Inhibition of Notch1 signaling has been shown to promote astrocyte-derived neurogenesis after stroke.To investigate the regulatory role of Notch1 signaling in this process,in this study,we used a rat model of stroke b...Inhibition of Notch1 signaling has been shown to promote astrocyte-derived neurogenesis after stroke.To investigate the regulatory role of Notch1 signaling in this process,in this study,we used a rat model of stroke based on middle cerebral artery occlusion and assessed the behavior of reactive astrocytes post-stroke.We used theγ-secretase inhibitor N-[N-(3,5-diuorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester(DAPT)to block Notch1 signaling at 1,4,and 7 days after injury.Our results showed that only administration of DAPT at 4 days after stroke promoted astrocyte-derived neurogenesis,as manifested by recovery of white matter fiber bundle integrity on magnetic resonance imaging,which is consistent with recovery of neurologic function.These findings suggest that inhibition of Notch1 signaling at the subacute stage post-stroke mediates neural repair by promoting astrocyte-derived neurogenesis.展开更多
Objective:To study the impact of the Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway on macrophage polarization and its role in modulating the inflammatory response in rats with adjuvant arthritis(AA).Methods:The rats w...Objective:To study the impact of the Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway on macrophage polarization and its role in modulating the inflammatory response in rats with adjuvant arthritis(AA).Methods:The rats were randomly divided into three groups(6 rats):the healthy group(NC),the model group(MC),and the Notch1 inhibitor group(FLI).Medication was administered after 12 days of inducing inflammation.After 30 days,the arthritis index(AI)and degree of swelling in the right hind foot joint(E)were measured in each group.The expression levels of CD80^(+)and CD163^(+)cells in peripheral blood macrophages of rats were analyzed by flow cytometry.The standards of IL-4,IL-10,IL-1β,and TNF-α in rat serum were gauged by Enzyme-linked immunosorbent assay.The expression of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in rat synovial tissue was detected using Western blot.Results:The degree of swelling(E)and arthritis index(AI)in the MC group rats with AA were significantly higher than those in the NC group(P<0.01).CD80^(+)cell expression was significantly higher compared to the control group(P<0.01),while CD163^(+)cell expression was significantly lower than the control group(P<0.01).IL-1βand TNF-α expression levels were significantly elevated(P<0.01),whereas IL-4 and IL-10 expression levels were significantly decreased(P<0.01).Notch1,RBP-Jκ,Jagged1,and Hes1 protein expression levels were significantly increased(P<0.01).In comparison to the MC group,the rats in the Notch1 inhibitor group exhibited a significant reduction in toe swelling and arthritis index(P<0.01).CD80^(+)cell expression was significantly decreased(P<0.01),while CD163+cell expression was significantly increased(P<0.01).IL-1β and TNF-α expression levels were significantly decreased(P<0.05),whereas IL-4 and IL-10 levels were significantly increased(P<0.01).Notch1,Jagged1,Hes1,and RBP-Jκ protein expression levels were significantly decreased(P<0.05).Correlation analysis revealed a positive association between CD80^(+)and Notch1,Jagged1,Hes1,and RBP-Jκ(P<0.01),while CD163^(+)showed a negative correlation with the expression of these proteins(P<0.01).Conclusion:The Notch1/Jagged1/RBP-Jκ/Hes1 signaling axis regulates macrophage polarization to M2 type and reduces inflammation in AA rats.展开更多
目的探讨果蝇双翅边缘缺刻同源基因(Notch)信号通路在慢性阻塞性肺疾病(COPD)辅助性T细胞1(Helper T cells 1,Th1)和辅助性T细胞2(Helper T cells 2,Th2)失衡中的作用及芪蛭皱肺颗粒的干预机制。方法70只Wistar大鼠随机挑选10只作为空...目的探讨果蝇双翅边缘缺刻同源基因(Notch)信号通路在慢性阻塞性肺疾病(COPD)辅助性T细胞1(Helper T cells 1,Th1)和辅助性T细胞2(Helper T cells 2,Th2)失衡中的作用及芪蛭皱肺颗粒的干预机制。方法70只Wistar大鼠随机挑选10只作为空白对照组,其余大鼠均采用香烟烟雾(CS)联合气管滴注脂多糖(Lipopolysaccharide,LPS)法建立COPD模型,空白对照组及造模组各随机挑选3只大鼠验证造模是否成功。造模结束进行灌胃给药干预,造模组大鼠随机分为模型对照组、阳性对照组(67.5μg·kg^(-1))及芪蛭皱肺颗粒高中低剂量组(3.24、1.62、0.81 g·kg^(-1)),分别给予生理盐水、醋酸地塞米松混悬液、芪蛭皱肺高、中、低剂量混悬液进行灌胃干预,空白对照组同模型对照组,灌胃等体积生理盐水。经28天造模及28天治疗后,采用动物肺功能测试系统检测吸气峰流速(Peak Inspiratory Flow,PIF)和呼气峰流速(Peak Expiratory Flow,PEF),处死大鼠提取肺脏、脾脏、血清及支气管肺泡灌洗液(BALF),苏木素-伊红(HE)染色评价肺组织病理变化,酶联免疫吸附实验法(ELISA)测定血清及BALF中肿瘤坏死因子-α(TNF-α)含量,流式细胞仪检测脾脏Th1/Th2细胞水平,免疫组织化学法(Immunohistochemistry,IHC)及蛋白免疫印迹法(Western blot)检测肺组织Notch1、Hes家族发状分裂相关增强子1(Hes1)、Hey家族发状分裂相关增强子1(Hey1)蛋白水平,实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)检测肺组织Notch1、Hes1、Hey1基因表达水平。结果与空白对照组比较,模型对照组大鼠肺功能显著降低(P<0.05),肺组织出现炎性细胞浸润、支气管结构破坏等病变,血清及BALF中TNF-α含量显著升高(P<0.05),脾Th1细胞百分比显著降低(P<0.05),Th2细胞百分比显著升高(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著升高(P<0.05),差异均具有统计学意义;与模型对照组比较,各给药组大鼠肺功能显著升高(P<0.05),肺组织病理损伤均有所减轻,血清及BALF中TNF-α含量显著降低(P<0.05),脾Th1细胞百分比显著升高(P<0.05),Th2细胞百分比显著降低(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著降低(P<0.05),差异均具有统计学意义。结论芪蛭皱肺颗粒通过抑制Notch信号通路调节Th1/Th2平衡,从而改善COPD大鼠肺功能及病理损伤,影响其免疫功能。展开更多
基金supported by the National Natural Science Foundation of China,Nos.81801660(to XZH)and 81771788(to YMY)。
文摘Inhibition of Notch1 signaling has been shown to promote astrocyte-derived neurogenesis after stroke.To investigate the regulatory role of Notch1 signaling in this process,in this study,we used a rat model of stroke based on middle cerebral artery occlusion and assessed the behavior of reactive astrocytes post-stroke.We used theγ-secretase inhibitor N-[N-(3,5-diuorophenacetyl)-1-alanyl]-S-phenylglycine t-butylester(DAPT)to block Notch1 signaling at 1,4,and 7 days after injury.Our results showed that only administration of DAPT at 4 days after stroke promoted astrocyte-derived neurogenesis,as manifested by recovery of white matter fiber bundle integrity on magnetic resonance imaging,which is consistent with recovery of neurologic function.These findings suggest that inhibition of Notch1 signaling at the subacute stage post-stroke mediates neural repair by promoting astrocyte-derived neurogenesis.
基金National Natural Science Foundation General Project(No.81973655,82274501)The Seventh Batch of National Elderly Traditional Chinese Medicine Experts'Academic Experience Inheritance Project of the National Administration of Traditional Chinese Medicine(No.Guo Zhong Yao Ren Jiao Han[2022]No.76)+3 种基金Anhui Provincial Excellent Youth Talent Support Program for Universities(No.Anhui Education Secretary[2022]No.11)Key Laboratory of Xin'an Medical Education Department(No.2020xayx04)Excellent Youth Talent Support Program for Universities(No.Anhui Education Secretary[2022]No.11)Provincial Education and Teaching Reform Research Project of Anhui Province Higher Education Quality Engineering Project(No.2022jyxm883)。
文摘Objective:To study the impact of the Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway on macrophage polarization and its role in modulating the inflammatory response in rats with adjuvant arthritis(AA).Methods:The rats were randomly divided into three groups(6 rats):the healthy group(NC),the model group(MC),and the Notch1 inhibitor group(FLI).Medication was administered after 12 days of inducing inflammation.After 30 days,the arthritis index(AI)and degree of swelling in the right hind foot joint(E)were measured in each group.The expression levels of CD80^(+)and CD163^(+)cells in peripheral blood macrophages of rats were analyzed by flow cytometry.The standards of IL-4,IL-10,IL-1β,and TNF-α in rat serum were gauged by Enzyme-linked immunosorbent assay.The expression of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in rat synovial tissue was detected using Western blot.Results:The degree of swelling(E)and arthritis index(AI)in the MC group rats with AA were significantly higher than those in the NC group(P<0.01).CD80^(+)cell expression was significantly higher compared to the control group(P<0.01),while CD163^(+)cell expression was significantly lower than the control group(P<0.01).IL-1βand TNF-α expression levels were significantly elevated(P<0.01),whereas IL-4 and IL-10 expression levels were significantly decreased(P<0.01).Notch1,RBP-Jκ,Jagged1,and Hes1 protein expression levels were significantly increased(P<0.01).In comparison to the MC group,the rats in the Notch1 inhibitor group exhibited a significant reduction in toe swelling and arthritis index(P<0.01).CD80^(+)cell expression was significantly decreased(P<0.01),while CD163+cell expression was significantly increased(P<0.01).IL-1β and TNF-α expression levels were significantly decreased(P<0.05),whereas IL-4 and IL-10 levels were significantly increased(P<0.01).Notch1,Jagged1,Hes1,and RBP-Jκ protein expression levels were significantly decreased(P<0.05).Correlation analysis revealed a positive association between CD80^(+)and Notch1,Jagged1,Hes1,and RBP-Jκ(P<0.01),while CD163^(+)showed a negative correlation with the expression of these proteins(P<0.01).Conclusion:The Notch1/Jagged1/RBP-Jκ/Hes1 signaling axis regulates macrophage polarization to M2 type and reduces inflammation in AA rats.
