Hypertrophic chondrocytes(HCs)could transform into osteoblastic lineage cells while the pathophysiological implications of HC transformation remain largely unknown.Here,we generated a mouse line utilizing Col10a1-Cre ...Hypertrophic chondrocytes(HCs)could transform into osteoblastic lineage cells while the pathophysiological implications of HC transformation remain largely unknown.Here,we generated a mouse line utilizing Col10a1-Cre to induce DTA expression to genetically ablate HCs and their descendants.Col10a1-Cre;R26^(DTA/+)mice displayed dwarf phenotype,abnormal spongy bone,and significantly delayed drill-hole injuries healing,suggesting an indispensable role of HC lineage extension in bone growth and injury repair.Intriguingly,single-cell RNA sequencing analysis revealed the most significant loss of a cell cluster expressing multiple angiogenic factors(Pro-Angiogenic Descendants of HCs,PADs)among cells derived from Col10a1-Cre;R26^(DTA/+)and control femurs.In silico analysis of cell-cell communication supported Thrombospondin 4(THBS4)as a specific angiogenic factor mediating the crosstalk between PADs and vascular endothelial cells.Concordantly,analyses using immunostaining combined with tissue clearing revealed that PADs physically contacted with endothelial cells,whereas Col10a1-Cre;R26^(DTA/+)mice showed defective metaphyseal and cortical vessel formation and post-injury angiogenesis along with a significant loss of THBS4.Moreover,in vitro assays showed that supplying THBS4 was sufficient to promote proliferation and tube formation of endothelial cells and rescue defective angiogenesis of Col10a1-Cre;R26D TA/+metatarsal explants.Collectively,these findings demonstrate a critical role of PADs in bone growth and injury repair by secreting THBS4 to regulate angiogenesis.展开更多
目的:观察解毒化瘀健脾方对胃黏膜异型增生模型大鼠血小板反应蛋白1基因(thrombospondin 1,Thbs1)甲基化状态的影响,并探讨解毒化瘀健脾方治疗胃黏膜异型增生的可能机制.方法:将实验性胃黏膜异型增生病变模型大鼠分为:模型对照组(model ...目的:观察解毒化瘀健脾方对胃黏膜异型增生模型大鼠血小板反应蛋白1基因(thrombospondin 1,Thbs1)甲基化状态的影响,并探讨解毒化瘀健脾方治疗胃黏膜异型增生的可能机制.方法:将实验性胃黏膜异型增生病变模型大鼠分为:模型对照组(model control group,M G)、阳性对照组(positive control group,PCG)-西药维甲酸治疗组、解毒化瘀健脾方治疗组(Jiedu Huayu Jianpi Fang treatment group,A),并以健康大鼠为对照组(control group,CG),进行相应的药物干预;取胃黏膜组织,应用甲基化特异PCR技术检测Thbs1基因甲基化状态;HE染色观察各处理组胃黏膜组织结构变化差异.结果:CG组10只健康大鼠胃黏膜经检测Thbs1基因均未发生甲基化,胃黏膜异型增生MG组大鼠T h b s1基因的甲基化阳性检出率为33.33%(6/18);PCG组-西药维甲酸组同正常大鼠相同,Thbs1基因甲基化检出率为0.00%;A组治疗大鼠Thbs1基因甲基化比率(20.00%),相比MG组显著降低(P=0.0198).HE染色结果显示MG、PCG、A组呈现中轻度胃黏膜异型增生症状,经治疗后PCG、A组异型增生趋于正常.结论:解毒化瘀健脾方对发生异型增生的胃黏膜组织Thbs1基因具有显著地去甲基化作用.解毒化瘀健脾方治疗胃黏膜异型增生可能与该药物使Thbs1基因甲基化程度显著降低有关.展开更多
基金financial support from the National Natural Science Foundation of China(grants 82394442,82422043,82272442)the Key Industrial Chain Program of Shaanxi,China(No.2022ZDLSF02-12)。
文摘Hypertrophic chondrocytes(HCs)could transform into osteoblastic lineage cells while the pathophysiological implications of HC transformation remain largely unknown.Here,we generated a mouse line utilizing Col10a1-Cre to induce DTA expression to genetically ablate HCs and their descendants.Col10a1-Cre;R26^(DTA/+)mice displayed dwarf phenotype,abnormal spongy bone,and significantly delayed drill-hole injuries healing,suggesting an indispensable role of HC lineage extension in bone growth and injury repair.Intriguingly,single-cell RNA sequencing analysis revealed the most significant loss of a cell cluster expressing multiple angiogenic factors(Pro-Angiogenic Descendants of HCs,PADs)among cells derived from Col10a1-Cre;R26^(DTA/+)and control femurs.In silico analysis of cell-cell communication supported Thrombospondin 4(THBS4)as a specific angiogenic factor mediating the crosstalk between PADs and vascular endothelial cells.Concordantly,analyses using immunostaining combined with tissue clearing revealed that PADs physically contacted with endothelial cells,whereas Col10a1-Cre;R26^(DTA/+)mice showed defective metaphyseal and cortical vessel formation and post-injury angiogenesis along with a significant loss of THBS4.Moreover,in vitro assays showed that supplying THBS4 was sufficient to promote proliferation and tube formation of endothelial cells and rescue defective angiogenesis of Col10a1-Cre;R26D TA/+metatarsal explants.Collectively,these findings demonstrate a critical role of PADs in bone growth and injury repair by secreting THBS4 to regulate angiogenesis.
文摘目的:观察解毒化瘀健脾方对胃黏膜异型增生模型大鼠血小板反应蛋白1基因(thrombospondin 1,Thbs1)甲基化状态的影响,并探讨解毒化瘀健脾方治疗胃黏膜异型增生的可能机制.方法:将实验性胃黏膜异型增生病变模型大鼠分为:模型对照组(model control group,M G)、阳性对照组(positive control group,PCG)-西药维甲酸治疗组、解毒化瘀健脾方治疗组(Jiedu Huayu Jianpi Fang treatment group,A),并以健康大鼠为对照组(control group,CG),进行相应的药物干预;取胃黏膜组织,应用甲基化特异PCR技术检测Thbs1基因甲基化状态;HE染色观察各处理组胃黏膜组织结构变化差异.结果:CG组10只健康大鼠胃黏膜经检测Thbs1基因均未发生甲基化,胃黏膜异型增生MG组大鼠T h b s1基因的甲基化阳性检出率为33.33%(6/18);PCG组-西药维甲酸组同正常大鼠相同,Thbs1基因甲基化检出率为0.00%;A组治疗大鼠Thbs1基因甲基化比率(20.00%),相比MG组显著降低(P=0.0198).HE染色结果显示MG、PCG、A组呈现中轻度胃黏膜异型增生症状,经治疗后PCG、A组异型增生趋于正常.结论:解毒化瘀健脾方对发生异型增生的胃黏膜组织Thbs1基因具有显著地去甲基化作用.解毒化瘀健脾方治疗胃黏膜异型增生可能与该药物使Thbs1基因甲基化程度显著降低有关.
