Diabetic retinopathy(DR),a common complication of diabetes,is characterized by retinal angiogenesis and inflammation.The role of hepatoma-derived growth factor(HDGF)in mediating inflammation during DR remains unclear....Diabetic retinopathy(DR),a common complication of diabetes,is characterized by retinal angiogenesis and inflammation.The role of hepatoma-derived growth factor(HDGF)in mediating inflammation during DR remains unclear.We measured HDGF levels in the aqueous humor and found that HDGF was increased in DR but decreased after anti-angiogenesis treatment.Using public single-cell RNA sequencing datasets,we found that elevated HDGF in DR was mainly produced by Müller cells and targeted microglia.Additionally,integrin beta 2(Itgb2),a target gene of HDGF that induces microglial activation,was significantly upregulated in DR.To verify these results,we performed enzyme-linked immunosorbent assays,quantitative reverse transcription-PCR,Western blotting,and fluorescence immunostaining in cultured Müller and microglial cells treated with HDGF or anti-HDGF,as well as in DR mice receiving intravitreal injections of HDGF or its antibody.Exogenous HDGF further promoted microglial activation,migration,and secretion of pro-inflammatory cytokines,while neutralization of HDGF suppressed these effects caused by high glucose.Furthermore,the HDGF receptor nucleolin was overexpressed in microglia under high glucose stimulation.Therefore,blocking HDGF from Müller cells in DR reduced the excessive inflammatory response in microglia,highlighting HDGF as a potential therapeutic target.展开更多
目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹...目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹祺胶囊0.4 g/kg给药组(BQ)的左膝关节组织,采用TMT定量蛋白质组学技术分析鉴定各组大鼠膝关节组织中的蛋白,以表达差异倍数≥1.2或差异倍数≤0.83且P<0.05筛选M vs C、BQ vs M、BQ vs C差异表达蛋白,并分析经痹祺胶囊干预后有回调趋势的差异表达蛋白。最后通过GeneCards、OMIN数据库检索RA疾病靶点,与回调蛋白进行交集分析得到与RA疾病相关的回调蛋白,对其进行基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,明晰痹祺胶囊对RA的作用机制。结果 经痹祺胶囊干预后有121个差异蛋白具有回调趋势,其中38个与RA疾病相关,如β2整合素(integrin subunit beta 2,ITGB2)、钙调蛋白2(calmodulin 2,CALM2)、钙调磷酸酶B亚基(calcineurin subunit B type 1,PPP3R1)、前列腺素合成酶(prostacyclin synthase,PTGIS)等。生物信息学分析发现这些回调蛋白参与了与RA相关的多条信号通路,如ITGB2参与调节RA及白细胞跨内皮细胞迁移信号通路;CALM2、PPP3R1通过调节钙信号通路,进而调节T细胞、B细胞受体信号通路、T细胞分化及破骨细胞分化等信号通路;PTGIS参与花生四烯酸代谢途径等。结论 痹祺胶囊可能通过干预ITGB2、PPP3R1、CALM2、PTGIS等蛋白的表达,调节了与RA相关的多条信号通路,发挥免疫抑制、抗炎、活血、抑制骨破坏和骨吸收、抑制血管翳生成等多方面药理作用,从而达到治疗RA的效果。展开更多
基金supported by National Natural Science Foundation of China(Grant No.81900873 to A.Q.)the Jiangsu Provincial Key Research and Development Programme-social development(Grant No.BE2023777 to W.Z.)+1 种基金the Key Medical Research Project of Jiangsu Commission of Health(Grant No.H2022185 to W.Z.)the Clinical Capacity Enhancement Project of Jiangsu Province Hospital(Grant No.JSPH-MB-2023-18 to W.Z.)。
文摘Diabetic retinopathy(DR),a common complication of diabetes,is characterized by retinal angiogenesis and inflammation.The role of hepatoma-derived growth factor(HDGF)in mediating inflammation during DR remains unclear.We measured HDGF levels in the aqueous humor and found that HDGF was increased in DR but decreased after anti-angiogenesis treatment.Using public single-cell RNA sequencing datasets,we found that elevated HDGF in DR was mainly produced by Müller cells and targeted microglia.Additionally,integrin beta 2(Itgb2),a target gene of HDGF that induces microglial activation,was significantly upregulated in DR.To verify these results,we performed enzyme-linked immunosorbent assays,quantitative reverse transcription-PCR,Western blotting,and fluorescence immunostaining in cultured Müller and microglial cells treated with HDGF or anti-HDGF,as well as in DR mice receiving intravitreal injections of HDGF or its antibody.Exogenous HDGF further promoted microglial activation,migration,and secretion of pro-inflammatory cytokines,while neutralization of HDGF suppressed these effects caused by high glucose.Furthermore,the HDGF receptor nucleolin was overexpressed in microglia under high glucose stimulation.Therefore,blocking HDGF from Müller cells in DR reduced the excessive inflammatory response in microglia,highlighting HDGF as a potential therapeutic target.
文摘目的 利用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎(rheumatoid arthritis,RA)的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型,并给予痹祺胶囊进行干预,取对照组(C)、模型组(M)、痹祺胶囊0.4 g/kg给药组(BQ)的左膝关节组织,采用TMT定量蛋白质组学技术分析鉴定各组大鼠膝关节组织中的蛋白,以表达差异倍数≥1.2或差异倍数≤0.83且P<0.05筛选M vs C、BQ vs M、BQ vs C差异表达蛋白,并分析经痹祺胶囊干预后有回调趋势的差异表达蛋白。最后通过GeneCards、OMIN数据库检索RA疾病靶点,与回调蛋白进行交集分析得到与RA疾病相关的回调蛋白,对其进行基因本体(gene ontology,GO)功能和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,明晰痹祺胶囊对RA的作用机制。结果 经痹祺胶囊干预后有121个差异蛋白具有回调趋势,其中38个与RA疾病相关,如β2整合素(integrin subunit beta 2,ITGB2)、钙调蛋白2(calmodulin 2,CALM2)、钙调磷酸酶B亚基(calcineurin subunit B type 1,PPP3R1)、前列腺素合成酶(prostacyclin synthase,PTGIS)等。生物信息学分析发现这些回调蛋白参与了与RA相关的多条信号通路,如ITGB2参与调节RA及白细胞跨内皮细胞迁移信号通路;CALM2、PPP3R1通过调节钙信号通路,进而调节T细胞、B细胞受体信号通路、T细胞分化及破骨细胞分化等信号通路;PTGIS参与花生四烯酸代谢途径等。结论 痹祺胶囊可能通过干预ITGB2、PPP3R1、CALM2、PTGIS等蛋白的表达,调节了与RA相关的多条信号通路,发挥免疫抑制、抗炎、活血、抑制骨破坏和骨吸收、抑制血管翳生成等多方面药理作用,从而达到治疗RA的效果。
