice.This study investigated the compatibility mechanisms through which CP and AZ regulated inflammatory balance in colitis mice.The experimental colitis model was established by administering 3%dextran sulphate sodium(DSS)to mice for 7 days,followed by CP,AZ and CPAZ treatment for an additional 7 days.M1/M2 macrophage polarization levels,glucose metabolites levels and pyruvate dehydrogenase kinase 4(PDK4)expression were analyzed using flow cytometry,Western blot,immunofluorescence and targeted glucose metabolomics.The findings indicated that CP inhibited M1 macrophage polarization,decreased inflammatory metabolites associated with tricarboxylic acid(TCA)cycle,and suppressed PDK4 expression and pyruvate dehydrogenase(PDH)(Ser-293)phosphorylation level.AZ enhanced M2 macrophage polarization,increased lactate axis metabolite lactate levels,and upregulated PDK4 expression and PDH(Ser-293)phosphorylation level.TCA cycle blocker AG-221 and adeno-associated virus(AAV)-PDK4 partially negated CP’s inhibition of M1 macrophage polarization.Lactate axis antagonist oxamate and PDK4 inhibitor dichloroacetate(DCA)partially reduced AZ’s activation of M2 macrophage polarization.In conclusion,the compatibility of CP and AZ synergistically alleviated colitis in mice through M1/M2 macrophage polarization balance via PDK4-mediated glucose metabolism reprogramming.Specifically,CP reduced M1 macrophage polarization by restoration of TCA cycle via PDK4 inhibition,while AZ increased M2 macrophage polarization through activation of PDK4/lactate axis.展开更多
目的:观察淫羊藿苷(ICA)对脂蛋白多糖诱导的巨噬细胞RAW264.7细胞M1/M2炎症表型转化作用。方法:采用M1/M2经典刺激方法后,Real-time PCR法检测M1型巨噬细胞标志基因IL-6、TNF-α和M2型巨噬细胞标志基因IL-10、Arg等的m RNA表达情况。并...目的:观察淫羊藿苷(ICA)对脂蛋白多糖诱导的巨噬细胞RAW264.7细胞M1/M2炎症表型转化作用。方法:采用M1/M2经典刺激方法后,Real-time PCR法检测M1型巨噬细胞标志基因IL-6、TNF-α和M2型巨噬细胞标志基因IL-10、Arg等的m RNA表达情况。并观察ICA不同浓度(50μg/m L、100μg/m L)以及以不同时间(6h、12h、24h)分别干预巨噬细胞M1及M2相关基因的表达变化情况。在将巨噬细胞诱导M1型后,通过Real-time PCR法检测ICA在最适浓度和最适时间干预下,M1及M2标志基因表达变化,并分析ICA对M1型巨噬细胞向M2亚型转化的诱导作用。结果:M1/M2经典方法刺激后,M1型巨噬细胞标志分子IL-6、TNF-αm RNA与M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05)。且ICA浓度为100μg/m L时,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),诱导巨噬细胞向M2方向分化。100μg/m L ICA干预12h后,对M1/M2不同标志分子m RNA表达变化产生明显影响(P<0.05)。100μg/m L ICA作用于LPS+IFN-γ诱导的M1巨噬细胞后,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),M1型巨噬细胞标志分子IL-6 m RNA表达下降(P<0.05)。结论:ICA可以诱导体外培养巨噬细胞RAW264.7向抗炎亚型M型方向分化,并可以诱导已经分化的促炎亚型M1巨噬细胞向抗炎亚型M2方向转化。展开更多
基金supported by the National Natural Science Foundation of China(Nos.82374325 and 82074322)GDAS'ProjectofScience and Technology Development(No.2022GDASZH-2022010110).
文摘Ulcerative colitis(UC)is a chronic and non-specific inflammatory bowel disease(IBD).Huanglian Ganjiang decoction(HGD),derived from ancient book Beiji Qianjin Yao Fang,has demonstrated efficacy in treating UC patients traditionally.Previous research established that the compatibility of cold herb Coptidis Rhizoma+Phellodendri Chinensis Cortex(CP)and hot herb Angelicae Sinensis Radix+Zingiberis Rhizoma(AZ)in HGD synergistically improved colitis mice.This study investigated the compatibility mechanisms through which CP and AZ regulated inflammatory balance in colitis mice.The experimental colitis model was established by administering 3%dextran sulphate sodium(DSS)to mice for 7 days,followed by CP,AZ and CPAZ treatment for an additional 7 days.M1/M2 macrophage polarization levels,glucose metabolites levels and pyruvate dehydrogenase kinase 4(PDK4)expression were analyzed using flow cytometry,Western blot,immunofluorescence and targeted glucose metabolomics.The findings indicated that CP inhibited M1 macrophage polarization,decreased inflammatory metabolites associated with tricarboxylic acid(TCA)cycle,and suppressed PDK4 expression and pyruvate dehydrogenase(PDH)(Ser-293)phosphorylation level.AZ enhanced M2 macrophage polarization,increased lactate axis metabolite lactate levels,and upregulated PDK4 expression and PDH(Ser-293)phosphorylation level.TCA cycle blocker AG-221 and adeno-associated virus(AAV)-PDK4 partially negated CP’s inhibition of M1 macrophage polarization.Lactate axis antagonist oxamate and PDK4 inhibitor dichloroacetate(DCA)partially reduced AZ’s activation of M2 macrophage polarization.In conclusion,the compatibility of CP and AZ synergistically alleviated colitis in mice through M1/M2 macrophage polarization balance via PDK4-mediated glucose metabolism reprogramming.Specifically,CP reduced M1 macrophage polarization by restoration of TCA cycle via PDK4 inhibition,while AZ increased M2 macrophage polarization through activation of PDK4/lactate axis.
文摘目的:观察淫羊藿苷(ICA)对脂蛋白多糖诱导的巨噬细胞RAW264.7细胞M1/M2炎症表型转化作用。方法:采用M1/M2经典刺激方法后,Real-time PCR法检测M1型巨噬细胞标志基因IL-6、TNF-α和M2型巨噬细胞标志基因IL-10、Arg等的m RNA表达情况。并观察ICA不同浓度(50μg/m L、100μg/m L)以及以不同时间(6h、12h、24h)分别干预巨噬细胞M1及M2相关基因的表达变化情况。在将巨噬细胞诱导M1型后,通过Real-time PCR法检测ICA在最适浓度和最适时间干预下,M1及M2标志基因表达变化,并分析ICA对M1型巨噬细胞向M2亚型转化的诱导作用。结果:M1/M2经典方法刺激后,M1型巨噬细胞标志分子IL-6、TNF-αm RNA与M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05)。且ICA浓度为100μg/m L时,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),诱导巨噬细胞向M2方向分化。100μg/m L ICA干预12h后,对M1/M2不同标志分子m RNA表达变化产生明显影响(P<0.05)。100μg/m L ICA作用于LPS+IFN-γ诱导的M1巨噬细胞后,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),M1型巨噬细胞标志分子IL-6 m RNA表达下降(P<0.05)。结论:ICA可以诱导体外培养巨噬细胞RAW264.7向抗炎亚型M型方向分化,并可以诱导已经分化的促炎亚型M1巨噬细胞向抗炎亚型M2方向转化。
