目的探讨银杏叶提取物EGB761对糖尿病肾病(DKD)小鼠肾小管损伤及内质网应激的影响。方法将50只C57BL/6N小鼠随机分为正常组、DKD模型组、DKD+EGB761组(36 mg·kg-1)以及DKD+4-苯基丁酸(4-PBA)组(1 g·kg-1)。通过高脂饲料联合...目的探讨银杏叶提取物EGB761对糖尿病肾病(DKD)小鼠肾小管损伤及内质网应激的影响。方法将50只C57BL/6N小鼠随机分为正常组、DKD模型组、DKD+EGB761组(36 mg·kg-1)以及DKD+4-苯基丁酸(4-PBA)组(1 g·kg-1)。通过高脂饲料联合腹腔注射链脲佐菌素(STZ)制备糖尿病小鼠模型,糖尿病模型复制成功并继续高脂饲料喂养8周后检测各组尿蛋白,阳性者给药8周,给药期间观察各组小鼠一般情况。给药8周后检测血肌酐(Scr)、尿素氮(BUN)、24 h尿蛋白(24 h Pro)、空腹血糖(FBG)、肾脏肥大指数;HE染色、Masson染色、PAS染色观察各组小鼠肾脏病理改变;透射电镜观察肾小球及肾小管超微结构;ELLSA法检测各组小鼠肾小管损伤标志物尿β2-微球蛋白(β2-MG)、视黄醇结合蛋白4(RBP4)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的表达;以免疫组化、蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Realtime-PCR)检测肾组织内质网应激标志物GRP78、ATF6的表达。结果与正常组比,DKD模型组精神状态较差、反应迟钝、多饮多食多尿症状明显,Scr、BUN、24 h Pro、FBG、肾脏肥大指数均明显升高(P<0.01),肾脏病理损伤加重,尿β2-MG、RBP4、NGAL明显升高(P<0.01),肾组织GRP78、ATF6的表达明显升高(P<0.01)。与DKD模型组相比,DKD+EGB761组精神状态、反应情况及多饮多食多尿症状好转,Scr、BUN、24 h Pro、FBG水平均明显降低(P<0.01),肾脏肥大指数明显降低(P<0.05),肾脏病理损伤明显改善,尿β2-MG、RBP4、NGAL表达降低(P<0.05,P<0.01),肾组织GRP78、ATF6蛋白的表达降低(P<0.05,P<0.01)。结论银杏叶提取物EGB761可以改善DKD小鼠肾功能,减少蛋白尿,减轻肾小管损伤,其作用机制可能与抑制内质网应激相关。展开更多
The ginkgo biloba extract EGb761 improves memory loss and cognitive impairments in patients with senile dementia. It also promotes proliferation of neural stem cells in the subventricular zone in Parkinson＇s disease ...The ginkgo biloba extract EGb761 improves memory loss and cognitive impairments in patients with senile dementia. It also promotes proliferation of neural stem cells in the subventricular zone in Parkinson＇s disease model mice and in the hippocampal zone of young epileptic rats. However, it remains unclear whether EGb761 enhances proliferation of endogenous neural stem cells in the brain of rats with vascular dementia. In this study, a vascular dementia model was established by repeatedly clipping and reperfusing the bilateral common carotid arteries of rats in combination with an intraperitoneal injection of a sodium nitroprusside solution. Seven days after establishing the model, rats were intragastrically given EGb761 at 50 mg/kg per day. Learning and memory abilities were assessed using the Morris water maze and proliferation of endogenous neural stem cells in the subventricular zone and dentate gyrus were labeled by 5-bromo-2-deoxyuridine immunofluorescence in all rats at 15 days, and 1, 2, and 4 months after model establishment. The escape latencies in Morris water maze tests of rats with vascular dementia after EGb761 treatment were significantly shorter than the model group. Immunofluorescence staining showed that the number and proliferation of 5-bromo-2-deoxyuridine-positive cells in the subventricular zone and dentate gyrus of the EGb761-treated group were significantly higher than in the model group. These experimental findings suggest that EGb761 enhances proliferation of neural stem cells in the subventricular zone and dentate gyrus, and significantly improves learning and memory in rats with vascular dementia.展开更多
文摘目的探讨银杏叶提取物EGB761对糖尿病肾病(DKD)小鼠肾小管损伤及内质网应激的影响。方法将50只C57BL/6N小鼠随机分为正常组、DKD模型组、DKD+EGB761组(36 mg·kg-1)以及DKD+4-苯基丁酸(4-PBA)组(1 g·kg-1)。通过高脂饲料联合腹腔注射链脲佐菌素(STZ)制备糖尿病小鼠模型,糖尿病模型复制成功并继续高脂饲料喂养8周后检测各组尿蛋白,阳性者给药8周,给药期间观察各组小鼠一般情况。给药8周后检测血肌酐(Scr)、尿素氮(BUN)、24 h尿蛋白(24 h Pro)、空腹血糖(FBG)、肾脏肥大指数;HE染色、Masson染色、PAS染色观察各组小鼠肾脏病理改变;透射电镜观察肾小球及肾小管超微结构;ELLSA法检测各组小鼠肾小管损伤标志物尿β2-微球蛋白(β2-MG)、视黄醇结合蛋白4(RBP4)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的表达;以免疫组化、蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Realtime-PCR)检测肾组织内质网应激标志物GRP78、ATF6的表达。结果与正常组比,DKD模型组精神状态较差、反应迟钝、多饮多食多尿症状明显,Scr、BUN、24 h Pro、FBG、肾脏肥大指数均明显升高(P<0.01),肾脏病理损伤加重,尿β2-MG、RBP4、NGAL明显升高(P<0.01),肾组织GRP78、ATF6的表达明显升高(P<0.01)。与DKD模型组相比,DKD+EGB761组精神状态、反应情况及多饮多食多尿症状好转,Scr、BUN、24 h Pro、FBG水平均明显降低(P<0.01),肾脏肥大指数明显降低(P<0.05),肾脏病理损伤明显改善,尿β2-MG、RBP4、NGAL表达降低(P<0.05,P<0.01),肾组织GRP78、ATF6蛋白的表达降低(P<0.05,P<0.01)。结论银杏叶提取物EGB761可以改善DKD小鼠肾功能,减少蛋白尿,减轻肾小管损伤,其作用机制可能与抑制内质网应激相关。
基金financially sponsored by the Natural Science Foundation of Shandong Province,No.Y2008C32Scientific Research Funds of Shandong Provincial Education Ministry,No.J01K09
文摘The ginkgo biloba extract EGb761 improves memory loss and cognitive impairments in patients with senile dementia. It also promotes proliferation of neural stem cells in the subventricular zone in Parkinson＇s disease model mice and in the hippocampal zone of young epileptic rats. However, it remains unclear whether EGb761 enhances proliferation of endogenous neural stem cells in the brain of rats with vascular dementia. In this study, a vascular dementia model was established by repeatedly clipping and reperfusing the bilateral common carotid arteries of rats in combination with an intraperitoneal injection of a sodium nitroprusside solution. Seven days after establishing the model, rats were intragastrically given EGb761 at 50 mg/kg per day. Learning and memory abilities were assessed using the Morris water maze and proliferation of endogenous neural stem cells in the subventricular zone and dentate gyrus were labeled by 5-bromo-2-deoxyuridine immunofluorescence in all rats at 15 days, and 1, 2, and 4 months after model establishment. The escape latencies in Morris water maze tests of rats with vascular dementia after EGb761 treatment were significantly shorter than the model group. Immunofluorescence staining showed that the number and proliferation of 5-bromo-2-deoxyuridine-positive cells in the subventricular zone and dentate gyrus of the EGb761-treated group were significantly higher than in the model group. These experimental findings suggest that EGb761 enhances proliferation of neural stem cells in the subventricular zone and dentate gyrus, and significantly improves learning and memory in rats with vascular dementia.
基金supported by the Shanghai Rising-Star Program (No. 10QH1400400)the Natural Science Foundation of Science and Technology Commission of Shanghai MunicipalityChina (No. 10JC1402302)
