Objective Our aim was to explore whether heat stress protein(HSP) 9 preferentially expresses under heat stress and affects the expression of other heat stress proteins as well as to explore the effect of HSPB9 overexp...Objective Our aim was to explore whether heat stress protein(HSP) 9 preferentially expresses under heat stress and affects the expression of other heat stress proteins as well as to explore the effect of HSPB9 overexpression and knockdown on apoptosis in DF-1. Methods We used gene cloning to construct an overexpression vector of the target gene, and synthesized the target gene interference fragment to transfect the chicken fibroblast cell line. Gene and protein expression, as well as apoptosis, were detected by RT-qPCR, Western blot, and flow cytometry. Results Chicken DF-1 cells showed an early state of apoptosis in the early stages of HSPB9 overexpression. In the later stages, as HSPB9 expression increased, the cells showed inhibition of apoptosis. When the cells were under heat stress, HSPB9 expression was much higher and earlier than the expression of HSPB1 and HSPA2. In addition, high expression of HSPB9 had a negative effect on HSPB1 and HSPA2 expression. This negative feedback decreased the percentage of early stages of apoptotic cells and promoted cell survival. Conclusion HSPB9 expression, although rapid, is detrimental to cell survival early during its overexpression. In heat stress, HSPB9 overexpression, while inhibiting the expression of HSPA2 and HSPB1, is beneficial to cell survival.展开更多
OBJECTIVE To investigate the role of adventitial vasa vasorum in artery remodeling during the process of pulmonary artery hypertension(PAH),we checked the small heat shock protein 27/25(HSPB1)whether involved in patho...OBJECTIVE To investigate the role of adventitial vasa vasorum in artery remodeling during the process of pulmonary artery hypertension(PAH),we checked the small heat shock protein 27/25(HSPB1)whether involved in pathological basis of vascular remodeling.METHODS We explored the potential role of HSPB1 interacts with ectopic F1Fo-ATPase in the pulmonary vascular remodeling,investigate its effects on the endothelium cell dynamic,and further reveal its possible molecular mechanisms using hypoxic pulmonary hypertension rat model,transgenic mice and pulmonary adventitial vasa vasorum endothelial cell culture in vitro.RESULTS Our studies have shown that HSPB1 improves adventitial vasa vasorum angiogenesis and remodeling.We found that hypoxia induces-HSPB1 upregulation and HSPB1 interact with ectopic F1Fo-ATPase modulate adventitial vasa vasorum endothelial cell proliferation,migration and tube formation.And the inhibition of HSPB1can reverse the vascular inflammation and fibrosis amazingly.CONCLUSION Adventitial vasa vasorum plays an important role in vascular remodeling,and small heat shock protein 27/25 was involved in a variety of diseases during the development of PAH,which could an efficient therapeutic targets and prevention strategy for PAH clinical.展开更多
目的探讨轴突型腓骨肌萎缩症2L型(axonal Charcot-Marie-Tooth disease type 2L,CMT2L)致病基因小分子热休克蛋白HSPB8(smallheatshockproteinHSPB8,HSPB8)的K141N突变导致细胞内聚集物形成的可能机理。方法建立pEGFPN1-HSPB8、pEGFPN1-...目的探讨轴突型腓骨肌萎缩症2L型(axonal Charcot-Marie-Tooth disease type 2L,CMT2L)致病基因小分子热休克蛋白HSPB8(smallheatshockproteinHSPB8,HSPB8)的K141N突变导致细胞内聚集物形成的可能机理。方法建立pEGFPN1-HSPB8、pEGFPN1-K141NHSPB8瞬时表达细胞模型,并进行EGFP-K141NHSPB8与小分子热休克蛋白HSPB1(smallheatshockproteinHSPB1,HSPB1)、神经丝轻链(neurofilamentlightchain,NEFL)的免疫荧光共定位分析,观察EGFP-K141NHSPB8在不同内源性表达细胞系的聚集物形成情况,采用t检验和单因素方差分析的统计学方法分析聚集物形成的可能机理。结果EGFP-K141NHSPB8形成以核周分布为主的聚集物,EGFP-K141NHSPB8与HSPB1、NEFL均存在免疫荧光共定位。EGFP-K141NHSPB8在不同内源性表达细胞系的聚集物形成百分率的差异有统计学意义。结论突变型HSPB8(K141N)形成以核周分布为主的胞内聚集物,聚集物中K141NHSPB8与HSPB1、NEFL均存在共定位。聚集物形成的可能机理包括K141NHSPB8多肽链构象发生改变后不能维持稳态而出现自身异常聚集;与家族内其他成员特别是HSPB1结合成异常的异源多聚体,在胞内形成不可溶性大分子后产生聚集。展开更多
基金supported by grants from the National Science and Technology Support Program of China [2014BAD08B08]National Natural Science Foundation of China [30972093]
文摘Objective Our aim was to explore whether heat stress protein(HSP) 9 preferentially expresses under heat stress and affects the expression of other heat stress proteins as well as to explore the effect of HSPB9 overexpression and knockdown on apoptosis in DF-1. Methods We used gene cloning to construct an overexpression vector of the target gene, and synthesized the target gene interference fragment to transfect the chicken fibroblast cell line. Gene and protein expression, as well as apoptosis, were detected by RT-qPCR, Western blot, and flow cytometry. Results Chicken DF-1 cells showed an early state of apoptosis in the early stages of HSPB9 overexpression. In the later stages, as HSPB9 expression increased, the cells showed inhibition of apoptosis. When the cells were under heat stress, HSPB9 expression was much higher and earlier than the expression of HSPB1 and HSPA2. In addition, high expression of HSPB9 had a negative effect on HSPB1 and HSPA2 expression. This negative feedback decreased the percentage of early stages of apoptotic cells and promoted cell survival. Conclusion HSPB9 expression, although rapid, is detrimental to cell survival early during its overexpression. In heat stress, HSPB9 overexpression, while inhibiting the expression of HSPA2 and HSPB1, is beneficial to cell survival.
文摘OBJECTIVE To investigate the role of adventitial vasa vasorum in artery remodeling during the process of pulmonary artery hypertension(PAH),we checked the small heat shock protein 27/25(HSPB1)whether involved in pathological basis of vascular remodeling.METHODS We explored the potential role of HSPB1 interacts with ectopic F1Fo-ATPase in the pulmonary vascular remodeling,investigate its effects on the endothelium cell dynamic,and further reveal its possible molecular mechanisms using hypoxic pulmonary hypertension rat model,transgenic mice and pulmonary adventitial vasa vasorum endothelial cell culture in vitro.RESULTS Our studies have shown that HSPB1 improves adventitial vasa vasorum angiogenesis and remodeling.We found that hypoxia induces-HSPB1 upregulation and HSPB1 interact with ectopic F1Fo-ATPase modulate adventitial vasa vasorum endothelial cell proliferation,migration and tube formation.And the inhibition of HSPB1can reverse the vascular inflammation and fibrosis amazingly.CONCLUSION Adventitial vasa vasorum plays an important role in vascular remodeling,and small heat shock protein 27/25 was involved in a variety of diseases during the development of PAH,which could an efficient therapeutic targets and prevention strategy for PAH clinical.
文摘目的探讨轴突型腓骨肌萎缩症2L型(axonal Charcot-Marie-Tooth disease type 2L,CMT2L)致病基因小分子热休克蛋白HSPB8(smallheatshockproteinHSPB8,HSPB8)的K141N突变导致细胞内聚集物形成的可能机理。方法建立pEGFPN1-HSPB8、pEGFPN1-K141NHSPB8瞬时表达细胞模型,并进行EGFP-K141NHSPB8与小分子热休克蛋白HSPB1(smallheatshockproteinHSPB1,HSPB1)、神经丝轻链(neurofilamentlightchain,NEFL)的免疫荧光共定位分析,观察EGFP-K141NHSPB8在不同内源性表达细胞系的聚集物形成情况,采用t检验和单因素方差分析的统计学方法分析聚集物形成的可能机理。结果EGFP-K141NHSPB8形成以核周分布为主的聚集物,EGFP-K141NHSPB8与HSPB1、NEFL均存在免疫荧光共定位。EGFP-K141NHSPB8在不同内源性表达细胞系的聚集物形成百分率的差异有统计学意义。结论突变型HSPB8(K141N)形成以核周分布为主的胞内聚集物,聚集物中K141NHSPB8与HSPB1、NEFL均存在共定位。聚集物形成的可能机理包括K141NHSPB8多肽链构象发生改变后不能维持稳态而出现自身异常聚集;与家族内其他成员特别是HSPB1结合成异常的异源多聚体,在胞内形成不可溶性大分子后产生聚集。
