Notch pathway activation maintains neural stem cells in a proliferating state and increases nerve repair capacity. To date, studies have rarely focused on changes or damage to signal transduc- tion pathways during cer...Notch pathway activation maintains neural stem cells in a proliferating state and increases nerve repair capacity. To date, studies have rarely focused on changes or damage to signal transduc- tion pathways during cerebral hemorrhage. Here, we examined the effect of acupuncture in a rat model of cerebral hemorrhage. We examined four groups： in the control group, rats received no treatment. In the model group, cerebral hemorrhage models were established by infusing non-hep-arinized blood into the Brain. In the acupuncture group, modeled rats had Baihui （DU20） and Qubin （GBT） acupoints treated once a day for 30 minutes. In the DAPT group, modeled rats had 0.15 μg/mL DAPT solution （10 mL） infused into the brain. Immunohistochemistry and western blot results showed that acupuncture effectively inhibits Notch 1 and Hesl protein expression in rat basal ganglia. These inhibitory effects were identical to DAPT, a Notch signaling pathway inhibitor. Our results suggest that acupuncture has a neuroprotective effect on cerebral hemorrhage by in- hibiting Notch-Hes signaling pathway transduction in rat basal ganglia after cerebral hemorrhage.展开更多
AIM:To determine the role of Notch1 and Hes1 in regulating the activation of hepatic stellate cells(HSCs) and whether Hes1 is regulated by transforming growth factor(TGF)/bone morphogenetic protein(BMP) signaling.METH...AIM:To determine the role of Notch1 and Hes1 in regulating the activation of hepatic stellate cells(HSCs) and whether Hes1 is regulated by transforming growth factor(TGF)/bone morphogenetic protein(BMP) signaling.METHODS:Immunofluorescence staining was used to detect the expression of desmin,glial fibrillary acidic protein and the myofibroblastic marker α-smooth muscle actin(α-SMA) after freshly isolated,normal rat HSCs had been activated in culture for different numbers of days(0,1,3,7 and 10 d).The expression of α-SMA,collagen1α2(COL1α2),Notch receptors(Notch1-4),and the Notch target genes Hes1 and Hey1 were analyzed by reverse transcriptase-polymerase chain reaction.Luciferase reporter assays and Western blot were used to study the regulation of α-SMA,COL1α1,COL1α2 and Hes1 by NICD1,Hes1,CA-ALK3,and CA-ALK5 in HSC-T6 cells.Moreover,the effects of inhibiting Hes1 function in HSC-T6 cells using a Hes1 decoy were also investigated.RESULTS:The expression of Notch1 and Hes1 m RNAs was significantly down-regulated during the culture of freshly isolated HSCs.In HSC-T6 cells,Notch1 inhibited the promoter activities of α-SMA,COL1α1 and COL1α2.On the other hand,Hes1 enhanced the promoter activities of α-SMA and COL1α2,and this effect could be blocked by inhibiting Hes1 function with a Hes1 decoy.Furthermore,co-transfection of pc DNA3-CAALK3(BMP signaling activin receptor-like kinase 3) and pc DNA3.1-NICD1 further increased the expression of Hes1 compared with transfection of either vector alone in HSC-T6 cells,while pc DNA3-CA-ALK5(TGF-β signaling activin receptor-like kinase 5) reduced the effect of NICD1 on Hes1 expression.CONCLUSION:Selective interruption of Hes1 or maintenance of Hes1 at a reasonable level decreases the promoter activities of α-SMA and COL1α2,and these conditions may provide an anti-fibrotic strategy against hepatic fibrosis.展开更多
AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) express...AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) expression in hepatoma samples was confirmed by q RT-PCR. In addition, MTT assays, flow cytometry and TUNEL analysis showed that DHM possessed strong anti-tumor properties, evidenced not only by reduced cell proliferation but also by enhanced apoptosis in QGY7701 and Hep G2 hepatocellular carcinoma(HCC) cells. The expressions of Notch1, Hes1, Bcl-2 and Bax were determined by Western blot.RESULTS Among the tested samples(n = 64), the expression levels of Notch1(75% of patients) and Hes1(79.7% of patients) m RNA in tumor tissues were higher than in the normal liver tissues. There was a negative correlation between the expression of Notch1 and the degree of differentiation and positively correlated with the Alpha Fetal Protein concentration. The viability of HCC cells treated with DHM was significantly inhibited in a dose and time-dependent manner. Apoptosis was induced in Hep G2 and QGY7701 cell lines following 24 h of DHM treatment. After treatment with DHM, the protein expression of Notch1 was downregulated, the apoptosis-related protein Bax was upregulated and Bcl2 was downregulated. Notch1 si RNA further enhanced the anti-tumor properties of DHM. CONCLUSION Notch1 is involved in the development of HCC and DHM inhibits cell proliferation and promotes apoptosis by down-regulating the expression of Notch1.展开更多
As an inhibitor of the Notch signaling pathway, N-[N-(3,5-difluorohenacetyl)-l-alanyl]-S-phenylglycine tert-butyl ester(DAPT) may protect brain tissue from serious ischemic injury. This study aimed to explore neuropro...As an inhibitor of the Notch signaling pathway, N-[N-(3,5-difluorohenacetyl)-l-alanyl]-S-phenylglycine tert-butyl ester(DAPT) may protect brain tissue from serious ischemic injury. This study aimed to explore neuroprotection by DAPT after cerebral ischemia/reperfusion(I/R) injury. DAPT was intraperitoneally injected 3 hours before the establishment of a focal cerebral I/R model in the right middle cerebral artery of obstructed mice. Longa scores were used to assess neurological changes of mice. Nissl staining and TdT-mediated dUTP-biotin nick-end labeling staining were used to examine neuronal damage and cell apoptosis in the right prefrontal cortex, while immunofluorescence staining was used to detect glial fibrillary acidic protein-and Notch1-positive cells. Protein expression levels of Hes1 and Hes5 were detected by western blot assay in the right prefrontal cortex. Our results demonstrated that DAPT significantly improved neurobehavioral scores and relieved neuronal morphological damage. DAPT decreased the number of glial fibrillary acidic protein-and Notch1-positive cells in the right prefrontal cortex, while also reducing the number of apoptotic cells and decreasing interleukin-6 and tumor necrosis factor-α contents, and simultaneously downregulating Hes1 and Hes5 protein expression. These findings verify that DAPT alleviates pathological lesions and strengthens the anti-inflammatory response after cerebral I/R injury. Thus, DAPT might be developed as an effective drug for the prevention of cerebral I/R injury.展开更多
基金the Science and Technology Foundation of Henan Province(No.0424410085)the Natural Science Foundation of Henan Province (No.0611043000)
文摘本研究探讨Notch信号通路在人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)体外增殖及向神经细胞分化过程中的作用。采集健康自愿者骨髓,体外培养获得hMSCs,取第3代hMSCs,在诱导剂(β-ME,DMSO,BHA)作用下向神经细胞分化。诱导后用免疫细胞化学鉴定神经元特异性烯醇化酶(neuron-specific enolase,NSE)和尼氏体的表达以确定诱导效果;用流式细胞术检测细胞生长周期时相的变化。在诱导前后,用免疫荧光和RT-PCR方法检测Notch通路中Notch1受体蛋白、配体Jagged1(JAG1)、调节蛋白活化相关物早老素1(presenilin1,PS1)、靶基因hairy and enhancer of split1 (HES1)信号分子表达的变化。结果显示:诱导前,处于G0/G1期的hMSCs占58.5%,S+G2/M期的细胞占41.5%;诱导后,G0/G1期细胞比例升高,而S+G2/M期细胞比例下降,NSE阳性细胞率达(77±0.35)%,细胞质中可见深蓝色的块状或颗粒状尼氏体。免疫荧光显示,诱导前后hMSCs内Notch1和JAG1均呈阳性表达,但RT-PCR检测发现诱导后Notch1、JAG1、PS1和HES1mRNA表达量较诱导前明显降低(均P<0.05)。结果表明,诱导hMSCs向神经细胞分化能抑制Notch信号分子表达,低水平的Notch信号激活可能有利于神经细胞的分化。
基金supported by the National Natural Science Foundation of China,No.81273824,30772840Ministry of Education Doctoral Fund in China,No.20102327110003+1 种基金the Natural Science Foundation of Heilongjiang Province in China,No.ZD201204Special funds for Technological Innovation Research of Harbin,China,No.2012RFXXS062
文摘Notch pathway activation maintains neural stem cells in a proliferating state and increases nerve repair capacity. To date, studies have rarely focused on changes or damage to signal transduc- tion pathways during cerebral hemorrhage. Here, we examined the effect of acupuncture in a rat model of cerebral hemorrhage. We examined four groups： in the control group, rats received no treatment. In the model group, cerebral hemorrhage models were established by infusing non-hep-arinized blood into the Brain. In the acupuncture group, modeled rats had Baihui （DU20） and Qubin （GBT） acupoints treated once a day for 30 minutes. In the DAPT group, modeled rats had 0.15 μg/mL DAPT solution （10 mL） infused into the brain. Immunohistochemistry and western blot results showed that acupuncture effectively inhibits Notch 1 and Hesl protein expression in rat basal ganglia. These inhibitory effects were identical to DAPT, a Notch signaling pathway inhibitor. Our results suggest that acupuncture has a neuroprotective effect on cerebral hemorrhage by in- hibiting Notch-Hes signaling pathway transduction in rat basal ganglia after cerebral hemorrhage.
基金Supported by National Natural Science Foundation of China,No.81170412,No.81070348 and No.81200307Health Department of Hubei Province of China,No.JX6C-26
文摘AIM:To determine the role of Notch1 and Hes1 in regulating the activation of hepatic stellate cells(HSCs) and whether Hes1 is regulated by transforming growth factor(TGF)/bone morphogenetic protein(BMP) signaling.METHODS:Immunofluorescence staining was used to detect the expression of desmin,glial fibrillary acidic protein and the myofibroblastic marker α-smooth muscle actin(α-SMA) after freshly isolated,normal rat HSCs had been activated in culture for different numbers of days(0,1,3,7 and 10 d).The expression of α-SMA,collagen1α2(COL1α2),Notch receptors(Notch1-4),and the Notch target genes Hes1 and Hey1 were analyzed by reverse transcriptase-polymerase chain reaction.Luciferase reporter assays and Western blot were used to study the regulation of α-SMA,COL1α1,COL1α2 and Hes1 by NICD1,Hes1,CA-ALK3,and CA-ALK5 in HSC-T6 cells.Moreover,the effects of inhibiting Hes1 function in HSC-T6 cells using a Hes1 decoy were also investigated.RESULTS:The expression of Notch1 and Hes1 m RNAs was significantly down-regulated during the culture of freshly isolated HSCs.In HSC-T6 cells,Notch1 inhibited the promoter activities of α-SMA,COL1α1 and COL1α2.On the other hand,Hes1 enhanced the promoter activities of α-SMA and COL1α2,and this effect could be blocked by inhibiting Hes1 function with a Hes1 decoy.Furthermore,co-transfection of pc DNA3-CAALK3(BMP signaling activin receptor-like kinase 3) and pc DNA3.1-NICD1 further increased the expression of Hes1 compared with transfection of either vector alone in HSC-T6 cells,while pc DNA3-CA-ALK5(TGF-β signaling activin receptor-like kinase 5) reduced the effect of NICD1 on Hes1 expression.CONCLUSION:Selective interruption of Hes1 or maintenance of Hes1 at a reasonable level decreases the promoter activities of α-SMA and COL1α2,and these conditions may provide an anti-fibrotic strategy against hepatic fibrosis.
基金Supported by The National Natural Science Foundation of China,No.81041099Natural Science Foundation of Guangdong Province,China,No.S2011010003750
文摘AIM To investigate whether Dihydromyricetin(DHM) inhibits cell proliferation and promotes apoptosis by downregulating Notch1 expression.METHODS The correlation between Notch1 and Hes1(a Notch1 target molecule) expression in hepatoma samples was confirmed by q RT-PCR. In addition, MTT assays, flow cytometry and TUNEL analysis showed that DHM possessed strong anti-tumor properties, evidenced not only by reduced cell proliferation but also by enhanced apoptosis in QGY7701 and Hep G2 hepatocellular carcinoma(HCC) cells. The expressions of Notch1, Hes1, Bcl-2 and Bax were determined by Western blot.RESULTS Among the tested samples(n = 64), the expression levels of Notch1(75% of patients) and Hes1(79.7% of patients) m RNA in tumor tissues were higher than in the normal liver tissues. There was a negative correlation between the expression of Notch1 and the degree of differentiation and positively correlated with the Alpha Fetal Protein concentration. The viability of HCC cells treated with DHM was significantly inhibited in a dose and time-dependent manner. Apoptosis was induced in Hep G2 and QGY7701 cell lines following 24 h of DHM treatment. After treatment with DHM, the protein expression of Notch1 was downregulated, the apoptosis-related protein Bax was upregulated and Bcl2 was downregulated. Notch1 si RNA further enhanced the anti-tumor properties of DHM. CONCLUSION Notch1 is involved in the development of HCC and DHM inhibits cell proliferation and promotes apoptosis by down-regulating the expression of Notch1.
基金supported by the National Natural Science Foundation of China,No.81660243(to JDZ)a grant from the Social Development Science and Technology Plan Project of Science and Technology Department of Guizhou Province of China,No.SY [2015] 3041(to JDZ)a grant from the Science and Technology Department of Guizhou Province of China,No.LG [2012] 028(to JDZ)
文摘As an inhibitor of the Notch signaling pathway, N-[N-(3,5-difluorohenacetyl)-l-alanyl]-S-phenylglycine tert-butyl ester(DAPT) may protect brain tissue from serious ischemic injury. This study aimed to explore neuroprotection by DAPT after cerebral ischemia/reperfusion(I/R) injury. DAPT was intraperitoneally injected 3 hours before the establishment of a focal cerebral I/R model in the right middle cerebral artery of obstructed mice. Longa scores were used to assess neurological changes of mice. Nissl staining and TdT-mediated dUTP-biotin nick-end labeling staining were used to examine neuronal damage and cell apoptosis in the right prefrontal cortex, while immunofluorescence staining was used to detect glial fibrillary acidic protein-and Notch1-positive cells. Protein expression levels of Hes1 and Hes5 were detected by western blot assay in the right prefrontal cortex. Our results demonstrated that DAPT significantly improved neurobehavioral scores and relieved neuronal morphological damage. DAPT decreased the number of glial fibrillary acidic protein-and Notch1-positive cells in the right prefrontal cortex, while also reducing the number of apoptotic cells and decreasing interleukin-6 and tumor necrosis factor-α contents, and simultaneously downregulating Hes1 and Hes5 protein expression. These findings verify that DAPT alleviates pathological lesions and strengthens the anti-inflammatory response after cerebral I/R injury. Thus, DAPT might be developed as an effective drug for the prevention of cerebral I/R injury.
