Stem cell-based therapies have been proposed as a potential treatment for neural regeneration following closed head injury.We previously reported that induced neural stem cells exert beneficial effects on neural regen...Stem cell-based therapies have been proposed as a potential treatment for neural regeneration following closed head injury.We previously reported that induced neural stem cells exert beneficial effects on neural regeneration via cell replacement.However,the neural regeneration efficiency of induced neural stem cells remains limited.In this study,we explored differentially expressed genes and long non-coding RNAs to clarify the mechanism underlying the neurogenesis of induced neural stem cells.We found that H19 was the most downregulated neurogenesis-associated lnc RNA in induced neural stem cells compared with induced pluripotent stem cells.Additionally,we demonstrated that H19 levels in induced neural stem cells were markedly lower than those in induced pluripotent stem cells and were substantially higher than those in induced neural stem cell-derived neurons.We predicted the target genes of H19 and discovered that H19 directly interacts with mi R-325-3p,which directly interacts with Ctbp2 in induced pluripotent stem cells and induced neural stem cells.Silencing H19 or Ctbp2 impaired induced neural stem cell proliferation,and mi R-325-3p suppression restored the effect of H19 inhibition but not the effect of Ctbp2 inhibition.Furthermore,H19 silencing substantially promoted the neural differentiation of induced neural stem cells and did not induce apoptosis of induced neural stem cells.Notably,silencing H19 in induced neural stem cell grafts markedly accelerated the neurological recovery of closed head injury mice.Our results reveal that H19 regulates the neurogenesis of induced neural stem cells.H19 inhibition may promote the neural differentiation of induced neural stem cells,which is closely associated with neurological recovery following closed head injury.展开更多
基金supported by the National Natural Science Foundation of China,Nos.82271397(to MG),82001293(to MG),82171355(to RX),81971295(to RX)and 81671189(to RX)。
文摘Stem cell-based therapies have been proposed as a potential treatment for neural regeneration following closed head injury.We previously reported that induced neural stem cells exert beneficial effects on neural regeneration via cell replacement.However,the neural regeneration efficiency of induced neural stem cells remains limited.In this study,we explored differentially expressed genes and long non-coding RNAs to clarify the mechanism underlying the neurogenesis of induced neural stem cells.We found that H19 was the most downregulated neurogenesis-associated lnc RNA in induced neural stem cells compared with induced pluripotent stem cells.Additionally,we demonstrated that H19 levels in induced neural stem cells were markedly lower than those in induced pluripotent stem cells and were substantially higher than those in induced neural stem cell-derived neurons.We predicted the target genes of H19 and discovered that H19 directly interacts with mi R-325-3p,which directly interacts with Ctbp2 in induced pluripotent stem cells and induced neural stem cells.Silencing H19 or Ctbp2 impaired induced neural stem cell proliferation,and mi R-325-3p suppression restored the effect of H19 inhibition but not the effect of Ctbp2 inhibition.Furthermore,H19 silencing substantially promoted the neural differentiation of induced neural stem cells and did not induce apoptosis of induced neural stem cells.Notably,silencing H19 in induced neural stem cell grafts markedly accelerated the neurological recovery of closed head injury mice.Our results reveal that H19 regulates the neurogenesis of induced neural stem cells.H19 inhibition may promote the neural differentiation of induced neural stem cells,which is closely associated with neurological recovery following closed head injury.
文摘目的:分析lncRNA-H19(H19)在高血压脑出血(HICH)患者体内的表达特征,并探讨H19影响人脑微血管内皮细胞(HBMECs)自噬与氧糖剥夺(OGD)损伤的机制。方法:收集健康者、高血压患者和HICH患者血液样本,采用lncRNA array芯片检测lncRNA表达谱,RT-qPCR检测H19表达水平并分析其与脑出血量及NIHSS评分的关系;预测H19的靶蛋白,RT-qPCR检测其mRNA水平并分析其与H19的相关关系;采用OGD处理敲降或过表达H19及其互作蛋白的HBMECs,而后采用免疫荧光检测细胞自噬情况,western blotting检测自噬相关蛋白(LC3-Ⅰ和LC3-Ⅱ)以及缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)。结果:H19在高血压患者(t=0.021,P<0.05)和HICH患者(t=0.018,P<0.05)中显著高于健康者;并随HICH分级升高而增加,与脑出血量及NIHSS评分呈正相关关系(R2=0.087,R2=0.113,P<0.05);随OGD处理时间增加,HBMECs中H19水平增加(t=0.023,P<0.05);IGF1R m RNA在HICH患者体内低于健康者(t=0.015,P<0.05),且随HICH加剧而降低(t=0.043,P<0.05);HICH患者体内H19与IGF1R呈负相关关系(R2=0.187,P<0.05);OGD处理下调HBMECs中IGF1R蛋白,敲降H19可使OGD-HBMECs中IGF1R增加;在OGD-HBMECs中LC3自噬体、LC3-Ⅱ/LC3-Ⅰ、HIF-1α和VEGF均因敲降H19或过表达IGF1R而减少,但同时过表达IGF1R及H19后则均增加。结论:H19与HICH病情相关,下调H19可增加IGF1R,进而缓解OGD诱导的HBMECs自噬和损伤。
