目的:探讨M2型肿瘤相关巨噬细胞(M2 type tumor-associated macrophage,M2-TAMs)介导白细胞介素6(interleukin-6,IL-6)-Janus激酶2(Janus kinase 2,JAK2)/信号转导及转录激活因子3(signal transducer and activator of transcription 3,...目的:探讨M2型肿瘤相关巨噬细胞(M2 type tumor-associated macrophage,M2-TAMs)介导白细胞介素6(interleukin-6,IL-6)-Janus激酶2(Janus kinase 2,JAK2)/信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)轴促进胃癌干性和侵袭迁移的作用机制。方法:应用TCGA数据库分析胃癌及癌旁正常组织M2-TAMs的分布与干性指标的关系,并在胃癌患者组织水平进行验证。体外通过成球实验检测胃癌细胞的干性潜能并构建M2-TAMs体外共培养体系,运用qRT-PCR和Western blot技术检测肿瘤干性标志物CD44和醛脱氢酶1(aldehyde dehydrogenase 1,ALDH1)的变化,通过Transwell实验探究胃癌迁移、侵袭能力的变化。利用转录组测序筛选差异基因,通过KEGG富集分析探寻关键通路,并采用中和抗体及通路抑制剂进行相关性验证。结果:TCGA数据库分析和体内组织验证结果均表明胃癌组织中M2-TAMs的密度显著高于癌旁正常组织,且其数量的增加与肿瘤干性标志(CD44和ALDH1)呈正相关。与M2-TAMs共培养的胃癌细胞在CD44和ALDH1的mRNA及蛋白表达水平较对照组显著升高,同时共培养组肿瘤细胞成球率和侵袭迁移能力均显著增强,JAK2/STAT3信号通路关键蛋白表达水平显著升高。KEGG通路分析发现STAT3是肿瘤干性调控的关键基因。与极化前的巨噬细胞相比,极化的M2-TAMs有22个上调分泌因子,其中IL-6显著升高,且与患者不良预后相关。加入stattic(选择性STAT3信号通路抑制剂)和(或)IL-6中和抗体均能抑制胃癌细胞的干性、迁移和侵袭。以上结果P值均<0.05。结论:M2-TAMs通过分泌IL-6介导JAK2/STAT3信号通路激活胃癌细胞干性潜能,进而促进胃癌的侵袭和迁移。展开更多
Objective:To investigate the effect of pectic polysaccharides isolated from Rauvolfia verticillata on ulcerative colitis and its underlying mechanisms.Methods:Pectic polysaccharides were characterized using high-perfo...Objective:To investigate the effect of pectic polysaccharides isolated from Rauvolfia verticillata on ulcerative colitis and its underlying mechanisms.Methods:Pectic polysaccharides were characterized using high-performance liquid chromatography with 1-phenyl-3-methyl-5-pyrazolone pre-column derivatization,phenol-sulfuric acid assay,and gel permeation chromatography.HT-29 cells were stimulated with lipopolysaccharide and then treated with pectic polysaccharides;conditioned medium was applied to THP-1-derived macrophages to assess cell viability and polarization,while tight junction protein expression was analyzed in HT-29 cells.Furthermore,a mouse model of dextran sulfate sodium-induced colitis was treated with oral pectic polysaccharides or NOS2 overexpression.Body weight,disease activity index,colon length,histopathology,and the protein expression related to the JAK2/STAT3-NOS2 signaling were evaluated.Results:The pectic polysaccharide was characterized as an acidic pectic polysaccharide,primarily composed of galacturonic acid and various neutral sugars,with a narrow molecular weight distribution and high purity.Pectic polysaccharides significantly enhanced THP-1 macrophage viability,promoted M1 to M2 polarization,and upregulated the expression of epithelial tight junction proteins.In addition,pectic polysaccharide treatment attenuated body weight loss,lowered disease activity index scores and improved colon histology in mice with dextran sulfate sodium-induced colitis.It also reduced JAK2/STAT3 phosphorylation and NOS2 expression,and increased the expression of tight junction proteins(ZO-1,occludin,and claudin-1).Conclusions:Pectic polysaccharides attenuate ulcerative colitis by increasing M2-related macrophage markers,inhibiting the JAK2/STAT3-NOS2 signaling,and enhancing epithelial barrier-related protein expression.These findings support pectic polysaccharides as a natural candidate for the treatment of ulcerative colitis.展开更多
文摘目的:探讨M2型肿瘤相关巨噬细胞(M2 type tumor-associated macrophage,M2-TAMs)介导白细胞介素6(interleukin-6,IL-6)-Janus激酶2(Janus kinase 2,JAK2)/信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)轴促进胃癌干性和侵袭迁移的作用机制。方法:应用TCGA数据库分析胃癌及癌旁正常组织M2-TAMs的分布与干性指标的关系,并在胃癌患者组织水平进行验证。体外通过成球实验检测胃癌细胞的干性潜能并构建M2-TAMs体外共培养体系,运用qRT-PCR和Western blot技术检测肿瘤干性标志物CD44和醛脱氢酶1(aldehyde dehydrogenase 1,ALDH1)的变化,通过Transwell实验探究胃癌迁移、侵袭能力的变化。利用转录组测序筛选差异基因,通过KEGG富集分析探寻关键通路,并采用中和抗体及通路抑制剂进行相关性验证。结果:TCGA数据库分析和体内组织验证结果均表明胃癌组织中M2-TAMs的密度显著高于癌旁正常组织,且其数量的增加与肿瘤干性标志(CD44和ALDH1)呈正相关。与M2-TAMs共培养的胃癌细胞在CD44和ALDH1的mRNA及蛋白表达水平较对照组显著升高,同时共培养组肿瘤细胞成球率和侵袭迁移能力均显著增强,JAK2/STAT3信号通路关键蛋白表达水平显著升高。KEGG通路分析发现STAT3是肿瘤干性调控的关键基因。与极化前的巨噬细胞相比,极化的M2-TAMs有22个上调分泌因子,其中IL-6显著升高,且与患者不良预后相关。加入stattic(选择性STAT3信号通路抑制剂)和(或)IL-6中和抗体均能抑制胃癌细胞的干性、迁移和侵袭。以上结果P值均<0.05。结论:M2-TAMs通过分泌IL-6介导JAK2/STAT3信号通路激活胃癌细胞干性潜能,进而促进胃癌的侵袭和迁移。
基金supported by the Key Research and Development Project of Hainan Province(ZDYF2022SHFZ099)the Academic Enhancement Support Program of Hainan Medical University(XSTS2025040 and XSTS2025063).
文摘Objective:To investigate the effect of pectic polysaccharides isolated from Rauvolfia verticillata on ulcerative colitis and its underlying mechanisms.Methods:Pectic polysaccharides were characterized using high-performance liquid chromatography with 1-phenyl-3-methyl-5-pyrazolone pre-column derivatization,phenol-sulfuric acid assay,and gel permeation chromatography.HT-29 cells were stimulated with lipopolysaccharide and then treated with pectic polysaccharides;conditioned medium was applied to THP-1-derived macrophages to assess cell viability and polarization,while tight junction protein expression was analyzed in HT-29 cells.Furthermore,a mouse model of dextran sulfate sodium-induced colitis was treated with oral pectic polysaccharides or NOS2 overexpression.Body weight,disease activity index,colon length,histopathology,and the protein expression related to the JAK2/STAT3-NOS2 signaling were evaluated.Results:The pectic polysaccharide was characterized as an acidic pectic polysaccharide,primarily composed of galacturonic acid and various neutral sugars,with a narrow molecular weight distribution and high purity.Pectic polysaccharides significantly enhanced THP-1 macrophage viability,promoted M1 to M2 polarization,and upregulated the expression of epithelial tight junction proteins.In addition,pectic polysaccharide treatment attenuated body weight loss,lowered disease activity index scores and improved colon histology in mice with dextran sulfate sodium-induced colitis.It also reduced JAK2/STAT3 phosphorylation and NOS2 expression,and increased the expression of tight junction proteins(ZO-1,occludin,and claudin-1).Conclusions:Pectic polysaccharides attenuate ulcerative colitis by increasing M2-related macrophage markers,inhibiting the JAK2/STAT3-NOS2 signaling,and enhancing epithelial barrier-related protein expression.These findings support pectic polysaccharides as a natural candidate for the treatment of ulcerative colitis.
