目的探讨炎症因子白介素-1β(interleukin-1β,IL-1β)对软骨表层细胞(articular cartilage superficial zone cells,ACSCs)衰老的调控作用和相关机制。方法用纤连蛋白黏附法分离培养ACSCs,根据实验分为对照组和IL-1β处理组,采用MTT检...目的探讨炎症因子白介素-1β(interleukin-1β,IL-1β)对软骨表层细胞(articular cartilage superficial zone cells,ACSCs)衰老的调控作用和相关机制。方法用纤连蛋白黏附法分离培养ACSCs,根据实验分为对照组和IL-1β处理组,采用MTT检测细胞增殖情况,衰老相关-β-半乳糖苷酶(SA-β-gal)染色检测SA-β-gal阳性细胞数,Western blot检测p21和p53蛋白水平,实时荧光定量PCR(RT-qPCR)检测衰老相关分泌表型(senescence-associated secretory phenotype,SASP)因子il-6、Mmp-3和Mmp-13 m RNA水平,CellROX试剂检测活性氧(reactive oxygen species,ROS)水平,JC-1试剂检测线粒体膜电位。最后,取ACSCs设IL-1β处理组和IL-1β+N-乙酰半胱氨酸(N-acetylcysteine,NAC)处理组,检测NAC处理后对IL-1β引起的细胞ROS水平、细胞增殖、SA-β-gal阳性细胞数、p21和p53蛋白水平和SASP分泌因子水平的影响。结果IL-1β处理导致ACSCs出现衰老相关表型,包括增殖减慢(P<0.05)、SA-β-gal染色阳性细胞增加(P<0.05)、p53和p21蛋白水平升高(P<0.05)、SASP因子il-6、Mmp-13和Mmp-3 m RNA水平升高(P<0.05)。进一步研究发现,IL-1β可以导致ACSCs中线粒体膜电位降低(P<0.05),ROS增加(P<0.05),而采用抗氧化剂NAC处理可以明显缓解IL-1β导致的ACSCs中ROS增加和ACSCs衰老相关表型(P<0.05)。结论IL-1β通过诱导氧化应激促进ACSCs衰老,这为从抑制ACSCs衰老和拮抗ACSCs氧化应激损伤角度治疗骨关节炎提供了实验依据。展开更多
We have investigated how gastric H. pylori infection affects antrum secretory cell types by studying the expression of secretory proteins in antrum epithelium. Antrum biopsy specimens were prospectively collected from...We have investigated how gastric H. pylori infection affects antrum secretory cell types by studying the expression of secretory proteins in antrum epithelium. Antrum biopsy specimens were prospectively collected from 102 individuals (49 H. pylori-infected). Immunohistochemistry was performed for secretory mucins (MUC5AC, MUC5B, MUC6), Trefoil factor family (TFF)-peptides (TFF1, TF-F2), endocrine peptides (gastrin, chromogranin A), and proliferating cells (Ki-67). Protein expression was quant-ified morphometrically. H. pylori infection was significantly correlated to mucosal inflammation and to epithelial atrophy and proliferation. In H. pylori-infected patients the number of proliferating cells increased significantly, and the zone of proliferating cells shifted toward the surface epithelium of the antral glands. Infection was correlated with decreased MUC5AC, TFF1, and TFF2 expression and increased MUC6 and MUC5B expression. Endocrine cells expressing chromagranin A and gastrin shifted toward the surface epithelium of the antral glands in H. pylori-infected patients. H. pylori infection and concomitant inflammation induced increased epithelial proliferation and triggered coordinate deregulation of secretory cell populations in the antrum. In particular, infection led to a coordinated increase in cells expressing MUC6 and MUC5B at the expense of MUC5AC-producing cells.展开更多
文摘目的探讨炎症因子白介素-1β(interleukin-1β,IL-1β)对软骨表层细胞(articular cartilage superficial zone cells,ACSCs)衰老的调控作用和相关机制。方法用纤连蛋白黏附法分离培养ACSCs,根据实验分为对照组和IL-1β处理组,采用MTT检测细胞增殖情况,衰老相关-β-半乳糖苷酶(SA-β-gal)染色检测SA-β-gal阳性细胞数,Western blot检测p21和p53蛋白水平,实时荧光定量PCR(RT-qPCR)检测衰老相关分泌表型(senescence-associated secretory phenotype,SASP)因子il-6、Mmp-3和Mmp-13 m RNA水平,CellROX试剂检测活性氧(reactive oxygen species,ROS)水平,JC-1试剂检测线粒体膜电位。最后,取ACSCs设IL-1β处理组和IL-1β+N-乙酰半胱氨酸(N-acetylcysteine,NAC)处理组,检测NAC处理后对IL-1β引起的细胞ROS水平、细胞增殖、SA-β-gal阳性细胞数、p21和p53蛋白水平和SASP分泌因子水平的影响。结果IL-1β处理导致ACSCs出现衰老相关表型,包括增殖减慢(P<0.05)、SA-β-gal染色阳性细胞增加(P<0.05)、p53和p21蛋白水平升高(P<0.05)、SASP因子il-6、Mmp-13和Mmp-3 m RNA水平升高(P<0.05)。进一步研究发现,IL-1β可以导致ACSCs中线粒体膜电位降低(P<0.05),ROS增加(P<0.05),而采用抗氧化剂NAC处理可以明显缓解IL-1β导致的ACSCs中ROS增加和ACSCs衰老相关表型(P<0.05)。结论IL-1β通过诱导氧化应激促进ACSCs衰老,这为从抑制ACSCs衰老和拮抗ACSCs氧化应激损伤角度治疗骨关节炎提供了实验依据。
文摘We have investigated how gastric H. pylori infection affects antrum secretory cell types by studying the expression of secretory proteins in antrum epithelium. Antrum biopsy specimens were prospectively collected from 102 individuals (49 H. pylori-infected). Immunohistochemistry was performed for secretory mucins (MUC5AC, MUC5B, MUC6), Trefoil factor family (TFF)-peptides (TFF1, TF-F2), endocrine peptides (gastrin, chromogranin A), and proliferating cells (Ki-67). Protein expression was quant-ified morphometrically. H. pylori infection was significantly correlated to mucosal inflammation and to epithelial atrophy and proliferation. In H. pylori-infected patients the number of proliferating cells increased significantly, and the zone of proliferating cells shifted toward the surface epithelium of the antral glands. Infection was correlated with decreased MUC5AC, TFF1, and TFF2 expression and increased MUC6 and MUC5B expression. Endocrine cells expressing chromagranin A and gastrin shifted toward the surface epithelium of the antral glands in H. pylori-infected patients. H. pylori infection and concomitant inflammation induced increased epithelial proliferation and triggered coordinate deregulation of secretory cell populations in the antrum. In particular, infection led to a coordinated increase in cells expressing MUC6 and MUC5B at the expense of MUC5AC-producing cells.
